CN120091817A

CN120091817A - Combination therapy of orbicetrapib and ezetimibe and fixed-dose pharmaceutical compositions

Info

Publication number: CN120091817A
Application number: CN202380070546.1A
Authority: CN
Inventors: 乔安妮·莱斯利·克雷格; 崔盛; 迈克尔·哈维·戴维森; 马克·迪特马希; J·J·P·卡斯特莱因; 瓦列里娅·尼古拉耶夫娜·斯莫伦斯基亚; 安德烈亚斯·勒内·罗特利; 岸田宗己; 克里斯托弗·J·博尔斯
Original assignee: New Amsterdam Pharmaceutical Pte Ltd
Current assignee: New Amsterdam Pharmaceutical Pte Ltd
Priority date: 2022-08-22
Filing date: 2023-08-22
Publication date: 2025-06-03
Also published as: JP2025527686A; IL318925A; KR20250065346A; US20250186442A1; EP4577207A1; MX2025001876A; CA3265606A1; AU2023328966A1

Abstract

The present disclosure relates to stable pharmaceutical compositions comprising fixed dose combinations of olanexidine and ezetimibe or salts, solvates, or derivatives of olanexidine and ezetimibe. The present disclosure further describes the use of ezetimibe and eubiperidine, e.g., in such a fixed dose combination, for the preparation of a medicament for treating a subject in need of lowering LDL-cholesterol or a subject suffering from hyperlipidemia or mixed dyslipidemia, and methods of treatment for the subject.

Description

Orbicifacipimox and ezetimibe combination therapy and fixed dose pharmaceutical composition

Technical Field

The present disclosure relates to a fixed dose pharmaceutical composition comprising obrisetrapib (obicetrapib) and ezetimibe and the use thereof for the preparation of a medicament required for a subject in need of lowering LDL cholesterol or an inpatient with heterozygous familial hypercholesterolemia (HeFH) and/or established diagnosis of atherosclerotic cardiovascular disease (ASCVD), and for the treatment of said subject or inpatient.

Background

Despite advances in therapy, cardiovascular disease (cardiovascular disease, CVD) remains a leading cause of global death, with over 1700 million deaths occurring annually. For many years, abnormal cholesterol levels have been known to be associated with increased risk of cardiovascular disease (CVD), such as cardiomyopathy, atherosclerosis, and myocardial infarction. In particular, individuals with high levels of low-density lipoprotein (low-density lipoprotein, LDL) cholesterol and very low-density lipoprotein (VLDL) cholesterol combined with low levels of high-density lipoprotein (high-density lipoprotein, HDL) cholesterol were observed to be most at risk of cardiovascular disease.

In the primary and secondary prevention of cardiovascular events, lowering low density lipoprotein cholesterol (low-density lipoprotein cholesterol, LDL-C) is the primary goal of therapy. Although statin therapy is the primary means of lowering LDL-C, a substantial percentage of patients taking the statin formulation either fail to reach the target blood lipid level for statin therapy or are partially or completely intolerant to statin. In order to reduce the risk of non-fatal or fatal cardiovascular disease recurrence, such patients are advised to use alternative lipid-lowering pharmaceutical formulations in combination.

One class of alternative therapeutic drug formulations are cholesterol absorption inhibitors (Cholesterol Absorption Inhibitor, CAI). CAI prevents small intestine ingestion of cholesterol by blocking ingestion of micellar cholesterol, thereby reducing inclusion of cholesterol esters in chylomicrons (chylomicron) and chylomicron remnants. CAI reduces the amount of cholesterol circulating back to the liver, thereby increasing hepatic LDL receptor activity and increasing the clearance of LDL cholesterol particles from the blood stream.

A known example of CAI is ezetimibe, a compound previously known as "Sch-58235" from pioneer-Plough, and marketed under brand names of beneficiary purity (Ezetrol) and Ai Zeting (Zetia) (moxadong (MERCK SHARP & Dohme)/Merck) among many commercial products. Ezetimibe has the IUPAC name (3 r, 4S) -1- (4-fluorophenyl) -3- [ (3S) -3- (4-fluorophenyl) -3-hydroxypropyl ] -4- (4-hydroxyphenyl) azetidin-2-one. Ezetimibe is often administered as a monotherapy or in an additional combination therapy. Typically, ezetimibe dosage forms are tablets comprising 10mg ezetimibe for oral administration.

Another therapeutic agent is an inhibitor of Cholesterol Ester Transfer Protein (CETP). CETP is a plasma protein secreted primarily by liver and adipose tissue. CETP mediates transfer of cholesterol esters from HDL to apolipoprotein B (ApoB) containing particles (mainly LDL and VLDL) to exchange triglycerides (TRIGLYCERIDE, TG) to reduce cholesterol levels in HDL and facilitate cholesterol levels in (V) LDL. Thus, CETP inhibition has been postulated to retain cholesterol esters in HDL-C and to reduce the cholesterol content of the atherogenic (atherogenic) ApoB moiety.

Although there is evidence supporting the potential of CETP inhibition in reducing cardiovascular morbidity, clinical development of CETP inhibitors is not easy and a variety of CETP inhibitors have been abandoned at various stages of clinical development. Olanzapine (also known as TA-8995) is currently undergoing clinical evaluation.

In the treatment of subjects suffering from hyperlipidemia or mixed dyslipidemia, there remains a need for improved therapies to reduce the risk of cardiovascular events, for example by combination therapies.

As will be explained in more detail below, the inventors of the present invention have found that significant improvements in lipid profile (blood lipid profile) are obtained with the combined treatment of obipratropium and ezetimibe, and thus, in general, aspects of the present invention provide a method of treatment comprising concomitant administration of obipratropium and ezetimibe.

Combination therapy requires co-administration of multiple pills as indicated by the precise prescription of a physician prescribing such treatment for the patient. Since each drug in combination therapy may have its own set of instructions, it is often cumbersome to have patients follow such instructions for a long time, and this is more complicated for the treatment of chronic diseases (e.g. chronic diseases requiring lipid lowering) and for the patient or the patient's caregivers. Such difficulties often lead to non-compliance with the medical prescription, thereby compromising efficacy, increasing the risk of adverse reactions, and in many cases, developing resistance or altering sensitivity of the target receptor/protein.

The preparation of fixed dose combinations of different drugs in a single pharmaceutical dosage form is often challenging due to a variety of factors such as the physicochemical incompatibility of the active pharmaceutical ingredient (active pharmaceutical ingredient, API), such as API-API interactions, excipient-excipient interactions and drug-excipient interactions. Physicochemical incompatibilities of active ingredients include challenges arising from differences in the physicochemical properties and behavior of APIs, for example, pKa, logP, solubility (solubility), hygroscopicity, photosensitivity, particle size, flowability, compressibility, melting point, or any such other parameter of one active ingredient may not be suitable for stability of another API in a formulation. Because the size and shape of the dosage form need to be controlled within the proportions of conventional administered pills, the total amount of excipients that can be used to achieve the desired stability and solubility (dissolution) of each API in a fixed dose formulation is limited, as compared to preparing a stable formulation with a single API. The incompatibility of some excipients of one or more drugs in a fixed dose combination further limits the choice of formulation scientists. A further challenge in this regard exists when one or both APIs have poor water solubility, solubility or dissolution patterns that differ (e.g., one is a soluble drug and one is an insoluble or poorly soluble drug; or one is a lipophilic drug and the other is a hydrophilic drug). In a fixed dose combination, the interaction of one drug or impurity thereof with another drug or impurity thereof may further affect the stability, efficacy, or solubility of one or both drugs.

Ezetimibe is a nearly insoluble drug and has low solubility throughout the physiological pH range. Ezetimibe is also incompatible with many commonly used excipients and presents stability problems, for example the presence of polyethylene glycol (polyethylene glycol, PEG) in the coating layer can lead to increased tetrahydropyran impurities in ezetimibe. Moreover, ezetimibe is an inherently incompressible and poorly flowing API (see e.g. EP 2168573 A1), and thus preparing a tablet formulation of ezetimibe is quite challenging.

Olanzapine also has poor water solubility in the physiological pH range and negatively affects ezetimibe solubility (unpublished data). To the applicant's knowledge, no fixed dose combination of ezetimibe with objected has been found in the art that (i) can remain stable over a long period of time without significantly increasing the level of deleterious impurities, (ii) there is no significant API-API interactions, API-excipient interactions or excipient-excipient interactions that can render such a composition unsuitable for human use, (iii) can consistently provide a desired dissolution profile for each of the two ingredients over its shelf life that is comparable to or better than a formulation with a single drug, (iv) is easy to formulate and does not pose challenges to the processability of the ingredients during formulation and scale-up (scale-up) manufacturing, (v) is capable of achieving the desired bioavailability when co-administered as two separate formulations of each drug, when orally administered by humans, and has bioequivalence with the same dose of the two active ingredients, and (vi) provides improved patient compliance that is comparable to or better than a formulation with a single drug, thereby exhibiting prolonged equivalent or poorer levels of protein and/or receptor-side effects due to such drug exposure, for example, and no prolonged periods of drug exposure to such a therapeutic drug.

Thus, there remains a need for a fixed dose combination formulation of ezetimibe and obreplacement for use in treating a subject suffering from hyperlipidemia or mixed dyslipidemia that meets all of the criteria described above and reduces the risk of cardiovascular events.

Disclosure of Invention

As already mentioned above, the inventors of the present invention have found that significant improvement of the lipid profile is obtained with the combined treatment of obipratropium and ezetimibe even in subjects who do not respond adequately to (high intensity) statin treatment, e.g. in high intensity statin (HIGH INTENSITY STATIN, HIS) low responders. More particularly, as described in the experimental part of this document, it has now been shown in phase 2b clinical trials ("ROSE 2"; NCT 05266586) that the combination of olanzapine (10 mg) and ezetimibe (10 mg) is well tolerated and reduces the median LDL-C by 59%, which clearly demonstrates the superadditive effect (supra ADDITIVE EFFECT). In particular, the median value of LDL-C in patients treated with obotrypsin was reduced by 39%, which means that the additional use of ezetimibe on the basis of obotrypsin additionally reduced LDL-C (median value) by about 32%. This is (largely) exceeded the reduction in LDL-C normally obtained with ezetimibe, which typically reduces LDL-C levels by 15% to 22% (in hyperlipidemic patients) with ezetimibe monotherapy, whereas ezetimibe typically reduces LDL-C levels incrementally by 15% to 20% when used in combination with statins (see, e.g., catapano et al European J.Heart [ European Heart Journal ] (2016) 37, 2999-3058). Significant increases in ApoB and Lp (a) levels were also demonstrated in the experiments.

Accordingly, one aspect of the present invention relates to a fixed dose pharmaceutical composition comprising obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable excipient, for example a fixed dose pharmaceutical composition wherein the composition is a two-component composition and wherein one of the components comprises ezetimibe and the other component comprises obbestrepide.

One embodiment relates to a fixed dose pharmaceutical composition comprising obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable excipient, wherein when the pharmaceutical composition is dissolved in 500ml of a solution comprising 0.45% SLS in 0.05M sodium acetate buffer having a pH of 4.5 at 37±0.5 ℃ in a USPII-type device at a rotation speed of about 75rpm, at least about 60%, preferably at least about 70%, and more preferably at least about 80%.

One embodiment relates to a fixed dose pharmaceutical composition comprising obbespride or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable excipient, wherein upon oral administration of the composition to a subject, the area under the curve (AUC 0-and/or AUC 0-t) of obbespride and/or 90% confidence interval of the geometric mean of Cmax is in the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC 0-and/or AUC 0-t) and/or Cmax, respectively, of obbespride obtained upon oral administration of a reference pharmaceutical composition (REFERENCE PHARMACEUTICAL COMPOSITION) to a similar subject, wherein the reference pharmaceutical composition comprises obbespride or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition comprises ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, alone or in the same order as the fixed dose of the further pharmaceutical composition.

Another embodiment relates to a fixed dose pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable excipient, wherein upon oral administration of the composition to a subject, ezetimibe and/or ezetimibe glucuronide has a 90% confidence interval of the geometric mean of the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or Cmax of ezetimibe and/or ezetimibe glucuronide, respectively, that is obtained when orally administering a reference pharmaceutical composition to a similar subject, respectively, the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or ezetimibe glucuronide, preferably 80% to 125%, and more preferably 90% to 110%, wherein the reference pharmaceutical composition comprises ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, at an equivalent dose, and wherein the reference pharmaceutical composition comprises ezetimibe and/or a pharmaceutically acceptable salt or co-crystal thereof, administered alone or in combination with the fixed dose or co-crystal thereof, respectively.

It has surprisingly been found that a fixed dose pharmaceutical composition of ezetimibe and obsemide can be kept stable over a long period of time without significantly increasing the level of deleterious impurities or without forming substantial amounts of new impurities. It has also surprisingly been found that a fixed dose pharmaceutical composition of ezetimibe and olanzapine does not have any significant API-API interactions, drug-excipient interactions and/or excipient-excipient interactions that may render the formulation unsuitable for use.

Even more surprisingly it was found that the pharmaceutical composition consistently provides dissolution profiles of ezetimibe and objected over its whole shelf-life period, which is equivalent to the solubility achieved by formulations comprising only a single drug. Since the stable composition provides the desired dissolution profile through a single bolus, it surprisingly overcomes problems associated with co-administration of multiple boluses of a single pharmaceutical formulation, such as poor patient compliance, suboptimal therapeutic effects, and increased risk of undesirable side effects (e.g., receptor development resistance or high sensitivity). This makes the fixed dose composition a particularly relevant treatment for chronic treatment of patients in need of lipid lowering therapy, thus making such therapy suitable.

A second aspect relates to a fixed dose pharmaceutical composition comprising obrisetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable excipient for use in reducing LDL cholesterol in a patient in need of lowering LDL cholesterol and/or raising HDL cholesterol, in a patient suffering from heterozygous familial hypercholesterolemia (HeFH) and/or in a patient diagnosed with atherosclerosis cardiovascular disease (ASCVD).

The present invention also provides a method of treating a subject in need thereof, the method comprising concomitant treatment of the subject with obipratropium or a pharmaceutically acceptable salt, solvate or co-crystal thereof and ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, preferably in the form of a fixed dose pharmaceutical composition as defined herein.

More particularly, the present invention relates to the following aspects.

One aspect of the invention relates to a method for prophylactic and/or therapeutic treatment of a subject suffering from or at risk of suffering from CVD, in particular ASCVD, comprising concomitant treatment of the subject with obrisetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof and ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Another aspect of the invention relates to a pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and/or obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof for use in a method for prophylactic and/or therapeutic treatment of a subject suffering from CVD, in particular ASCVD, or at risk of suffering from CVD, in particular ASCVD, wherein the method comprises concomitant treatment of the subject with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof and obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the pharmaceutical composition is a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a method of synergistically reducing LDL-C plasma levels in a subject in need thereof, the method comprising concomitant treatment of the subject with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof and obbezotepine or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof and/or obbesitrapi or a pharmaceutically acceptable salt, solvate or co-crystal thereof for use in a method of synergistically lowering LDL-C plasma levels in a subject in need thereof, the method comprising concomitant administration of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof and/or obbesitrapi or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a method of synergistically slowing the progression and/or progress of CVD (more particularly ASCVD), and/or synergistically reducing the risk and/or occurrence of CVD-related events (particularly ASCVD-related events), in a subject in need thereof, the method comprising concomitant administration of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof and obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

A further aspect of the invention relates to a pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and/or obbesitrapi or a pharmaceutically acceptable salt, solvate or co-crystal thereof for use in a method of synergistically slowing the development and/or progression of CVD (more particularly ASCVD) and/or synergistically reducing the risk and/or occurrence of a CVD-related event (particularly an ASCVD-related event) in a subject in need thereof, the method comprising concomitant treatment of the subject with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and obbesitrapi or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a method of enhancing, preferably synergistically enhancing, the LDL-C lowering effect of olanzapine or a pharmaceutically acceptable salt, solvate or co-crystal thereof in a subject in need thereof, the method comprising concomitant treatment of the subject with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof for use in a method of enhancing, preferably synergistically enhancing, the LDL-C lowering effect of obrisentan or a pharmaceutically acceptable salt, solvate or co-crystal thereof in a subject in need thereof, the method comprising concomitant administration of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

Yet another aspect of the invention relates to a method of enhancing, preferably synergistically enhancing, the therapeutic efficacy of olanzapine, or a pharmaceutically acceptable salt, solvate or co-crystal thereof, particularly in the treatment and/or prevention of CVD, more particularly ASCVD, in a subject in need thereof, said method comprising concomitant administration of ezetimibe, or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

A further aspect of the invention relates to a pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof for use in a method of enhancing, preferably synergistically enhancing, the therapeutic efficacy of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, in particular in the treatment and/or prevention of CVD, more particularly ASCVD, in a subject in need thereof, the method comprising concomitant administration of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof. In a preferred embodiment of the invention, the method comprises administering a fixed dose pharmaceutical composition as defined herein.

A further aspect of the invention relates to the use of obipratropium or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and/or ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, for the manufacture of a medicament for use in any one of the previously defined methods. In a preferred embodiment of the invention, the agent is a fixed dose pharmaceutical composition as defined herein.

Other aspects of the invention relate to a kit (kit) comprising a package containing a plurality of pharmaceutical unit dosage forms, including pharmaceutically acceptable salts, hydrates or solvates, such as fixed dose pharmaceutical compositions as defined herein, and a pharmaceutical instruction (leaflet) containing printed instructions for use for repeated self-administration of the unit dosage forms for the treatment and/or prevention of CVD, in particular ASCVD, by the combined use of an obbesitrate treatment and ezetimibe therapy.

It is to be understood that such aspects of the invention relate to the same composition, the same method of treatment, the same subject, and the like, unless explicitly stated otherwise.

In certain preferred embodiments of the present invention, the salt of obipratropium included in the pharmaceutical composition of the present invention, used in the method of the present invention, included in a unit dosage form (included in a pharmaceutical kit), etc., is the amorphous calcium salt of obipratropium.

The specific details and preferred embodiments of the methods described above, as well as the compositions and pharmaceutical kits used therein, will be apparent to persons skilled in the art upon reference to the following detailed description and the accompanying experimental section.

Definition of the definition

The obipratropium, also known as "TA-8995", has the following chemical name and chemical structure:

{4- [ (2- { [3, 5-bis (trifluoromethyl) benzyl ] [ 2R, 4S) -1- (ethoxycarbonyl) -2-ethyl-6- (trifluoromethyl) -1,2,3, 4-tetrahydroquinolin-4-yl ] amino } pyrimidin-5-yl) oxy ] butanoic acid }

Ezetimibe, also known as "Sch-58235," has the following chemical name and chemical structure:

(3R, 4S) -1- (4-fluorophenyl) -3- [ (3S) -3- (4-fluorophenyl) -3-hydroxypropyl ] -4- (4-hydroxyphenyl) azetidin-2-one.

Both obipratropium and ezetimibe can also be used as different salt forms, solvates or co-crystals. The obipratropium and ezetimibe may also be formulated as prodrugs.

The term "apolipoprotein" as used herein has its conventional meaning and refers to a protein that binds to a lipid to form a lipoprotein.

The term "apolipoprotein B" (ApoB) as used herein has its conventional meaning and refers to a protein encoded by the ApoB gene.

The term "pharmaceutical composition" as used herein has its conventional meaning and refers to a pharmaceutically acceptable composition.

The term "pharmaceutically acceptable" as used herein has its conventional meaning and refers to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of mammals, especially humans, without excessive toxicity, irritation, allergic response and other problem complications commensurate with a reasonable benefit/risk ratio.

The term "carrier" as used herein has its conventional meaning and refers to a pharmaceutically acceptable diluent, adjuvant, excipient or vehicle with which the pharmaceutically active ingredient is administered.

The term "excipient" as used herein has its conventional meaning and refers to a pharmaceutically acceptable ingredient commonly used in pharmaceutical technology for the preparation of granular, solid or liquid oral dosage formulations.

The term "salt" as used herein has its conventional meaning and includes both acid addition salts and base salts of pharmaceutically active compounds.

The term "solvate (solvate)" as used herein has its conventional meaning and refers to a compound formed by solvation, for example as a combination of solvent molecules and solute molecules or ions. Well known solvent molecules include water, alcohols, nitriles and polar organic solvents.

The term "subject" as used herein refers to a person suffering from or at risk of suffering from a disease or disorder. The terms "subject" and "patient" are used interchangeably herein.

The term "increased risk (INCREASED RISK)" has its conventional meaning and refers to a situation in which a subject, preferably a human subject (male or female), is at increased risk of experiencing a cardiovascular event based on its risk profile, including LDL-cholesterol levels above 70mg/dL, e.g. above 2.6mmol/l [100.54mg/dL ], compared to a subject with lower levels.

The term "treatment" as used herein has its conventional meaning and refers to curative (curative) treatment, palliative (palliative) treatment, and prophylactic (prophylactic) treatment.

The term "cardiovascular disease (cardiovascular disease)" as used herein has its conventional meaning and includes clinical manifestations of arteriosclerosis, peripheral vascular disease, angina pectoris, ischemia, cardiac ischemia, stroke, myocardial infarction, reperfusion injury, restenosis after angioplasty (restenosis after angioplasty), hypertension, cerebral infarction and cerebral stroke.

The term "cardiovascular event (cardiovascular event)" as used herein has its conventional meaning and refers to the occurrence of a myocardial infarction, stroke, coronary death (coronary death) or the necessity of conducting a coronary revascularization (coronary revascularization) (Ference, 2017).

The term "hypercholesterolemia (hypercholesterolemia)" as used herein has its conventional meaning and refers to a disease in which high levels of cholesterol are present in the blood.

The term "Hyperlipidemia (HYPERLIPIDAEMIA)" as used herein has its conventional meaning and refers to a disease in which a high content of lipids is found in blood.

The term "mixed dyslipidemia (mixed dyslipidaemia)" as used herein has its conventional meaning and refers to diseases in which LDL cholesterol and triglyceride levels in the blood are elevated with low levels of HDL cholesterol.

The term "statin intolerant (statin intolerant)" as used herein has its conventional meaning and refers to a subject being unable to tolerate two or more statins, one of which is a low dose, because of the adverse safety effects that are present that begin or increase during statin therapy and that are eliminated or ameliorated when statin is deactivated, in this regard reference is also made to a similar definition as approved by the FDA in the bevacizide (Ai Sipa to benefactor s (Esperion)) phase III trial.

The term "cholesterol absorption inhibitor" (cholesterol absorption inhibitor, CAI) as used herein has its conventional meaning and refers to a compound that reduces LDL-C by blocking intestinal and biliary absorption of cholesterol. One known cholesterol absorption inhibitor is ezetimibe.

The term "cholesteryl ester transfer protein inhibitor (cholesteryl ESTER TRANSFER protein inhibitor)" (CETP inhibitor) as used herein has its conventional meaning and refers to a class of compounds which inhibit the CETP receptor in mammals. One known CETP inhibitor is obrisetrapib.

The term "unit dosage form (unit dosage form)" has its conventional meaning and refers to a dosage form capable of effective administration to a subject, preferably a human, that can be readily handled and packaged, held as a physically and chemically stable unit dosage comprising a therapeutic pharmaceutical formulation (i.e., obrisetrapib or a combination of therapeutic pharmaceutical formulations (e.g., obrisetrapib and ezetimibe)).

The term "fixed dose combination (fixed dose combination)" as used herein has its conventional meaning and refers to a combination of two or more drugs or active ingredients in a defined dose that is present in a single dosage unit (e.g., tablet or capsule) and administered in this manner.

The term "free dose combination (free dose combination)" as used herein has its conventional meaning and refers to a combination of two drugs or active ingredients administered simultaneously but as two distinct dosage units.

The term "effective amount (EFFECTIVE AMOUNT)" or "therapeutically effective amount (therapeutically effective amount)" refers to an amount sufficient to effect treatment as defined herein when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending on the patient being treated, the weight and age of the patient, the severity of the disease condition, the manner of administration, and the like, as can be readily determined by one of ordinary skill in the art.

Unless specifically stated otherwise, when a compound may take the form of an alternative tautomer (tautomeric), a regioisomer (regioisomeric) and/or a stereoisomer (stereoisomeric), all of which are intended to be within the scope of the claimed subject matter. For example, when a compound is described as a particular optical isomer D-or L-it is intended that both optical isomers are encompassed herein. For example, when a compound is stated as having one of two tautomeric forms, both tautomers are intended to be encompassed herein. Thus, the compounds provided herein may be enantiomerically pure (enantiomerically pure), or be stereoisomers or diastereomeric mixtures (stereoisomeric or diastereomeric mixture). The compounds provided herein may contain chiral centers. Such chiral centers may be in the (R) configuration or the (S) configuration, or may be a mixture of the (R) configuration and the (S) configuration. Chiral centers of compounds provided herein can undergo epimerization in vivo (epimerization). Thus, one skilled in the art will recognize that for a compound that undergoes epimerization in vivo, administration of the compound in its (R) form is equivalent to administration of the compound in its (S) form.

The present disclosure also encompasses all suitable isotopic variations of the compounds according to the present disclosure, whether or not the isotopic variation system is radioactive. Isotopic variations of compounds according to the present disclosure are understood to mean compounds in which at least one atom in a compound according to the present disclosure has been exchanged for another atom having the same atomic number but an atomic mass different from the atomic mass usually or predominantly present in nature. Examples of isotopes that can be incorporated into compounds according to the present disclosure are isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, chlorine, bromine, and iodine, such as ² H (deuterium), ³ H (tritium )、¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、¹⁸F、³⁶Cl、⁸²Br、¹²³I、¹²⁴I、¹²⁵I、¹²⁹I, and ¹³¹ i. Particular isotopic variants of compounds according to the present disclosure, particularly those in which one or more radioisotopes have been incorporated, may be useful, for example, for examining the mechanism of action (MECHANISM OF ACTION) or the distribution of the active compound in the body. In vivo half-life extension or reduction in the amount of active agent required) to produce particular therapeutic benefits in some embodiments, hydrogen atoms of the compounds set forth herein may be replaced with deuterium atoms, in certain embodiments, unless otherwise specified, the isotopic variants of the compounds according to the present disclosure may be prepared by a variety of methods using corresponding isotopic modifications of the specific reagents and/or starting compounds therein, including, for example, the methods set forth below and in the working examples.

Thus, any of the embodiments described herein is intended to include a single stereoisomer, a mixture of stereoisomers, and/or an isotopic form of the compound.

Unless otherwise indicated, the term "about" or "approximately" means an acceptable error for a particular value determined by one of ordinary skill in the art, depending in part on how the particular value is measured or determined. In certain embodiments, the term "about" or "approximately" means within 1,2, or 3 standard deviations. In certain embodiments, the term "about" or "approximately" means within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.25%, 0.2%, 0.1%, or 0.05% of a given value or range. The term "about" means, unless stated otherwise, rounded up or down to the nearest integer within + -10% of the explicitly recited value.

Drawings

FIG. 1 cumulative undersize distribution curve for small-scale batches of a fixed dose combination composition of ezetimibe and 5mg of olanzapine

FIG. 2 retention curve of small scale batches of a fixed dose combination composition of ezetimibe and 5mg of olanzapine

FIG. 3 compares the dissolution profiles of ezetimibe at pH 6.8 by a differential dissolution method for small-scale batches of a fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 4 compares dissolution profiles of olanzapine at pH 6.8 by differential dissolution method for small scale batches of fixed dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 5 comparison of dissolution profiles of ezetimibe by differential dissolution at pH 4.5 for small-scale batches of fixed-dose combination compositions of 10mg ezetimibe and 5mg olanzapine

FIG. 6 compares the dissolution profile of olanexidine for a small scale batch of a fixed dose combination composition of 10mg ezetimibe and 5mg of olanexidine at pH 6.8 for a stress stability study of batch a4459/05/05

FIG. 7 compares the dissolution profile of olanexidine for a small scale batch of a fixed dose combination composition of 10mg ezetimibe and 5mg of olanexidine at pH 6.8 for a stress stability study of batch a4459/05/06

FIG. 8 compares the dissolution profile of olanexidine for a small scale batch of a fixed dose combination composition of 10mg ezetimibe and 5mg of olanexidine at pH 6.8 for a stress stability study of batch a4459/05/07

FIG. 9 compares the dissolution profile of olanexidine for a small scale batch of a fixed dose combination composition of 10mg ezetimibe and 5mg of olanexidine at pH 6.8 for stress stability studies for batch a4459/05/08

FIG. 10 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/05 at ph 6.8 for a small scale batch of a fixed dose combination composition of 10mg ezetimibe and 5mg obsemipiride

FIG. 11 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/06 at pH 6.8 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 12 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/07 at pH 6.8 for small-scale batches of fixed-dose combination compositions of 10mg ezetimibe and 5mg obsemipiride

FIG. 13 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/08 at pH 6.8 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 14 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/05 at pH 4.5 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 15 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/06 at pH 4.5 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg obsemipiride

FIG. 16 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/07 at pH 4.5 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 17 compares the ezetimibe dissolution profile of stress stability study for batch a4459/05/08 at pH 4.5 for small-scale batches of fixed-dose combination composition of 10mg ezetimibe and 5mg of olanzapine

FIG. 18 fixation of 10mg ezetimibe with 10mg obipratropium undersize cumulative distribution curve% for small-scale batches of composition

FIG. 19 dissolution profile of ezetimibe at a small scale of 10mg and fixed dose of olanzapine (free acid) composition at 10mg

FIG. 20 Ezetimibe dissolution profile (50 rpm) for small-scale 10mg Ezetimibe and 10mg Obiutrepidine (free acid) fixed dose composition

FIG. 21 dissolution profile (75 rpm) of ezetimibe at a small scale of 10mg ezetimibe and 10mg of an olanzapine (free acid) fixed dose composition

FIG. 22 stress stability dissolution results of the obbestrech prototype C200 BN A4459/19/03 for a small scale 10mg ezetimibe and 10mg obbestrech (free acid) fixed dose composition

FIG. 23 stress stability dissolution results of the Obipsilateral C amplified BN A4459/19/02 for a small scale 10mg ezetimibe and 10mg Obipsilateral (free acid) fixed dose composition

FIG. 24 stress stability dissolution results of ezetimibe prototype C-amplified BN A4459/19/02 for small-scale 10mg ezetimibe and 10mg obsemipiride (free acid) fixed dose compositions

FIG. 25 stress stability dissolution results of ezetimibe prototype C-amplified BN A4459/19/02 for small-scale 10mg ezetimibe and 10mg obsemipiride (free acid) fixed dose compositions

FIG. 26 cumulative undersize distribution curve for small-scale FDC1 composition

FIG. 27 dissolution profile of obiprift of FDC1 prototype

FIG. 28 Ezetimibe dissolution curve of FDC1 prototype

FIG. 29 cumulative undersize distribution curve for small-scale FDC2 composition

FIG. 30 dissolution profile of obiprifepin for small scale FDC2 compositions

FIG. 31 Ezetimibe dissolution profile for Small-Scale FDC2 composition

FIG. 32 dissolution profile of obiprifepin for small-scale FDC2 coated tablets obtained by differential method

FIG. 33 dissolution profile of obiprifepin for small-scale FDC2 coated tablets obtained by QC method

FIG. 34 Ezetimibe dissolution profile of small-scale FDC2 coated tablets obtained by QC method

Figure 35 dissolution profile of obiprift of small scale prototype 2 of FDC2 coated tablets according to stress stability

FIG. 36 shows an enlarged batch of cumulative undersize distribution curve according to Ezetimibe dissolution plot 37 for prototype 2 of FDC2 coated tablets with stress stability

FIG. 38 dissolution profile of obiprift from an enlarged batch of FDC1 particles

FIG. 39 Ezetimibe dissolution curve for FDC1 particles from an enlarged batch

FIG. 40 Ezetimibe dissolution curve for FDC2 final blend from an enlarged batch

Fig. 41 dissolution profile 42 of obsemipirtine for uncoated tablets of FDC1 enlarged lot under different compression forces, ezetimibe dissolution profile 43 of uncoated tablets of FDC1 enlarged lot under different compression forces, obsemipirtine dissolution profile 44 of uncoated tablets of FDC2 enlarged lot under different compression forces, cumulative undersize distribution profile 45 technical lot of ezetimibe dissolution profile for uncoated tablets of FDC2 enlarged lot under different compression forces

FIG. 46 dissolution profile of Orbicubip for FDC1 and FDC2 technical lots

FIG. 47 dissolution profile of Obiprifepin for FDC1 and FDC2 technical lots

FIG. 48 Particle Size Distribution (PSD) data of particles from technical lots

Fig. 49 is an X-ray powder diffraction pattern of amorphous obiprift calcium.

Fig. 50 is an X-ray powder diffraction pattern of amorphous obiprift calcium.

Fig. 51 is an X-ray powder diffraction pattern of amorphous obiprift calcium.

Fig. 52 is an infrared spectrum of amorphous obiprift calcium.

FIG. 53 is a ¹ H-NMR spectrum of amorphous obiprift calcium.

Fig. 54 is an X-ray powder diffraction pattern of crystalline obiprift calcium.

Fig. 55 is a stack of X-ray powder diffraction patterns from a stability study of crystalline obiprifflic calcium.

Fig. 56 is a stack of X-ray powder diffraction patterns according to a stability study of amorphous obiprift semi-calcium.

Fig. 57 is a polarized light micrograph of amorphous obiprift calcium.

Fig. 58 is a polarized light micrograph of crystalline obiprift calcium.

Fig. 59 is a thermogravimetric analysis of amorphous obiprift calcium.

Figure 60 is a modulated differential scanning calorimetry thermogram (with pinholes) of amorphous obiprift calcium.

Fig. 61 is a modulated differential scanning calorimetry thermogram (with pinholes) of amorphous obiprift calcium.

Figure 62 is a modulated differential scanning calorimetry thermogram (with pinholes) of crystalline obiprift calcium.

FIG. 63 is a solid-state ¹³ C-NMR spectrum of amorphous and crystalline calcium obipratropium.

FIG. 64 is a solid-state ¹³ C-NMR spectrum of crystalline obiprift calcium.

FIG. 65 is a solid-state ¹³ C-NMR spectrum of amorphous obiprift calcium.

Fig. 66 is an X-ray powder diffraction pattern of crystalline obiprift HCl and at least partially desolvated crystalline obiprift HCl.

Fig. 67 is an X-ray powder diffraction pattern of crystalline obiprift HCl.

FIG. 68 is an X-ray powder diffraction pattern of crystalline compound 1D.

FIG. 69 is a ¹ H-NMR spectrum of compound 1D.

Detailed Description

Fixed dose pharmaceutical compositions of the invention

A first aspect relates to a fixed dose pharmaceutical composition comprising obrisetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and one or more pharmaceutically acceptable excipients.

In one of the embodiments, upon oral administration of the fixed dose pharmaceutical composition to a subject, the 90% confidence interval of the geometric mean of the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or Cmax of the obbezotep is in the range of about 75% to 125%, preferably about 80% to 125%, and more preferably about 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or Cmax, respectively, of the obbezotep obtained upon oral administration of a reference pharmaceutical composition to a similar subject, wherein the reference pharmaceutical composition comprises an equivalent dose of the obbezotep or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone or in simultaneous or sequential or combined with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof in a fixed dose form.

In another embodiment, upon oral administration of the fixed pharmaceutical composition to a subject, the 90% confidence interval of the geometric mean of ezetimibe and/or ezetimibe glucuronide (AUC _0-∞ and/or AUC _0-t) is in the range of about 75% to 125%, preferably about 80% to 125%, and more preferably about 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or Cmax of ezetimibe and/or ezetimibe glucuronide, respectively, obtained upon oral administration of a reference pharmaceutical composition to a similar subject, respectively, wherein the reference pharmaceutical composition comprises an equivalent dose of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, either simultaneously or sequentially, or in a fixed dose form with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

Ezetimibe is almost insoluble in water and has poor solubility throughout the physiological pH range. For ezetimibe, achieving the required solubility and thus bioavailability under in vivo conditions is extremely challenging. This problem is further exacerbated as the dissolution rate and total amount of soluble ezetimibe is reduced by obipratropium (unpublished data). It has surprisingly been found that when the pharmaceutical composition is dissolved in 500ml of a solution in a USPII type device at 37±0.5 ℃ at a rotational speed of about 75rpm, at least about 60%, preferably at least about 70%, and more preferably at least about 80% of ezetimibe from the fixed dose pharmaceutical composition is dissolved in about 30 minutes, the solution comprising 0.45% SLS in 0.05M sodium acetate buffer having a pH of 4.5. In a preferred embodiment it has surprisingly been found that when the pharmaceutical composition is dissolved in 500ml of solution in a USPII type device at 37±0.5 ℃ at a rotational speed of about 75rpm, at least about 60%, preferably at least about 70%, and more preferably at least about 80% of ezetimibe from the fixed dose pharmaceutical composition is dissolved in about 20 minutes, said solution comprising 0.45% SLS in 0.05M sodium acetate buffer at pH 4.5.

Furthermore, it has surprisingly been found that at least about 70%, preferably at least about 80%, more preferably at least about 85%, and even more preferably at least about 90% of the obsemitrapib from the fixed dose pharmaceutical composition dissolves within about 30 minutes when the pharmaceutical composition is dissolved in 1000ml of a solution in a USPII-type device at a rotational speed of about 75rpm at 37+ -0.5 ℃, said solution comprising phosphate buffered solution at pH 6.8+0.2% w/v polysorbate 80. In a preferred embodiment it has surprisingly been found that at least about 70%, preferably at least about 80%, and more preferably at least about 85% of the obstopper from the fixed dose pharmaceutical composition dissolves in about 15 minutes when the pharmaceutical composition is dissolved in 1000ml of a solution comprising phosphate buffered solution at pH 6.8 + polysorbate 80 at 0.2% w/v in a USPII type device at 37 ± 0.5 ℃.

Ezetimibe is essentially a poorly compressible/incompressible API (see, e.g., EP 2168573 A1) and has poor flowability. Thus, it is very challenging for formulation scientists to prepare a tablet formulation of ezetimibe that not only meets the requirements of hardness, disintegration time, friability, shape and size, but also provides the desired stability and solubility. It has surprisingly been found that the composition not only meets the required specifications for solubility and stability suitable for the claimed use, but also meets the criteria for processability parameters, i.e. flowability, compressibility, disintegration time, brittleness, hardness, shape and size.

The fixed dose pharmaceutical composition may comprise a combination of 1mg to 10mg of obrisedronate and 5mg to 20mg of ezetimibe. In a preferred embodiment, the composition comprises 5mg of obrisetrapib and 10mg of ezetimibe. In a more preferred embodiment, the composition comprises 10mg of obrisetrapib and 10mg of ezetimibe.

In a preferred embodiment, the pharmaceutical composition is provided in a unit dosage form comprising 5mg of olanzapine and 10mg of ezetimibe. In a more preferred embodiment, the composition is provided in a unit dosage form comprising 10mg of olanzapine and 10mg of ezetimibe.

Wherever in the present disclosure a dose of olanzapine or ezetimibe is referred to in mg and/or relative amounts (by weight), it is meant to refer to olanzapine or ezetimibe in its free form. Whenever a salt, solvate or co-crystal of ezetimibe or obipratropium is used, for said purpose the dose shall mean a dose equivalent to the weight of ezetimibe or obipratropium, respectively, in its free form.

In certain embodiments, the pharmaceutical composition is provided in a solid oral dosage form selected from the group consisting of a caplet, a minitablet, a tablet, a granule, a bead, a pellet, a tablet, a capsule, a pill, and the like, or in a liquid oral dosage form useful in pharmaceutical preparation, including, but not limited to, a beverage, a solution, a suspension, a syrup, a drink, and an emulsion.

In one embodiment, the solid oral dosage form is provided as a two-component pharmaceutical composition. In a preferred embodiment, one component of the two-component pharmaceutical composition comprises ezetimibe and the other component comprises objected. In another preferred embodiment, only one component of the two-component pharmaceutical composition comprises both ezetimibe and obsemipirtine.

In certain embodiments, the two-component composition is a bilayer tablet formulation. In a preferred embodiment ezetimibe is present in one of the two layers of the bilayer tablet and objected is present in the other layer.

In another embodiment, the two-component system is a capsule formulation. In a preferred embodiment, the capsule may have two types of particles, one of which comprises ezetimibe and the other of which comprises obsemipiride. In a further preferred embodiment, the capsule may comprise two different types of blends or mini-tablets each comprising ezetimibe or obsemipirtine, and optionally such blends or mini-tablets may be filled in two components of the capsule that are separate from each other. In certain embodiments, each blend or minitablet is filled in a smaller capsule, or such blend is compressed into a tablet/caplet/minitablet, and then the tablet/caplet/minitablet is filled in a capsule formulation.

In another embodiment, the fixed dose pharmaceutical composition is a compressed tablet formulation comprising an extragranular component and an intragranular component. In a preferred embodiment, the intragranular component comprises ezetimibe and the extragranular component comprises obipratropium. In a more preferred embodiment, the intragranular components include both ezetimibe and obipratropium. In another embodiment, the intragranular component comprises obrisetrapib and the extragranular component comprises ezetimibe. In yet another embodiment, the extra-granular component includes both ezetimibe and objected.

The intragranular component is present in a ratio of from about 1:99 to about 99:1, preferably from about 3:97 to about 97:3, and more preferably from about 5:95 to about 95:5, to the extragranular component. In another embodiment, the intra-particulate component and the extra-particulate component are present in a ratio of about 10:90 to about 90:10, preferably about 20:80 to about 80:20 or about 30:70 to about 70:30, and even more preferably about 40:60 to about 60:40 or about 50:50.

The term "intra-particle" refers to particles that are within or present in the particles of the composition, i.e., particles that include a first set of pharmaceutically acceptable excipients (including, but not limited to, binders, disintegrants, diluents, glidants, and solvents) and optionally include one or more pharmaceutically acceptable active ingredients (ezetimibe and/or obbezotepine in this case).

The term "extra-granular" refers to the addition of a pharmaceutically acceptable component to the material after granulation, i.e., the extra-granular portion includes a second set of pharmaceutically acceptable excipients (including, but not limited to, disintegrants, diluents, lubricants, glidants, and the like). Optionally, the extra-granular component may include one or more pharmaceutically acceptable active ingredients (ezetimibe and/or obipratropium in this case).

The pharmaceutical composition may be obtained by known conventional methods such as dry granulation, wet granulation, direct compression, roll compaction (roller compaction), fluid bed granulation, rapid mixture granulation, solvent evaporation, hot melt extrusion, and the like. In a preferred embodiment, the composition is obtained by wet granulation, followed by compression of the granules into a tablet formulation or filling of such granules into capsules.

In one embodiment, the pharmaceutical composition comprises ezetimibe in the form of anhydrous ezetimibe. In another embodiment, the pharmaceutical composition comprises ezetimibe hydrate, preferably ezetimibe in the form of ezetimibe monohydrate. In yet another embodiment, the pharmaceutical composition comprises a mixture of anhydrous ezetimibe and ezetimibe water-binding compound (preferably ezetimibe monohydrate). The molar ratio of anhydrous ezetimibe to ezetimibe hydrate (preferably ezetimibe monohydrate) in the pharmaceutical composition may be in the range of 100:0 to 0:100, 99.09:0.01 to 0.01:99.09, 99.08:0.02 to 0.02:99.08, 99.07:0.03 to 0.03:99.07, 99.06:0.04 to 0.04:99.06, 99.05:0.05 to 0.05:99.05, 99.04:0.06 to 0.06:99.04, 99.03:0.07 to 0.07:99.03, 99.02:0.08 to 0.02, 99.01:0.09 to 0.09:99.01, 99:1 to 1:99, 98:2 to 2:98, 90:10 to 10:90, 70:30 to 30:70, or 50:50. In a preferred embodiment, the composition is substantially free of ezetimibe hydrate, and about 100% of ezetimibe is in the form of anhydrous ezetimibe. In another preferred embodiment, about 99.5% of ezetimibe is present as anhydrous ezetimibe, and about 0.5% of ezetimibe is present as ezetimibe water complex (preferably ezetimibe monohydrate). In a more preferred embodiment, the composition is substantially free of anhydrous ezetimibe, and about 100% of ezetimibe is in the form of ezetimibe hydrate, preferably ezetimibe monohydrate.

Ezetimibe or obsemipire or both ezetimibe and obsemipire may exist in the form of a pharmaceutically acceptable salt, solvate or co-crystal thereof. Solvates include, but are not limited to, hydrates. In addition, "salts" refer to compounds prepared by reacting an organic acid or basic drug with a pharmaceutically acceptable mineral acid or base or an organic acid or base, and as used herein, "salts" include hydrates and solvates of salts. Exemplary pharmaceutically acceptable mineral or organic acids or bases are listed in tables 1 to 8 as in Handbook of Pharmaceutical Salts [ handbook of pharmaceutically acceptable salts ] (VHCA, zurich (Zurich) 2002, pages 334-345) edited by p.h.stahl and c.g.weruth. Pharmaceutically acceptable salts of olanzapine or ezetimibe can be readily prepared by appropriately mixing a solution of such compounds with the desired acid or base. The salt may be precipitated from the solution and collected by filtration or may be recovered by evaporation of the solvent. In one embodiment, salts include, but are not limited to, hydrochloride, phosphate, sulfate, mesylate (MESYLATE SALT), ethanesulfonate (ESYLATE SALT), and besylate (besylate salt) forms. In a preferred embodiment, the composition comprises olanzapine in the form of an alkali or alkaline earth metal salt of olanzapine, preferably comprises sodium, potassium or calcium, and more preferably comprises calcium salt of olanzapine. The term "co-crystal" as used herein means a crystalline material composed of two or more distinct solids at room temperature, each containing distinctive physical characteristics (e.g., structure, melting point, and heat of fusion), but where specifically indicated there is a special case where the active pharmaceutical ingredient may be liquid at room temperature. The co-crystal may comprise a co-crystal former (co-crystal former) H-bonded to obsemitrapib and/or ezetimibe. The co-crystal former may be directly H-bonded to the active pharmaceutical ingredient or may be H-bonded to a further molecule which binds to obipratropium and/or ezetimibe. In one of the embodiments, a co-crystal may be formed between obipratropium and ezetimibe or a salt or solvate of obipratropium and ezetimibe. Solvates of active compounds that do not further include co-crystal formers are not co-crystals. The co-crystal may also be a co-crystal between the co-crystal former and a salt of ezetimibe or a salt of obsemipirtine or a salt of both ezetimibe and obsemitine. Other modes of molecular recognition may also exist, including pi stacking (pi-stacking), guest-host complexing (guest-host complexation), and van der Waals interactions (VAN DER WAALS interactions). Among the interactions listed above, hydrogen bonding is the primary interaction in forming a co-crystal, whereby non-covalent bonds are formed between one part of the hydrogen bond donor and another part of the hydrogen bond acceptor. In another embodiment, the co-crystal includes two co-crystal formations. Co-crystal formers include, but are not limited to, free acids, free bases or zwitterions, salts, inorganic base addition salts (e.g., sodium, potassium, lithium, calcium, magnesium, ammonium, aluminum) or organic base addition salts, or inorganic acid addition salts (e.g., HBr, HCl, sulfuric, nitric, or phosphoric acid addition salts), or organic acid addition salts (e.g., acetic, propionic, pyruvic, malonic, succinic, malic, maleic, fumaric, tartaric, citric, benzoic, methanesulfonic acid addition salts, ethanesulfonic acid addition salts, stearic acid addition salts, or lactic acid addition salts), anhydrous or hydrated forms (or more particularly, e.g., hemihydrate, monohydrate, dihydrate, trihydrate, tetrahydrate, pentahydrate) or salts in free form, or solvates or salts in free form. To achieve this, the ratio of active ingredient to co-crystal former may be stoichiometric or non-stoichiometric. For example, a ratio of 1:1, 1:1.5, 1:2 and 2:1 of the co-crystal former of the active ingredient (obetatrapib or ezetimibe or both obetatrapib and ezetimibe, including salts or solvates thereof) is acceptable.

In one of the embodiments, the fixed dose pharmaceutical composition comprises ezetimibe or obsemipirtine or both ezetimibe and obsemitriptan as a micronized API. The particle size distribution of this micronised API can be determined by one skilled in the art using methods well known in the art. These methods include, but are not limited to, laser diffraction (laser diffraction, LD), dynamic light scattering (DYNAMIC LIGHT SCATTERING, DLS), dynamic image analysis (DYNAMIC IMAGE ANALYSIS, DIA) or analytical analysis (SIEVE ANALYSIS). Preferably, the method employed is a laser diffraction dry powder dispersion (laser diffraction dry powder dispersion) that provides a particle size distribution by measuring the angular change in the intensity of scattered light as the laser beam passes through the dispersed particulate sample. Large particles scatter light at small angles relative to the laser beam, while small particles scatter light at large angles. The angular scattering intensity data is then analyzed to calculate particle size, which generates an undersize cumulative discrete distribution curve (cumulative undersize discrete distribution curve) that gives a volumetric particle size distribution. The particle size obtained according to this method is generally reported as volume equivalent sphere diameter (Dv). The most common percentiles recorded are Dv10, dv50, and Dv90 (also referred to as X ₁₀、X₅₀ and X ₉₀). Dv90 means that 90% by volume of the particles are smaller than a specific size and 10% by volume of the particles are larger than a specific size, dv50 means that 50% by volume of the particles are smaller than a specific size and 50% by volume of the particles are larger than a specific size, and Dv10 means that 10% by volume of the particles are smaller than this size and 90% by volume of the particles are larger than this size.

In a preferred embodiment, the composition comprises micronized ezetimibe having a Dv90 of no more than 10 μm, preferably in the range of 4 μm to 10 μm, more preferably no more than 8.5 μm, a Dv50 of no more than 4 μm, preferably in the range of about 1 μm to 4 μm, more preferably no more than 3.8 μm, and a Dv10 of no more than 1 μm.

In another preferred embodiment, the composition comprises micronized obipratropium having a Dv90 of no more than 14 μm, preferably in the range of about 5 μm to 14 μm, a Dv50 of no more than 5 μm, preferably in the range of about 3 μm to 5 μm, and a Dv10 of no more than 3 μm.

The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients. Excipients include, but are not limited to, one or more binders, surfactants, disintegrants, glidants, lubricants, diluents, chelating agents, drying agents, or absorbents. The following references, which are incorporated by reference in their entirety, disclose techniques and excipients for formulating oral dosage forms. See, the Handbook of Pharmaceutical Excipients, edited by Rowe et al, manual of pharmaceutical excipients, 9 th edition, american society of medicine (American Pharmaceuticals Association) (2020); remington, THE SCIENCE AND PRACTICE of Pharmacy, lemington, pharmaceutical science and practice, edited by Gennaro, 22 nd edition, lippincott Williams & Wilkins, liPing Kot Williams and Wilkinspir Press, 2013.

The one or more binders used in the pharmaceutical composition are preferably selected from cellulose derivatives such as methylcellulose and carboxymethylcellulose, hydroxypropyl methylcellulose, hydroxypropyl cellulose and hydroxyethyl cellulose, gelatin, dextrose, sucrose, lactose, dextrose, xylitol, sorbitol, maltitol, polymethacrylates, copolymers of polyvinylpyrrolidone and polyvinylpyrrolidone, starch pastes, pregelatinized starches, tragacanth, salts of alginic acid and alginic acid such as sodium alginate, magnesium aluminum silicate, polyethylene glycols, guar gum, bentonite. In a preferred embodiment, the binder is polyvinylpyrrolidone or a copolymer of polyvinylpyrrolidone. In a more preferred embodiment, the binder is copovidone. In an even more preferred embodiment, the adhesive is Kollidon (Kollidon) 30.

The binder may generally be present in one embodiment in an amount of from about 0.2% to about 10%, from about 0.5% to about 5%, from about 0.5% to about 2%, or from about 0.5% to about 1%, preferably from about 1.0±0.5% by weight of the particulate composition and in another embodiment by weight of the total tablet.

The one or more surfactants used in the composition are preferably surfactants having an HLB value selected from at least about 15, at least about 20, at least about 30, or at least about 40. One or more such surfactants are selected from lauric acid or salts of lauric acid, palmitic acid or salts of palmitic acid, stearic acid or salts of stearic acid and oleic acid or salts of oleic acid, polyethylene glycol glycerides, polyoxyethylene monoesters, polyoxyethylene ethylene monostearate, polyoxyethylene monolaurate, polyoxyethylene sorbitan monooleate, polyethoxylated castor oil, polyethylene glycols having a molecular weight in the range of about 2000 to 10000, propylene glycol octanoate, glycerol oleate and octanoate, esters of glycerol and fatty acids. In preferred embodiments, the one or more surfactants are selected from the group consisting of dioctyl sodium sulfosuccinate, capmul PG-8, capryol 90, capmul MCM, polysorbate 20, polysorbate 40 or polysorbate 80, or sodium lauryl sulfate. In a more preferred embodiment, the surfactant is sodium lauryl sulfate, e.g., kolliphor SLS.

The surfactant may generally be present in one embodiment in an amount of from about 0.2% to 10%, from about 0.5% to about 5%, from about 0.5% to about 2%, or from about 0.5% to about 1%, preferably from about 1.0±0.5% by weight of the total tablet, based on the weight of the particulate composition and in another embodiment.

In one of the embodiments, the composition includes a binder to surfactant ratio in the range of about 0.05:5.0 to about 5.0:0.05, preferably about 0.5:4.5 to about 4.5:0.5, more preferably about 1:4 to about 4:1, even more preferably about 1:2 to about 2:1, and most preferably about 1:1. Such ratio of binder to surfactant may be used in a particulate composition (e.g., an intra-particulate composition or an extra-particulate composition) or in a total composition for a tablet.

The pharmaceutical composition typically further comprises one or more disintegrants selected from the group consisting of crospovidone, croscarmellose sodium, carboxymethylcellulose calcium, low substituted hydroxypropylcellulose, alginic acid, sodium alginate, microcrystalline cellulose, sodium starch glycolate, and pregelatinized starch. In a preferred embodiment, the disintegrant is croscarmellose sodium or sodium starch glycolate. In a more preferred embodiment, the disintegrant is sodium starch glycolate.

The disintegrant may be present in one embodiment in an amount of from about 0.5% to about 10%, from about 1% to about 8%, from about 2% to about 5%, preferably from 2% to about 3%, from about 4% to about 5%, or from about 7% to about 8% by weight of the total tablet, based on the weight of the particulate composition and in another embodiment.

The one or more diluents used in the pharmaceutical composition are preferably selected from the group consisting of inorganic phosphates, such as calcium hydrogen phosphate, or sugar analogues and derivatives thereof, in particular lactose (e.g. lactose monohydrate or lactose anhydrous), dextrose, sorbitol, mannitol, sucrose, maltodextrin, isomaltose, or celluloses, such as microcrystalline cellulose or powdered cellulose, etc. In a preferred embodiment, the diluent is selected from lactose (e.g., lactose monohydrate), microcrystalline cellulose, and mannitol, or mixtures thereof. In a more preferred embodiment, the intragranular component comprises microcrystalline cellulose and lactose monohydrate as diluents. In another preferred embodiment, microcrystalline cellulose and mannitol are present as diluents in the extra-granular component. The diluent may be present in one embodiment in an amount of from about 10% to about 95%, preferably from about 40% to about 90%, more preferably from about 60% to about 85%, even more preferably from about 70% to about 85% by weight of the particulate composition and in another embodiment by weight of the total tablet.

The pharmaceutical composition may optionally be film coated using techniques well known in the art, such as spraying or dip coating in a conventional coating machine (coating pan) or a fluid bed processor. Alternatively, the coating may also be performed using a hot melt technique. The film coating includes a film forming polymer, one or more pharmaceutically acceptable excipients, and a pharmaceutically acceptable solvent. Examples of film formers include, but are not limited to, cellulose derivatives such as methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxymethyl ethyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose, and ethyl cellulose, polyvinyl alcohol, waxes, fatty substances, or mixtures thereof. Alternatively, a variety of brands (e.g.) Commercially available coating compositions of film-forming polymers that are put into the market are coated.

Examples of solvents for preparing the coating solution are selected from methanol, ethanol, isopropanol, n-butanol, acetone, acetonitrile, chloroform, dichloromethane, water or mixtures thereof. In a preferred embodiment, the film coating is a primary alcohol-free coating. Preferably, the primary alcohol-free coating is a coating made using water.

The glidants present in the pharmaceutical dosage form are preferably selected from the group consisting of silicon dioxide, talc, magnesium stearate, and the like. The preferred glidant is silicon dioxide (e.g.,) Or magnesium stearate (e.g., LIGAMED MF V) or a mixture thereof. Glidants may generally be present in one embodiment in an amount of about 0.1% to 10%, about 0% to about 5%, or about 1% to about 2% by weight of the particulate composition and in another embodiment by weight of the total tablet.

The lubricant present in the pharmaceutical composition is preferably selected from fatty acids or fatty acid derivatives, such as alkali metal and alkaline earth metal salts of stearic acid, lauric acid and/or palmitic acid and the like. The preferred lubricant is magnesium stearate and may generally be present in one embodiment in an amount of from about 0.1% to 10%, from about 0% to about 5%, or from about 1% to about 2% by weight of the particulate composition and in another embodiment by weight of the total tablet.

Stability is an essential quality attribute of pharmaceutical formulations that determines the shelf-life of a composition during which the composition is suitable for its intended use, both from a therapeutic and safety point of view. Stability of a pharmaceutical composition of a stable pharmaceutical composition this term means that one or more parameters governing the physical and chemical integrity of the Active Pharmaceutical Ingredient (API) remain within pharmaceutically acceptable standards during the shelf life of the product. Typically, one or more such parameters are selected from the group consisting of identification of the active ingredient in the composition by methods such as HPLC and/or Ultraviolet (UV) spectroscopy, visual appearance of the composition, measured percentage of active ingredient in the composition (ASSAY PERCENTAGE), individual and/or total percentages of related substances and/or impurities in the composition, content uniformity of the composition relative to the active ingredient, dissolution rate, microbiological limit, and the like.

Pharmaceutical compositions often lose their efficacy and/or safety over time due to loss or degradation of the active ingredient or conversion of the active ingredient to impurities commonly referred to as related substances. A stable fixed dose pharmaceutical composition will retain at least up to about 90% (w/w) ezetimibe and the claimed efficacy of obsemipine.

Ezetimibe is known to cause stability problems associated with its formulation due to interactions with excipients and/or combination drug partners. It has surprisingly been found that a fixed dose pharmaceutical composition effectively controls the levels of individual related substances and total related substances of ezetimibe during the preparation and storage of the fixed dose composition. In embodiments, the stable fixed dose pharmaceutical composition has no more than about 5% (w/w), preferably no more than about 2% (w/w), more preferably no more than about 1% (w/w), and even more preferably no more than about 0.2% (w/w) of the individual related substances of ezetimibe, and no more than about 5% (w/w), preferably no more than about 2% (w/w), more preferably no more than about 1% (w/w), and even more preferably no more than about 0.5% (w/w) of the total related substances of ezetimibe. In a preferred embodiment, the fixed dose pharmaceutical composition comprises ezetimibe and olanzapine, wherein the ezetimibe tetrahydropyran analog impurity is no more than about 2% (w/w), preferably no more than about 0.5% (w/w), more preferably no more than about 0.3% (w/w), even more preferably no more than about 0.2% (w/w).

In another embodiment, the stable fixed dose pharmaceutical composition has no more than about 5% (w/w), preferably no more than about 2% (w/w), more preferably no more than about 0.5% (w/w), even more preferably no more than about 0.3% (w/w), and most preferably no more than about 0.2% (w/w) of any unspecified individual obickie related substance, and no more than about 5% (w/w), preferably no more than about 2% (w/w), more preferably no more than about 1% (w/w), and even more preferably no more than about 0.5% (w/w) of the total related substance of obickie.

It has surprisingly been found that the pharmaceutical composition remains stable under stability conditions of a temperature of 40 ℃ and a relative humidity of 75% for at least up to 1 month, preferably for at least up to 3 months, more preferably for at least up to 6 months. In a preferred embodiment, the composition remains stable for at least up to 3 months, preferably at least up to 6 months, under stability conditions of 40 ℃ temperature and 75% relative humidity. In another preferred embodiment, the composition remains stable for at least up to 3 months, 6 months or 12 months under stability conditions of 25 ℃ temperature and 60% relative humidity. In a further preferred embodiment, the composition remains stable at room temperature for at least up to 6 months, 12 months, 18 months or 24 months.

In a preferred embodiment, the pharmaceutical composition is a tablet formulation comprising or consisting of:

a. An extragranular component comprising:

i. calcium obiprift equivalent to 10mg obiprift free acid;

Anhydrous ezetimibe or ezetimibe equivalent to 10mg ezetimibe Ezetimibe cloth Ezetimibe (R) wheat a mixture of cloth monohydrate;

A 1:1 ratio of binder to surfactant, preferably the binder and surfactant each comprise about 1.+ -. 0.5% w/w of the particles of the intra-particle component, more preferably the binder is 1.+ -. 0.5% w/w povidone or polyvinylpyrrolidone and the surfactant is 1.+ -. 0.5% w/w sodium lauryl sulfate;

preferably the disintegrant is about 2% to 8% w/w of the particles of the intra-particle component, preferably 3% to 6% w/w, more preferably about 4.5.+ -. 0.5% w/w;

one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably lactose anhydrous or lactose monohydrate, even more preferably lactose monohydrate, polysaccharides, preferably cellulose, more preferably microcrystalline cellulose, sugar alcohols, preferably sorbitol, xylitol or mannitol;

b. an extragranular component comprising:

i. a disintegrant selected from croscarmellose sodium, pregelatinized starch, or sodium starch glycolate, more preferably sodium starch glycolate, even more preferably about 4% w/w to 6% w/w sodium starch glycolate;

Optionally a lubricant, preferably magnesium stearate, more preferably about 1% w/w to 2% w/w magnesium stearate;

optionally a glidant, preferably colloidal silicon dioxide or talc or both colloidal silicon dioxide and talc, more preferably about 1% w/w to 2% w/w colloidal silicon dioxide or talc or both colloidal silicon dioxide and talc;

Optionally one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably lactose anhydrous or lactose monohydrate, even more preferably lactose monohydrate, polysaccharides, preferably cellulose, more preferably microcrystalline cellulose, sugar alcohols, preferably sorbitol, xylitol or mannitol, more preferably mannitol and microcrystalline cellulose, even more preferably about 20% w/w to about 50% w/w microcrystalline cellulose and about 1% to about 20% mannitol.

C. optionally, the pharmaceutical composition comprises a film coating, preferably the film coating is free of primary alcohols, more preferably the film coating is free of polyethylene glycols.

In another preferred embodiment, the pharmaceutical composition comprises a tablet formulation comprising or consisting of:

a. An extragranular component comprising:

i. anhydrous ezetimibe or a mixture of anhydrous ezetimibe and ezetimibe water-distribution compound equivalent to 10mg ezetimibe;

preferably the disintegrant is about 2% w/w to 8% w/w of the particles of the intra-particle component, preferably 3% w/w to 6% w/w, more preferably about 4.5.+ -. 0.5% w/w;

b. an extragranular component comprising:

i. calcium obiprift equivalent to 10mg obiprift free acid;

A disintegrant selected from microcrystalline cellulose, pregelatinized starch, or sodium starch glycolate, more preferably sodium starch glycolate, even more preferably about 4% w/w to 6% w/w sodium starch glycolate;

optionally a lubricant, preferably magnesium stearate, more preferably about 1% w/w magnesium stearate;

In a further preferred embodiment, the pharmaceutical composition is a tablet formulation comprising or consisting of:

a. An extragranular component comprising:

i. calcium obiprift equivalent to 10mg obiprift free acid;

b. an extragranular component comprising:

Disintegrating agent selected from croscarmellose sodium, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate, even more preferably about 4% w/w to 6% w/w sodium starch glycolate

Optionally a lubricant, preferably magnesium stearate, more preferably about 1% to 2% w/w magnesium stearate

Optionally a glidant, preferably colloidal silicon dioxide or talc or both colloidal silicon dioxide and talc, more preferably about 1% to 2% colloidal silicon dioxide or talc or both colloidal silicon dioxide and talc;

Another aspect relates to a pharmaceutical composition comprising obipratropium and ezetimibe or obipratropium and pharmaceutically acceptable salts, solvates or co-crystals thereof, and a pharmaceutically acceptable carrier for use in treating a subject in need of additional lowering of low density lipoprotein cholesterol as an adjunct to diet and/or maximum tolerability lipid lowering therapy for treating an adult human suffering from heterozygous familial hypercholesterolemia (HeFH) or diagnosed with atherosclerotic Cardiovascular (CV) disease (ASCVD).

A second aspect relates to the use of a fixed dose pharmaceutical composition comprising obbestrepide or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and one or more pharmaceutically acceptable excipients in the manufacture of a medicament for the treatment of a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol.

In one of the embodiments, the subject is suffering from or has hyperlipidemia or mixed dyslipidemia, heterozygous familial hypercholesterolemia (HeFH), or is diagnosed with atherosclerotic cardiovascular disease (ASCVD).

In one embodiment, the subject is partially or completely intolerant to statin drugs.

In one embodiment, the use of the pharmaceutical composition is for the treatment of a subject in need of additional lowering of low density lipoprotein cholesterol as an adjunct to diet and/or maximum tolerability lipid-lowering therapy for the treatment of adults suffering from heterozygous familial hypercholesterolemia (HeFH) or diagnosed with atherosclerotic Cardiovascular (CV) disease (ASCVD).

A third aspect relates to a method of treating a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, wherein the method comprises administering to the subject a therapeutically effective dose of a fixed dose pharmaceutical composition comprising obbesitrapi or a pharmaceutically acceptable salt, solvate or co-crystal thereof, ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and one or more pharmaceutically acceptable excipients.

In one of the embodiments, the method is for treating a subject suffering from or having hyperlipidemia or mixed dyslipidemia, heterozygous familial hypercholesterolemia (HeFH), or diagnosed with atherosclerotic cardiovascular disease (ASCVD).

In one embodiment, the subject is in need of additional lowering of low density lipoprotein cholesterol as an adjunct to diet and/or maximal tolerogenic lipid-lowering therapy for treating adults with heterozygous familial hypercholesterolemia (HeFH) or established atherosclerotic Cardiovascular (CV) disease (ASCVD).

A fourth aspect relates to a fixed dose combination pharmaceutical composition of obipratropium and ezetimibe, wherein said pharmaceutical composition is considered suitable for said use according to the second aspect or said method of treatment according to the third aspect in the following cases:

a. the fixed dose pharmaceutical composition is orally administered to a subject;

b. Measuring the concentration of olanzapine in the blood of said subject at one or more time points after administration to provide a set of olanzapine concentration/time data points to provide an area under the curve (AUC), and

C. The 90% confidence interval of the geometric mean of the area under the curve (AUC 0- ≡and/or AUC 0-t) and/or Cmax of the obbezotezole is in the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC 0- ≡and/or AUC 0-t) and/or Cmax, respectively, of the obbezotezole obtained when orally administering a reference pharmaceutical composition to a similar subject, wherein the reference pharmaceutical composition comprises an equivalent dose of obbezotezole or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone or co-administered simultaneously or sequentially with another pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, or in a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

A fifth aspect relates to a fixed dose combination pharmaceutical composition of obipratropium and ezetimibe, wherein said pharmaceutical composition is considered suitable for said use according to the second aspect or said method of treatment according to the third aspect in the following cases:

a. The fixed dose pharmaceutical composition is orally administered to a subject, ezetimibe and/or ezetimibe glucuronide in the blood of the subject is measured at one or more time points after administration to provide a set of ezetimibe and/or ezetimibe glucuronide concentration/time data points, respectively, to provide an area under the curve (AUC) of ezetimibe and/or ezetimibe glucuronide, respectively, and

B. The 90% confidence interval of the geometric mean of ezetimibe and/or ezetimibe glucuronide (AUC _0-∞ and/or AUC _0-t) and/or the 90% confidence interval of the geometric mean of ezetimibe and/or ezetimibe glucuronide, respectively, obtained when a reference pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof is orally administered to a similar subject, respectively, is in the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t) and/or the Cmax of ezetimibe glucuronide, respectively, wherein the reference pharmaceutical composition comprises an equivalent dose of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, either simultaneously or sequentially co-administered, or in a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

In one embodiment of the use according to the above aspect, t of AUC 0-t is selected from 48 hours (AUC 0-48), 72 hours (AUC 0-72), 96 hours (AUC 0-96), 144 hours (AUC 0-144), 192 hours (AUC 0-192), 240 hours (AUC 0-240), 336 hours (AUC 0-336) or AUC 0- ≡, preferably 48 hours (AUC 0-48), and more preferably 72 hours (AUC 0-72) or AUC 0-fact.

In one embodiment, the subject is a healthy human subject, preferably an adult male or female without tobacco, without nicotine, more preferably 18 to 65 years old, and optionally the human has a body mass index of 18.5 to 29.9Kg/m ².

In another embodiment, the subject is a human in need of lowering LDL cholesterol and/or raising HDL cholesterol. In preferred embodiments, the human is suffering from or has hyperlipidemia or mixed dyslipidemia, heterozygous familial hypercholesterolemia (HeFH) or is diagnosed with atherosclerotic cardiovascular disease (ASCVD).

In one embodiment, the human is partially or completely intolerant to statin drugs.

Preferably, the LDL-cholesterol level of a human subject is ≡70mg/dL, and optionally the human is not adequately controlled by its current lipid-regulating therapy.

For use of a pharmaceutical composition according to other aspects or a method of treatment, the composition may be administered to a subject in need thereof to deliver a total daily oral dose of 5mg of obbestrapib and 10mg of ezetimibe, 10mg of obbestrapib and 10mg of ezetimibe, or 20mg of obbestrapib and 20mg of ezetimibe, preferably to deliver a daily oral dose of 10mg of obbestrapib and 10mg of ezetimibe.

It has surprisingly been found that compared to the commercially available ezetimibe preparation discussed in detail in the examples sectionThe dissolution profile of ezetimibe from the fixed dose combination was found to be not poor or sometimes even better. It has also surprisingly been found that the fixed dose composition disclosed herein is bioequivalent to a combination of monotherapy drugs co-administered to a human subject. Confidence intervals (90%) of geometric mean ratios of the geometric mean ratios AUC 0-t, AUC 0- ≡and Cmax of the obbezotezole, ezetimibe and ezetimibe glucuronide from the two representative compositions FDC1 and FDC2 were found to be in the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the geometric mean ratios AUC 0-infinity and the geometric mean ratios of ezetimibe and ezetimibe glucuronide, respectively, of the obbezotezole, ezetimibe and ezetimibe glucuronide obtained from co-administration of single pharmaceutical formulations having the same doses of ezetimibe and oble, respectively, as will be discussed in detail in the experimental section below.

The fixed dose combination pharmaceutical composition of obrisetrapib and ezetimibe will be further described below by way of non-limiting example.

Methods of treatment of the invention

As described in the previous section herein, the present invention provides methods of curative and/or prophylactic treatment of a subject in need thereof. More particularly, the present invention provides methods of treating and/or preventing cardiovascular disease, particularly atherosclerotic cardiovascular disease, in such subjects using the compositions defined herein. The invention further provides methods of treating and/or preventing one or more symptoms associated with (atherosclerotic) cardiovascular disease in such a subject using the compositions defined herein. The present invention further provides methods of treating and/or preventing one or more conditions associated with and/or caused by (atherosclerotic) cardiovascular disease in such a subject using the compositions defined herein. The invention further provides methods of treating and/or preventing one or more causative factors associated with (atherosclerotic) cardiovascular disease (e.g., elevated LDL-C levels and/or elevated ApoB levels) in such subjects using the compositions defined herein. The present invention further provides methods of reducing and/or improving resistance or hyporesponsiveness of such subjects to statin therapy, particularly high-intensity statin therapy, using the compositions defined herein.

The term "treatment" used in connection with a particular disease or condition (e.g., "method of treating a disease..+ -.)" refers to curing, alleviating or eliminating the disease and/or accompanying symptoms, alleviating the extent of the disease, stabilizing (i.e., not worsening) the state of the disease, delaying or slowing the progression of the disease, ameliorating the state of the disease, prolonging survival (as compared to the expected survival when untreated), and the like. The term "preventing" (prevent, preventing or prevention) as used herein refers to reducing the risk of a subject suffering from a disease and/or developing concomitant symptoms, delaying the time to a disease in a subject, and the like. The term "treatment" as used with respect to a patient or subject (e.g., "method of treating a subject") generally refers to the act of administering a therapeutic compound to the patient or subject for any therapeutic and/or prophylactic purpose.

The term "cardiovascular disease" as used herein has its conventional meaning and refers to a disease or disorder in which the function of the cardiovascular system of a subject is impaired. Examples of cardiovascular diseases include thromboembolic diseases (e.g., arterial cardiovascular thromboembolic diseases, venous cardiovascular thromboembolic diseases, or thromboembolic diseases in the ventricles), atherosclerosis, hypertensive heart diseases, coronary artery diseases, carotid artery diseases, strokes, peripheral arterial diseases involving atherosclerosis, restenosis, arteritis, myocarditis, cardiovascular inflammation, vascular inflammation, coronary heart diseases (coronary HEART DISEASE, CHD), unstable angina (unstable angina, UA), unstable refractory angina, stable angina (stable angina, SA), chronic stable angina, acute coronary syndrome (acute coronary syndrome, ACS), myocardial infarction (first or recurrent), acute myocardial infarction (acute myocardial infarction, AMI), myocardial infarction, ischemic heart diseases, cardiac ischemia, ischemic sudden death, transient ischemic attacks, stroke, peripheral occlusive arterial diseases, venous thrombosis, deep venous thrombosis, thrombophlebitis, arterial embolism, thrombosis, cerebral thrombosis, pulmonary embolism, renal embolism, etc.

The term "atherosclerotic cardiovascular disease" as used herein refers to a particular subset of cardiovascular diseases that includes atherosclerosis as a component or precursor to a particular type of cardiovascular disease. Atherosclerosis is a chronic inflammatory reaction that occurs in the arterial vessel wall associated with retained LDL-C. It involves the formation of atheromatous plaques that can lead to narrowing of the artery ("stenosis") and can ultimately lead to partial or complete closure of the arterial opening and/or plaque rupture. Thus, atherosclerotic diseases or conditions include the consequences of atheromatous plaque formation and rupture, including, but not limited to, arterial stenosis or narrowing, heart failure, aneurysms including aortic aneurysms, aortic dissection (aortic dissection), and ischemic events (e.g., myocardial infarction and stroke).

In a particularly preferred embodiment, the atherosclerotic cardiovascular disease and/or pathology associated with an atherosclerotic cardiovascular disease which may be advantageously treated and/or prevented according to the invention is selected from the group consisting of arteriosclerosis, peripheral vascular disease, hyperlipidemia, mixed dyslipidemia betalipoproteinemia, hypoalphalipoproteinemia (hypoalphalipoproteinemia), hypercholesterolemia, hypertriglyceridemia, familial hypercholesterolemia, angina, ischemia, cardiac ischemia, stroke, myocardial infarction, reperfusion injury, restenosis after angioplasty, hypertension, cerebral infarction and cerebral stroke.

As will be apparent from the teachings of the present invention, the methods of the present invention are effective and/or aimed at reducing and/or normalizing LDL-C plasma levels. More particularly, the method is effective and/or aimed at reducing LDL-C plasma levels by at least 5% from a baseline, wherein baseline is defined as the onset of treatment with obrisetrapib and ezetimibe, more preferably by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45% or at least 50%. In further embodiments, the method is effective and/or aimed at lowering LDL-C plasma levels by at least 5mg/dL from baseline, where baseline is defined as the onset of treatment with obiprifepin and ezetimibe, more preferably by at least 10mg/dL, at least 15mg/dL, at least 20mg/dL, at least 25mg/dL, at least 30mg/dL, at least 35mg/dL, or at least 40mg/dL. In further embodiments, the method is effective and/or aimed at reducing LDL-C plasma levels to a level below 85mg/dL, preferably below 80mg/dL, below 75mg/dL, below 70mg/dL, below 65mg/dL, below 60mg/dL, below 55mg/dL or below 50 mg/dL.

As will be apparent from the teachings of the present invention, administration of ezetimibe (or a pharmaceutically acceptable salt, solvate or co-crystal thereof) in addition to obsemipirtine (or a pharmaceutically acceptable salt, solvate or co-crystal thereof) significantly enhances LDL-C reduction, particularly superadditive or synergistically enhances LDL-C reduction. More particularly, to enhance the LDL-C lowering effect of olanzapine as defined herein, the method of administering ezetimibe (or a pharmaceutically acceptable salt, solvate or co-crystal thereof) of the present invention is effective and/or aimed at further lowering LDL-C plasma levels by at least 20%, more preferably by at least 22.5%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29% or at least 30% compared to methods based on therapies using olanzapine or a pharmaceutically acceptable salt, solvate or co-crystal thereof alone (or at least without ezetimibe). In further embodiments, these methods are effective and/or aim to further reduce LDL-C plasma levels by at least 20mg/dL, more preferably by at least 22.5mg/dL, at least 25mg/dL, at least 27.5mg/dL, at least 30mg/dL, at least 32.5mg/dL, or at least 35mg/dL, as compared to methods based on therapies using obetatrapib or pharmaceutically acceptable salts, solvates, or co-crystals thereof alone (or at least without ezetimibe).

In a preferred embodiment of the invention, the method is effective and/or aimed at reducing and/or normalizing ApoB plasma levels. More particularly, the method is effective and/or aimed at reducing ApoB plasma levels by at least 5% from a baseline, wherein baseline is defined as the onset of treatment with olanexidine and ezetimibe, more preferably by at least 10%, at least 15%, at least 20%, at least 22.5%, at least 25% or at least 27.5%. In further embodiments, the method is effective and/or aimed at reducing ApoB plasma levels by at least 5mg/dL from baseline, where baseline is defined as the onset of treatment with obiprifepin and ezetimibe, more preferably by at least 5mg/dL, at least 10mg/dL, at least 15mg/dL, at least 20mg/dL, at least 22.5mg/dL, at least 25mg/dL, or at least 27.5mg/dL. In further embodiments, the method is effective and/or aimed at reducing ApoB plasma levels to a level below 80mg/dL, preferably below 75mg/dL, below 70mg/dL, below 65mg/dL, below 60mg/dL, below 57.5mg/dL or below 55 mg/dL.

In a preferred embodiment of the invention, the method is effective and/or aimed at reducing and/or normalizing Lp (a) plasma levels. More particularly, the method is effective and/or aimed at reducing Lp (a) plasma levels by at least 5% from a baseline, wherein baseline is defined as the onset of treatment with obrisedronate and ezetimibe, more preferably by at least 7.5%, at least 10%, at least 12.5%, at least 15%, at least 17.5% or at least 20%. In further embodiments, the method is effective and/or aimed at reducing Lp (a) plasma levels by at least 5nmol/L from a baseline, wherein baseline is defined as the onset of treatment with olanzapine and ezetimibe, more preferably by at least 10nmol/L, at least 15nmol/L, at least 20nmol/L, at least 25nmol/L, at least 30nmol/L, at least 35nmol/L, or at least 40nmol/L. In further embodiments, the method is effective and/or aimed at reducing Lp (a) plasma levels to a level of less than 110nmol/L, preferably less than 105nmol/L, less than 100nmol/L, less than 95nmol/L, less than 90nmol/L, less than 85nmol/L, or less than 80 nmol/L.

In some embodiments of the invention, the methods are effective and/or aimed at reducing and/or improving resistance or low responsiveness to statin therapy, in particular high intensity statin therapy. High-intensity statin therapy is a term traditionally used in the art to indicate a regimen based on the highest allowable dose of statin with the highest efficacy in lowering LDL-C, especially one that typically shows a reduction of LDL-C of ≡50% in normally responsive subjects. Of the statins currently used in clinical practice, only 20mg (daily) or 40mg (daily) of rosuvastatin and 40mg (daily) or 80mg (daily) of atorvastatin meet the criteria. In the context of the present invention, low responsiveness to HIS therapy means that the subject receiving HIS therapy fails to reach an LDL-C reduction of 35%, preferably it means that the subject receiving HIS therapy fails to reach an LDL-C reduction of 30%, an LDL-C reduction of 25%, an LDL-C reduction of 20%, an LDL-C reduction of 15% or an LDL-C reduction of 10%. Reducing and/or improving low responsiveness to HIS therapy means reducing the difference between the subject's response (LDL-C lowering) and the (average) response of the normally responding subject. In a further embodiment of the invention, the method is effective and/or aimed at normalizing the response to statin therapy.

As previously described herein, the methods of the invention are directed to treating and/or preventing a subject suffering from or at risk of suffering from CVD, particularly ASCVD.

The term "subject" refers to a living organism, typically a mammalian, particularly a human subject, suffering from or susceptible to a disease or disorder treatable using the compositions provided herein.

In a particularly preferred embodiment of the invention, the subject is a subject diagnosed with CVD, in particular ASCVD.

In further preferred embodiments of the invention, the subject is a subject considered to be at risk of having CVD, in particular ASCVD (typically having a risk above average), which may be judged, for example, by a healthcare professional.

In a preferred embodiment of the invention, the subject is a subject suffering from one or more conditions known to have causal and/or epidemiological relevance to the occurrence of (AS) CVD, such AS diabetes, hypertension, hypercholesterolemia, including overweight/obesity, metabolic syndrome, and the like. In still other preferred embodiments of the invention, the subject is a genetically predisposed subject to (AS) CVD. In still other preferred embodiments of the invention, the subject is a subject susceptible to (AS) CVD due to lifestyle/habitual factors (e.g., unhealthy diet, lack of exercise, drinking, smoking).

According to a preferred embodiment of the invention, the subject to be treated has an elevated LDL-C plasma level, typically an LDL-C plasma level of at least 70mg/dL, more preferably at least 75mg/dL, at least 80mg/dL, at least 85mg/dL, at least 90mg/dL, at least 95mg/dL or at least 100mg/dL. Furthermore, according to a preferred embodiment of the invention, the LDL-C plasma level of the subject is at least 125%, such as at least 150%, at least 175% or at least 200% of the average LDL-C plasma level of a healthy subject. Normal LDL-C (reference) values generally depend on gender and age.

According to a preferred embodiment of the invention, the subject to be treated has an elevated ApoB plasma level, typically an ApoB plasma level of at least 70mg/dL, more preferably at least 75mg/dL, at least 80mg/dL, at least 85mg/dL, at least 90mg/dL, at least 95mg/dL or at least 100mg/dL. Furthermore, according to a preferred embodiment of the invention, the ApoB plasma level of the subject is at least 125%, such as at least 150%, at least 175% or at least 200% of the average ApoB plasma level of a healthy subject. Normal ApoB (reference) values generally depend on gender and age.

According to a preferred embodiment of the invention, the subject to be treated has an elevated non-HDL-C plasma level, typically at least 100mg/dL, more preferably at least 105mg/dL, at least 110mg/dL, at least 115mg/dL, at least 120mg/dL, at least 125mg/dL or at least 130mg/dL. Furthermore, according to a preferred embodiment of the invention, the non-HDL-C plasma level of the subject is at least 125%, such as at least 150%, at least 175% or at least 200% of the average non-HDL-C plasma level of a healthy subject. Normal non-HDL-C (reference) values generally depend on gender and age.

In one embodiment of the invention, the subject is a human male. In another embodiment of the invention, the subject is a human female.

In still other preferred embodiments of the invention, the subject is at increased risk based on age (e.g., the subject is over 35 years old, over 40 years old, over 45 years old, over 50 years old, over 55 years old, over 60 years old, over 65 years old, or over 70 years old), typically in combination with one or more other risk factors defined herein.

According to certain embodiments of the invention, the subject to be treated exhibits low responsiveness to statin therapy, in particular HIS therapy. High-intensity statin therapy is a term traditionally used in the art to indicate a regimen based on the highest allowable dose of statin with the highest efficacy in lowering LDL-C, especially one that typically shows a reduction of LDL-C of ≡50% in normally responsive subjects. In current clinical practice, only rosuvastatin 20 mg/day or 40 mg/day and atorvastatin 40 mg/day or 80 mg/day are considered HIS therapy. In a preferred embodiment of the invention, the subject is a subject who is receiving HIS therapy and fails to achieve a 35% reduction in LDL-C, preferably a subject who receives HIS therapy and fails to achieve 30% reduction in LDL-C, 25% reduction in LDL-C, 20% reduction in LDL-C, 15% reduction in LDL-C or 10% reduction in LDL-C. In a preferred embodiment of the invention, the low responsiveness of the subject to statin therapy, in particular HIS therapy, is established after at least 1 month (continuous) HIS therapy, more preferably after at least 2 months, at least 3 months, at least 4 months, at least 5 months or at least 6 months.

Aspects of the invention defined herein relate to methods of treatment comprising administration (typically repeated administration) of a composition comprising olanzapine or a salt or solvate/hydrate of olanzapine (preferably any of the compositions previously defined herein).

Thus, in a particularly preferred embodiment of the invention, the method comprises administering the obipratropium at a dose of at least 1mg, preferably at least 2mg, at least 3mg, at least 4mg, at least 5mg, at least 6mg, at least 7mg, at least 8mg or at least 9mg, for example about 10mg, or administering a salt, solvate or co-crystal of obipratropium at an equivalent dose. According to various aspects of the invention, the method comprises administering the obipratropium at a dose of 100mg or less, more preferably 75mg or less, 50mg or less, 40mg or less, 30mg or less, 20mg or less, 15mg or less, 12.5mg or less, 12mg or less, or 11mg, or administering a salt, solvate or co-crystal of obipratropium at an equivalent dose. According to various aspects of the invention, the method comprises administering the obipratropium at a dose in the range of 1mg to 100mg, 2mg to 50mg, 3mg to 50mg, 4mg to 25mg, 4.5mg to 15mg, or 5mg to 10mg, or administering a salt, solvate, or co-crystal of obipratropium at an equivalent dose. In certain preferred embodiments, the method comprises administering the obipratropium at a dose of 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, 9mg, 10mg, 11mg, 12mg, 13mg, 14mg, 15mg, 16mg, 17mg, 18mg, 19mg, or 20mg, or administering a salt, solvate, or co-crystal of obipiproflumium at an equivalent dose. In certain particularly preferred embodiments, the method comprises administering olanzapine at a dose of 5mg, 7.5mg, 10mg, 12.5mg, or 15mg, or administering a salt, solvate, or co-crystal of olanzapine at an equivalent dose.

In a particularly preferred embodiment of the invention, the treatment comprises repeated administration of a composition comprising olanzapine or a salt, hydrate or solvate of olanzapine, preferably at a dose within the range defined previously herein. In a particularly preferred embodiment of the invention, the treatment comprises repeated administration of the composition, preferably at doses within the ranges previously defined herein, at a frequency of at least once every two days or at least once daily. In a particularly preferred embodiment of the invention, the treatment comprises repeated administration of the composition, preferably at doses as defined herein before, at a frequency of one to four times per day. In a particularly preferred embodiment of the invention, the method comprises administering a composition comprising olanzapine or a salt, hydrate or solvate of olanzapine once or twice daily, most preferably twice daily, in the dosage range defined herein above.

Thus, in a particularly preferred embodiment of the invention, the method comprises administering the olanzapine at a daily dose of at least 1mg, preferably at least 2mg, at least 3mg, at least 4mg, at least 5mg, at least 6mg, at least 7mg, at least 8mg or at least 9mg, for example about 10mg, or administering a salt, solvate or co-crystal of the olanzapine at an equivalent dose. According to various aspects of the invention, the method comprises administering the obipratropium at a daily dose of 100mg or less, more preferably 75mg or less, 50mg or less, 40mg or less, 30mg or less, 20mg or less, 15mg or less, 12.5mg or less, 12mg or less, or 11mg, or administering a salt, solvate or co-crystal of obipratropium at an equivalent dose. According to various aspects of the invention, the method comprises administering the obipratropium at a daily dose in the range of 1mg to 100mg, 2mg to 50mg, 3mg to 50mg, 4mg to 25mg, 4.5mg to 15mg, or 5mg to 10mg, or administering a salt, solvate, or co-crystal of obipratropium at an equivalent dose. In certain preferred embodiments, the method comprises administering the obipratropium at a daily dose of 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, 9mg, 10mg, 11mg, 12mg, 13mg, 14mg, 15mg, 16mg, 17mg, 18mg, 19mg, or 20mg, or administering a salt, solvate, or co-crystal of obipratropium at an equivalent dose. In certain particularly preferred embodiments, the method comprises administering olanzapine at a daily dose of 4mg, 5mg, 7.5mg, 10mg, 12.5mg, or 15mg, or a salt, solvate, or co-crystal of olanzapine at an equivalent dose.

As will be apparent to those skilled in the art, based on the teachings of the present invention, the methods of the present invention further include concurrent treatment with ezetimibe. For this purpose, ezetimibe and olanzapine (or a therapeutically acceptable salt, solvate or co-crystal of ezetimibe and olanzapine) may be administered simultaneously or about simultaneously, sequentially or simultaneously, or ezetimibe and olanzapine may be administered at different time points. In a preferred embodiment of the invention, the frequency and interval of administration of olanzapine and ezetimibe are equal, more preferably each once a day, even more preferably at the same time of day, in two separate unit dosage forms, sequentially or simultaneously, preferably in the form of a fixed dose combination product as defined herein. In preferred embodiments, the methods of the invention comprise administering ezetimibe at a daily dose of 1mg to 30mg, 2mg to 25mg, 3mg to 20mg, 4mg to 17.5mg, or 5mg to 15mg, e.g., 1mg, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, 9mg, 10mg, 11mg, 12mg, 13mg, 14mg, 15mg, 16mg, 17mg, 18mg, 19mg, or 20mg, most preferably about 10mg, or administering a salt, solvate, or co-crystal of ezetimibe at an equivalent dose.

As will be apparent to those skilled in the art, based on the teachings of the present invention, in some embodiments, the methods of the present invention further comprise concurrent treatment with HMG CoA reductase inhibitors, preferably concurrent HIS therapy. To this end, the HMG CoA reductase inhibitor and the obipratropium (or a therapeutically acceptable salt, solvate or co-crystal thereof) may be administered simultaneously or about simultaneously, sequentially or simultaneously, or may be administered at different time points. In a preferred embodiment of the invention, the frequency and interval of administration of the obipratropium and HMG CoA reductase inhibitor are equal, more preferably each administered once a day, even more preferably at the same time of day, in two separate unit dosage forms sequentially or simultaneously, or in the form of a fixed dose combination. In preferred embodiments, the methods of the invention comprise administering rosuvastatin in a daily dose of 10mg to 50mg, 15mg to 45mg, 17.5mg to 42.5mg or 20mg to 40mg, such as 15mg、16mg、17mg、18mg、19mg、20mg、21mg、22mg、23mg、24mg、25mg、35mg、36mg、37mg、38mg、39mg、40mg、41mg、42mg、43mg、44mg or 45mg, most preferably about 20mg or 40mg, or administering a salt, solvate or co-crystal of rosuvastatin in an equivalent dose. In preferred embodiments, the methods of the invention comprise administering atorvastatin in a daily dose of 30mg to 90mg, 35mg to 85mg, 37.5mg to 82.5mg, or 40mg to 80mg, such as 35mg、36mg、37mg、38mg、39mg、40mg、41mg、42mg、43mg、44mg、45mg、75mg、76mg、77mg、78mg、79mg、80mg、81mg、82mg、83mg、84mg or 85mg, most preferably about 40mg or 80mg, or administering a salt, solvate, or co-crystal of atorvastatin in an equivalent dose. In certain preferred embodiments, the methods of the invention do not include concomitant treatment with HMG CoA reductase inhibitors.

As will be apparent to those skilled in the art, the daily dosages shown herein may be included in a single unit dosage form, as well as in multiple unit dosage forms, in accordance with the teachings of the present invention. In a most preferred embodiment of the invention, the method comprises administering obipratropium (or a salt, hydrate or solvate of obipratropium) once daily at the dosages described herein. However, methods are also contemplated that include administering 2 unit dosage forms at certain predetermined times of the day, each unit dosage form including about half of a daily dose as described above, e.g., one unit dosage form administered in the morning (e.g., shortly after the subject wakes up) and another unit dosage form administered in the evening (e.g., about the time the subject spends dinner or sleeps). Embodiments are also contemplated wherein a unit dosage form comprising a higher amount of obrisedronate and/or ezetimibe than the daily dosage amounts shown herein is used. This may include, for example, the use of extended release dosage forms that remain in the body and release the active ingredient for a sustained period of time long enough.

In an embodiment, there is provided a method and/or use of a composition according to the invention, wherein the method and/or use comprises administering, preferably repeatedly administering, obipratropium and ezetimibe (or salts, hydrates or solvates of obipratropium and ezetimibe) to a subject, preferably in the form of a fixed dose pharmaceutical composition as defined herein, at a dose and frequency effective to reduce LDL-C plasma levels, apoB plasma levels and/or Lp (a) plasma levels in the subject, thereby more preferably reducing one or more of LDL-C plasma levels, apoB plasma levels and/or Lp (a) plasma levels in the subject to within the ranges described elsewhere herein. In a particularly preferred embodiment of the invention, these treatments comprise repeated administration of obipratropium and ezetimibe (or salts, hydrates or solvates of obipratropium and/or ezetimibe) according to the above-described regimen, preferably in the form of a fixed dose pharmaceutical composition as defined herein, for a period of at least one month, at least three months, at least four months, at least six months, at least nine months, at least one year, at least two years, at least three years, at least 5 years, at least 10 years, at least 20 years, at least 30 years. There is no particular upper limit and the treatment may be continued, for example, for the remainder of the subject's life, as long as the treatment is deemed beneficial to the subject's overall health and well-being (as determined by a properly qualified healthcare professional).

The pharmaceutical kit of the invention

Another aspect of the invention relates to a pharmaceutical kit comprising a package containing a plurality of unit dosage forms and a pharmaceutical instruction, wherein the unit dosage forms contain a pharmaceutical composition according to the invention, and wherein the pharmaceutical instruction contains printed instructions for directing repeated self-administration of the unit dosage forms to achieve any of the therapeutic purposes defined herein, such as the treatment and/or prevention of any of the heart diseases or dysfunctions defined herein.

According to an embodiment of the invention, the pharmaceutical kit comprises a container (e.g. a cardboard box) containing one or more blister packs (blister pack) containing a plurality of solid unit dosage forms as defined herein before, preferably a plurality of tablets as defined herein before. In particularly preferred embodiments of the invention, the pharmaceutical kit comprises at least 5, at least 8, at least 10, at least 12 or at least 15 of said unit dosage forms, for example comprising 4, 5, 6,7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 of said unit dosage forms. In one embodiment of the invention, the pharmaceutical kit comprises only unit dosage forms as defined herein containing obrisetrapib as the sole active ingredient. In one embodiment of the invention, the pharmaceutical kit generally comprises only a plurality of unit dosage forms as defined herein containing obsemipirtine (or a salt, hydrate or solvate of obsemipirtine) as the sole active ingredient, and a plurality (preferably an equal number) of unit dosage forms containing ezetimibe as the sole active ingredient, at the dosage levels described elsewhere herein. In one embodiment of the invention, the pharmaceutical kit comprises only a plurality of defined unit dosage forms, wherein each unit dosage form comprises olanzapine (or a salt, hydrate, or solvate of olanzapine) and ezetimibe (or a salt, hydrate, or solvate of ezetimibe), more preferably a plurality of fixed dose pharmaceutical compositions as defined herein. In some embodiments, the pharmaceutical kits defined herein may further comprise a plurality of unit dosage forms containing HMG CoA reductase inhibitors as the sole active ingredient, preferably atorvastatin or rosuvastatin (or salts, hydrates or solvates of atorvastatin and rosuvastatin) at the dosage levels described elsewhere herein.

According to the invention, the pharmaceutical kit comprises a pharmaceutical instruction inserted into the container, which is typically a patient information pharmaceutical instruction containing printed information, which information may include instructions on the form and composition of the unit dosage form contained in the kit, instructions on the intended therapeutic indication of the product, instructions on how the product is to be used, and information and warnings regarding side effects and contraindications associated with use. Those of ordinary skill in the art will appreciate that, based on the information provided herein, the pharmaceutical instructions as part of the kits according to the invention will generally contain information regarding therapeutic indications, uses, treatment regimens, etc. as described herein above with respect to the methods of treatment of the invention. In a particularly preferred embodiment of the invention, the pharmaceutical instructions contain printed instructions for directing repeated (self) administration of the unit dosage form for the treatment and/or prophylaxis of CVD, in particular ASCVD.

In the following, the method of treatment of the present invention based on the combination therapy of obrisetrapib and ezetimibe will be further illustrated by way of non-limiting example.

Amorphous calcium salt forms of olanzapine

In certain preferred embodiments of the present invention, the obipratropium included in the pharmaceutical composition of the present invention, in the method of the present invention, included in a unit dosage form (included in a pharmaceutical kit), etc. is in the form of a salt of obipratropium, more particularly the amorphous calcium salt of obipratropium, particularly amorphous calcium obipratropium (hemicalcium).

The amorphous obiprifflic calcium of the present disclosure is different from and distinguishable from the crystalline obiprifflic calcium disclosed in U.S. patent No. 7,872,126. A common technique for distinguishing crystalline from amorphous materials is x-ray powder diffraction. However, this technique has limitations, especially when the crystalline material is disordered. In the case of amorphous obiprift calcium, X-ray powder diffraction patterns of two different batches (lot) of amorphous obiprift calcium are provided in fig. 49 and 50. These patterns have the characteristic of the familiar "halo" type associated with amorphous materials. The X-ray powder diffraction pattern in fig. 50 has peaks at about 3.4 ° 2θ, about 7.0 ° 2θ, and about 9.2 ° 2θ. Similarly, another sample of FIG. 51 also has X-ray powder diffraction peaks at about 3.4 deg. 2 theta, about 7.0 deg. 2 theta, and about 9.2 deg. 2 theta. The X-ray powder diffraction pattern of either fig. 49 or 50 or 51 can be used to characterize amorphous obsemicalcium, however, if a sharp higher angle peak occurs occasionally, such as found at about 31.7 ° 2θ (e.g., in fig. 50), then the peak (when present) is caused by sodium chloride. In fig. 51, peaks at about 3.4 ° 2θ, about 7.0 ° 2θ, and about 9.2 ° 2θ are identified in another sample of amorphous obipratropium calcium. The peak at about 5.6 ° 2θ in fig. 51 was determined to be caused by Kapton (Kapton) foil used in the measurement setup described in example 11.20. An X-ray powder plot of crystalline obiprifflic calcium prepared in example 11.16 is shown in fig. 54. It also exhibits halo-like behaviour, which may indicate disorder for crystalline compounds.

Examples 11.18, 11.19, 11.20 and 11.21 illustrate various X-ray powder diffraction procedures. The procedure of example 11.18 was generally used to collect the data described in fig. 49, 54, 55 and 56, example 11.19 was generally used in fig. 50, example 11.20 was generally used in fig. 51, and example 11.21 was generally used in fig. 66, 67 and 68 (fig. 68 is for compound 1D instead of crystalline objected HCl).

The use of the term "amorphous" in "amorphous obiprift calcium hemi-hydrate" does not mean that the material does not have any order. As indicated by the appearance of peaks in the X-ray powder diffraction pattern, there is still some order in the sample. Thus, as used herein, the term "amorphous" in "amorphous obiprift hemi-calcium" does not mean that the X-ray powder diffraction pattern must contain only amorphous halos (but may contain halo features). In particular, this means that there is disorder, but as described below, an amorphous phase (amorphus phase) can be distinguished from a crystalline phase (CRYSTALLINE PHASE).

Another technique that may be used to distinguish crystalline from amorphous materials is polarized light microscopy (polarized light microscopy) ("PLM"). In PLM, anisotropic materials (e.g., crystalline) or isotropic materials (e.g., amorphous compounds) can be distinguished by observing the material with polarized light, and by observing the material through a cross polarizer (cross-polarizer). When exposed to polarized light passing through the crossed polarizers, the anisotropic material exhibits birefringence, as evidenced by the color change exhibited through the crossed polarizers. On the other hand, isotropic materials do not exhibit birefringence and do not exhibit a color change when exposed to polarized light.

In fig. 57, amorphous obipratropium calcium was analyzed by polarized light microscopy as described in example 11.17. As shown in fig. 57, the material under study did not undergo birefringence, indicating that the material was amorphous. In contrast, fig. 58 is a polarized light micrograph of crystalline obipratropium calcium prepared according to example 11.16. Notably, the particles shown in fig. 58 (which are black and white) exhibit brighter contrast. In a corresponding color version, the graphic is multi-colored. Thus, fig. 58 shows that there is crystallinity. In addition, the crystals in fig. 58 are larger than the particles provided in the amorphous obiprifleside calcium polarized light micrograph of fig. 57. Thus, PLM and/or lack of birefringence can be used to characterize amorphous obiprift calcium hemi-hydrate.

Other techniques may be further used to distinguish amorphous obiprift calcium from crystalline obiprift calcium and thus other techniques may be used to characterize amorphous obiprift calcium. One such technique is modulated DIFFERENTIAL SCANNING differential scanning calorimetry (mDSC), also known as "mDSC". The difference in heat required to raise the temperature of the sample compared to the reference is measured as a function of temperature and can be measured using modulated differential scanning calorimetry (mDSC). In the mDSC thermogram (thermogram), the glass transition temperature, which can be used to characterize amorphous materials, can also be measured. In fig. 60 (the procedure of which is set forth in example 11.25), the mDSC thermogram of amorphous obiprift hemicalcium was measured using an open sample holder, enabling volatile gases to escape during the measurement. In fig. 60, the opening is formed by piercing a lid on a disk vessel (pan) to create a pinhole. The glass transition temperature of this sample was recorded to be about 110 ℃.

With respect to thermal measurements, the term "about" generally refers to a variability of ±1 ℃. In contrast, crystalline obipratropium calcium has a higher glass transition temperature under the same conditions, and three measurements in fig. 62 indicate a range between about 118 ℃ and about 125.5 ℃. In some embodiments, the amorphous obiprift calcium has a glass transition temperature between about 109 ℃ and 112 ℃ when measured with pinholes. In one sample, at example 11.26, amorphous obiprift calcium has been found to have a glass transition temperature of about 111 ℃ (111.32 ℃ at the midpoint) and is shown in fig. 61. The initial temperature was measured to be about 102 ℃ (101.62 ℃) and the end temperature was measured to be about 118 ℃ (117.58 ℃).

The glass transition temperature of amorphous obiprift calcium can also be measured using a closed disk vessel using an mDSC. The type of sample preparation may affect the measured glass transition temperature. In such a case, the glass transition temperature decreases to a temperature below about 100 ℃ and in particular between about 70 ℃ and about 92 ℃ depending on the humidity.

Other thermal techniques may also be used to analyze and characterize amorphous obiprift calcium, for example using thermogravimetric analysis (thermogravimetric analysis, TGA). Fig. 59 is a thermogram of amorphous obiprift calcium showing less than 1% weight loss when heated to about 200 ℃. Such weight loss may be between, for example, about 0.8% and about 0.95% (including between about 0.84% and about 0.92%). In fig. 59, the weight loss was determined to be about 0.85%. The water content of this particular material was found to be about 1.5%. In some embodiments, the water content may be higher and include a range of about 0 wt% to about 5 wt%, including up to about 4 wt%, up to about 3 wt%, and between about 0.5 wt% and 1.5 wt%.

Solid-state ¹³ C-NMR spectroscopy is another technique that can be used to characterize amorphous materials. Fig. 63 shows solid-state ¹³ C-NMR spectra of both crystalline and amorphous obiprift calcium, wherein fig. 64 and 65 show crystalline and amorphous obiprift calcium, respectively. There are at least two differences in the spectrum. The crystalline phase has a peak at about 22.1ppm, while the peak is absent in the amorphous phase. Furthermore, peaks at about 29.5ppm are evident in the crystalline phase, while the peaks are not so evident in the amorphous phase. Thus, the absence of a solid-state ¹³ C-NMR peak at about 22.1ppm and/or a distinct peak at about 29.5ppm can be used to characterize amorphous obsemicalcium. In addition, amorphous obiprift calcium may be characterized using a solid state ¹³ C-NMR spectrum substantially the same as that shown in fig. 65. The absence of peaks in this context does not mean that there must be no intensity, for example 22.1ppm or 29.5ppm, but that the intensity is not as pronounced as in the C-NMR spectrum of crystalline obiprift semi-calcium ¹³.

The properties of crystalline materials also typically differ from those of amorphous materials. Thermodynamically, crystalline materials are more physically stable than amorphous materials. Thus, there is a thermodynamic driving force for converting amorphous compounds into crystalline compounds. Under accelerated stress conditions, if there is a physical transformation of the solid form, it is therefore generally expected that the physical transformation will be from amorphous to crystalline. However, the situation is the opposite for obiprift calcium.

Fig. 54 is a diagram of an X-ray powder diffraction pattern of the crystalline calcium obiprift trapib obtained, and fig. 55 is a diagram of an X-ray powder diffraction pattern of the crystalline calcium obiprift trapib obtained under stress conditions. In fig. 55, four diffraction patterns are shown based on the stability study described in example 11.27. Profile 1 is an X-ray powder diffraction pattern of a sample of amorphous obiprifflic calcium. Profile 2 is an X-ray powder diffraction pattern of a sample of crystalline obiprifflic calcium. In profile 3, a sample of crystalline obipratropium calcium is exposed to 70 ℃ for one day at 75% relative humidity. As can be seen from pattern 3, the X-ray powder diffraction pattern shows almost complete loss of crystallinity within the day. After 7 days under the same conditions, the results were still the same as those seen in pattern 4. A similar experiment was performed on amorphous obiprifflic calcium as shown in fig. 56. Profile 1 is the stability obtained prior to sample placement. Exposure of the material to the same 70 ℃ and 75% relative humidity conditions did not trigger crystallization and the material remained amorphous after 7 days (profile 2) and after 14 days (profile 3). Thus, these experiments show that, contrary to one's expectation, the amorphous form of the calcium obipratropium is more stable than the crystalline calcium obipratropium.

In some embodiments of the present disclosure, stable amorphous obiprift calcium is provided herein. In these examples, amorphous calcium obipratropium has a greater physical stability than crystalline calcium obipratropium under typical pharmaceutical use and processing conditions.

While not wishing to be bound by theory, the kinetics herein are such that the amorphous phase is kinetically stable relative to the thermodynamically more stable crystalline phase, at least under drug related processing and use conditions. The result of this stability profile is that amorphous obiprift calcium is more suitable for drug development and use than the corresponding crystalline phase. Although physically more elastic, amorphous obiprifflic calcium is more soluble than highly insoluble crystalline obiprifflic calcium.

The solubility of olanzapine is particularly challenging. For example, at 20 ℃, the solubility of obiprifepin in water has been measured to be substantially less than 0.1mg/mL. It is desirable to have a solid form of obipratropium that will deliver a greater amount of obipratropium.

Although solubility is a thermodynamic quantity of a material (thermodynamic quantity), the dynamic solubility of a material can be measured without necessarily achieving thermodynamic equilibrium. Such measurements provide solubility under metastable conditions and provide information, for example, about the amount of material that dissolves over time.

The amorphous form has a higher dynamic solubility and dissolution rate than the crystalline form (as well as by extension of the obipratropium itself). As described in example 11.28, dynamic solubility measurements of both crystalline and amorphous calcium obiprifleside were performed in a biologically relevant medium at different pH (i.e., at a pH of about 5.0 (FeSSIF conditions) and at a pH of about 6.5 (FaSSIF conditions).

Table W shows the measured solubility of two different batches of amorphous and crystalline obiprift-calcium in FeSSIF medium at 37 ℃ over the course of 2 hours. In both cases, at all measured time points, amorphous obipratropium calcium has a higher concentration in solution than the corresponding crystalline material. The concentrations in table W are those of obrisetrapib (i.e., free acid).

Table W-dynamic solubility of crystalline and amorphous calcium obiprift trapezole in FeSSIF (pH 5.0) at 37 °c

Table X shows a similar experiment at 37 ℃, but in a FaSSIF medium at pH 6.5. As with table W, amorphous obipratropium calcium has a higher concentration in solution than the corresponding crystalline material in both batches at all measured time points. The concentrations in table X are those of obrisetrapib (i.e., free acid).

Table X-crystalline calcium obipriftrapib and amorphous ozagrel at 37C dynamic solubility of Bicetrapib-hemi-calcium in FaSSIF (pH 6.5)

Since amorphous obipratropium calcium dissolves faster than the corresponding crystalline phase, more drug is available for immediate use and bioavailability in the amorphous phase may be higher than in the crystalline phase.

Unlike many amorphous organic compounds, amorphous obiprift calcium is not easily absorbed with moisture and is therefore also advantageous. For example, when exposed to a relative humidity approaching 90%, the measured moisture absorption is typically less than about 5%. This lack of hygroscopicity is advantageous because it does not require any special handling or storage conditions. Similarly, there are no other drawbacks typically associated with the manufacture and use of amorphous materials. For example, amorphous materials often present challenges in achieving chemical purity. Here, however, amorphous obipratropium calcium can be prepared in a conventional manner with a chemical purity of 99.9% or more.

In some embodiments of the present disclosure, substantially pure amorphous obipratropium calcium is provided. In these and other embodiments, the substantially pure amorphous obiprift calcium has a chemical purity of 99.9% or greater.

In many aspects of the present disclosure, a process for preparing amorphous calcium salts of olanzapine, such as amorphous olanzapine calcium salt, is provided, wherein the process comprises treating the olanzapine with an acid to form a salt, solvate, or composition, separating the resulting salt, solvate, or composition, and treating the salt, solvate, or composition with a calcium source to form amorphous calcium salt of olanzapine, such as amorphous calcium olanzapine. The resulting salt may then be isolated.

Examples of the calcium source include calcium salts such as calcium halide salts and soluble calcium salts. In many embodiments, the calcium source is calcium chloride.

It has been found that when an intermediate salt, solvate or composition is present (such composition comprising the corresponding acid used to prepare the salt), an amorphous salt of obipratropium calcium, such as amorphous obipratropium calcium half-hydrate, is prepared. Direct treatment of olanzapine with a calcium base (e.g., calcium hydroxide) has not been found to be a viable method for preparing amorphous salts of olanzapine calcium due to low solubility, weak points of available base, or both. In contrast, it has been found that by configuring an intermediate salt (e.g., sodium salt), it is possible to prepare amorphous obiprifleside calcium. However, even in the case of the sodium salt, it is preferable to utilize additional salts or salt form exchanges (e.g., use of a composition or solvate rather than the actual salt) associated with the sodium salt of obsemitrapib for purity and yield purposes. In particular, the use of a salt, solvate or composition enables the production of amorphous calcium salts of obipratropium, such as amorphous obipratropium calcium half-hydrate, in high purity.

Exemplary salts that may be prepared as intermediates include salts from sulfonates (e.g., benzenesulfonate, toluenesulfonate, naphthalenesulfonate, camphorsulfonate, ethanesulfonate, ethanedisulfonate (edisylate) or methanesulfonate), sulfates (e.g., methylsulfate), halogens (e.g., chloride, iodide or bromide), acetates, aspartate, benzoate, bicarbonate, acid tartrate (bitartrate), carbonates, citrates, caprates, fumarates, glucoheptonate, gluconate, glutamate, glycolate, caproate, hydroxynaphthoates (hydroxynaphthoate), isethionates, lactates, malates, maleates, mandelates (mandelate), muciates (mucate), nitrates, octanoates, oleates, pamoate, pantothenates, phosphates, polygalacturonates, propionates, salicylates, stearates, succinates, tartrates or theates (teoclates). When an intermediate system solvate or composition, then the corresponding acid may be used or present. In addition, when the intermediate system is a solvate, the intermediate may further include a solvent (e.g., an organic solvent) or water, in which case the solvate will be a hydrate. One such organic solvent is cyclopentyl methyl ether (cyclopentyl METHYL ETHER, CPME).

In some embodiments, the intermediate system acid is a solvate. In these and other embodiments, a solvate of an intermediate systematic acid and an organic solvent. In some particular embodiments, the intermediate system comprises a solvate of an acid and a solvent. In some of these embodiments, the acid is hydrochloric acid and the solvent is CPME.

In many aspects of the disclosure, the disclosure includes methods for preparing amorphous calcium salts of obipratropium, such as amorphous calcium obipratropium. The disclosure further includes amorphous calcium salts of obiprifepin, including amorphous calcium obiprifepin. In one such preparation, an intermediate referred to herein as crystalline obiprift HCl is used in the preparation of amorphous obiprift calcium (e.g., amorphous obiprift calcium hemihydrate).

In many aspects of the present disclosure, amorphous obipratropium calcium is prepared by chemical synthesis using an intermediate represented by formula (IH):

Wherein y varies such that the mass percent of HCl varies from 0.01 wt% to 8 wt% and is believed to more include the relevant organic solvent, for example, in the form of a solvate. In some embodiments, y varies from 0.002 to 1.5. In some embodiments, y varies from 0.3 to 1. In some embodiments, y varies from 0.4 to 0.6 (including between 0.5 and 0.6). In some embodiments, formula (IH) as a solvate is isolated in its crystalline form. In many embodiments, the solvent is CPME. Other solvents that may form solvates include toluene and heptane.

The olanzapine HCl typically prepared herein is crystalline. Furthermore, when CPME is used to prepare crystalline obiprift HCl, the term crystalline obiprift HCl may include CPME as a solvate. In formula (IH), the solvate is a solvate of an organic solvent, and in many embodiments, the solvent is CPME. In some embodiments, the present disclosure provides compositions comprising crystalline obiprift HCl.

Formula (IH) is referred to as obiprift HCl and when it is crystalline, it is referred to as crystalline obiprift HCl.

Without being bound by theory, it is believed that crystalline HCl obipratropium is a mixed salt solvate. It has been found that when CPME is used to deliver HCl in the reaction that produces formula (IH), the chloride content of formula (IH) ranges between about 2.5 wt.% and 3.0 wt.%, which is lower than one would expect for neutral salts, i.e., about 4.8 wt.%.

In many embodiments, when CPME is so used, it is found in the material as it crystallizes. When CPME is used in the reaction to deliver dry HCl and is thus found in crystalline materials, the resulting crystalline formula (IH) material is referred to as crystalline obrisetrapezil HCl, and these X-ray powder diffraction patterns are visible in fig. 66. The advantage of using crystalline obiprift HCl as an intermediate is that the chemical purity of the resulting amorphous obiprift calcium is typically 99.9% or higher. Chemical purity is a quantitative representation of the presence or absence of other chemical entities in addition to the compound being tested. For example, amorphous obiprift calcium of chemical purity 99.9% means that no more than 0.1% of the compounds in the sample of amorphous obiprift calcium are other entities. Physical purity refers to the amount of other solid forms of the same compound present, in the case of amorphous obiprifflic calcium, the other solid form is crystalline obiprifflic calcium. The disclosure herein provides amorphous obiprift calcium that is physically pure, meaning that the amorphous obiprift calcium is free or substantially free of crystalline obiprift calcium. Unless otherwise indicated herein, purity measurements provided herein are measurements of chemical purity.

The HCl obiprifepin as used herein is not limited to crystalline obiprifepin HCl. In fact, crystalline obiprift HCl may become amorphous upon desolvation.

Under stress, crystalline obiprift HCl loses its crystallinity. In fig. 66, profile 2 reflects crystalline obiprift HCl subjected to a gentle drying treatment thereby removing surface solvent, and it can be seen that this compound is crystalline. In contrast, the samples whose x-ray powder diffraction was measured in profile 1 were subjected to a stronger drying treatment at 55 ℃ and a pressure of 2mbar for 48 hours. It is apparent that such drying may cause the material to self-crystallize to become amorphous due to HCl loss and CPME desolvation. For example, using ¹ H-NMR spectroscopy showed the presence of CPME in the top profile, but virtually no CPME in the lower amorphous profile. Thus, the amorphous pattern represents HCl obipratropium that is not crystalline obipratropium. It may be obipratropium but is believed to have HCl bound to obipratropium as a solvate and thus be HCl obipratropium but has a lower chloride content than is typically found in the range of crystalline obipratropium HCl. In some embodiments, the chloride content is less than 0.1 wt%, for example between about 0.01 wt% and 0.1 wt%.

Crystalline olanzapine HCl may be characterized by an X-ray powder diffraction pattern comprising peaks at about 9.8 deg. 2 theta. In some embodiments, crystalline obiprap HCl may be characterized by an X-ray powder diffraction pattern comprising one or more peaks at about 8.1 ° 2θ, about 9.8 ° 2θ, about 13.8 ° 2θ, about 16.7 ° 2θ, or about 19.5 ° 2θ. Table Y provides exemplary peaks that may be present in crystalline obiprift HCl. In some embodiments, crystalline obiprap HCl may be characterized by an X-ray powder diffraction pattern substantially the same as in fig. 67, but it is believed that the material analyzed in fig. 67 is measured in such a way that no peak between about 4.3 ° 2θ and about 4.7 ° 2θ is measured.

Table Y

°2θ	Strength of
		8.1	950
9.8	1650
		13.8	2000
16.7	2900
		19.5	4100
21.1	3700
		21.6	3800
22.4	3600
		24.9	1950
26.6	1850

Another intermediate of formula (VI) of the intermediate system for the preparation of olanzapine

Wherein Y ¹ is a protecting group (e.g., as described herein), A ^n- is an anion, and n is an integer from 1 to 3.

In one embodiment, the compound of formula (VI) is a mesylate salt, wherein n is1, y ¹ is t-butyl, and has the structure of compound 1D:

The ¹ H-NMR spectrum of compound 1D (in solution) can be found in FIG. 69. Crystalline compound 1D may be characterized by an X-ray powder diffraction pattern comprising one or more peaks at about 5.2 ° 2θ or about 9.1 ° 2θ. In some embodiments, crystalline compound 1D may be characterized by an X-ray powder diffraction pattern comprising one or more peaks at about 5.2 ° 2θ, about 9.1 ° 2θ, about 15.9 ° 2θ, about 16.5 ° 2θ, about 17.2 ° 2θ, about 18.6 ° 2θ, and about 19.2 ° 2θ. Table Z provides exemplary peaks that may be present in crystalline compound 1D (no peak at about 5.2 ° 2θ was measured due to instrument limitations in the reflective mode). In some embodiments, crystalline compound 1D may be characterized by an X-ray powder diffraction pattern substantially the same as figure 68.

Table Z

Crystalline compounds (e.g., crystalline compound 1D and crystalline obiprift HCl) may be characterized, for example, by x-ray powder diffraction. The X-ray powder diffraction pattern is an X-y plot with °2θ (diffraction angle) on the X-axis and intensity on the y-axis. The peak is typically represented and referenced by its position on the x-axis rather than its peak intensity on the y-axis, as the peak intensity may be particularly sensitive to sample orientation (see Pharmaceutical Analysis [ drug analysis ], lee and Web, pages 255 to 257 (2003)). Thus, intensity is not generally used to characterize solid morphology. Data from X-ray powder diffraction can be used in a variety of ways to characterize crystalline forms. For example, the full X-ray powder diffraction pattern output from the diffractometer can be used to characterize crystalline olanzapine HCl compound or crystalline compound 1D. However, smaller subsets of such data may also be suitable, and generally suitable, for characterizing such compounds. For example, a collection of one or more peaks from such a profile may be used to so characterize these compounds. When the phrase "one or more peaks" in a column of peaks is provided from an X-ray powder diffraction pattern, it is generally meant that any combination of the listed peaks can be used for characterization. Furthermore, the fact that there are other peaks in the X-ray powder diffraction pattern typically does not negate or otherwise limit the characterization.

In addition to the variability in peak intensity, there may be variability in peak position on the x-axis. However, such variability is often considered when reporting the location of peaks for characterization purposes. Such variability in peak position along the x-axis may originate from several sources (e.g., sample preparation, particle size, moisture content, solvent content, instrument parameters, data analysis software, and sample orientation). For example, samples of the same crystalline material prepared under different conditions may produce slightly different diffraction patterns, and different X-ray instruments may operate with different parameters, and such factors may result in slightly different diffraction patterns from the same crystalline solid. Because of the source of such variability, the word "about" is often used to describe the X-ray diffraction peak before the peak in °2θ. For purposes of reporting data herein, the values are typically ±0.2° 2θ, and whenever disclosed herein, the term "about" is intended to report the values with such variability, whether or not present. In some cases, variability may be higher, depending on instrument conditions, including maintenance of the instrument.

In some embodiments, crystalline compound 1D may be further characterized by an X-ray powder diffraction pattern substantially the same as the X-ray powder pattern of fig. 68.

In many aspects of the present disclosure, there is provided a process for preparing an amorphous calcium salt of obipratropium, such as amorphous obipratropium hemi-calcium, wherein the process comprises:

i. treating the obipratropium with HCl to obtain crystalline obipratropium HCl;

separating the crystalline obiprift HCl;

Preparing an amorphous calcium salt of olanzapine, such as amorphous olanzapine calcium salt, from crystalline olanzapine HCl separated in step (ii), and

Separating the amorphous calcium salt of obiprifepin, e.g. amorphous obiprifepin-hemi-calcium.

In other aspects of the present disclosure, there is provided a method of preparing obipratropium, wherein the method comprises:

(a) Preparing a compound of formula (IV) by coupling a compound of formula (II) or a salt thereof with a compound of formula (III);

wherein X ¹ is a leaving group and Y ¹ is a protecting group;

(b) The carbamate of formula (V) is prepared from the compound of formula (IV) and isolated as a solid salt of formula (VI):

Wherein Y ¹ is a protecting group, a ^n- is an anion, and wherein n is an integer from 1 to 3;

(c) Optionally desalting the compound of formula (VI) and alkylating with the compound of formula (VII) to provide the compound of formula (VIII):

Wherein X ² is a leaving group, Y ¹ is a protecting group, and

(D) Converting the compound of formula (VIII) to obiprap, wherein the reaction steps (a) to (d) are performed in an organic solvent, optionally without isolation of compounds (IV), (V) and (VIII) in an organic solvent, and wherein the process does not need to include chromatography.

The reactions in steps (a) to (d) of the subject process are carried out in solvents and if the intermediate compounds of formulae (IV), (V) and (VIII) are to be further processed into the final product, it is not necessary to separate the intermediate compounds of formulae (IV), (V) and (VIII) from their respective solvents. This means that any solvent exchange between reaction steps (x) and (x+1) is performed by evaporating at least a part of the solvent used in step (x) and gradually adding the solvent of step (x+1) so that the compound remains in solution during the solvent exchange. The intermediate compound of formula (VI) may be isolated from the solvent as a salt in solid form, such that the intermediate compound of formula (VI) may be washed to remove impurities. This separation step ensures adequate purity of the downstream product. The subject process need not include a purification step using chromatography (e.g., column chromatography) to achieve the chemical purity levels described herein.

Method for preparing amorphous calcium salts such as amorphous obiprift calcium salt-steps (i) to (ii) of aspects (i) to (iv)

In some embodiments of the process for preparing amorphous calcium salts of olanzapine (e.g., amorphous olanzapine calcium salt) the process comprises the step (i) of treating olanzapine with HCl in an organic solvent to obtain crystalline olanzapine HCl.

In some embodiments, the crystalline obiprift HCl has a purity of 98% or greater, e.g., 98.5% or greater, 99% or greater, 99.5% or greater, or even greater.

In some embodiments, the HCl in step (i) is in a suitable solvent. Such solvents may be aqueous or organic. In some embodiments, the organic solvent used in step (i) comprises a mixture of solvent and anti-solvent. In some embodiments, the solvent is selected from the group consisting of methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl tert-butyl ether, cyclopentyl methyl ether, N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1, 4-dioxane, 1, 2-difluorobenzene, toluene, hexafluoroisopropanol, and water. In some embodiments, the antisolvent is selected from the group consisting of n-heptane, n-hexane, n-pentane, and cyclohexane.

In some embodiments, HCl has sufficient solubility in the anti-solvent such that it can be used as a suitable solvent. In some embodiments, the organic solvent used in step (i) comprises a mixture of cyclopentyl methyl ether and n-heptane. In some embodiments, the organic solvent used in step (i) further comprises toluene. In some embodiments, toluene is the major component of the organic solvent.

In some embodiments, step (i) comprises providing obetatrapib in a mixture of cyclopentyl methyl ether and n-heptane, increasing the temperature to between 35 ℃ and 40 ℃ while stirring, adding dry HCl to the cyclopentyl methyl ether and increasing the temperature again to between 50 ℃ and 55 ℃, and then adding additional n-heptane as an anti-solvent. At this point, a small portion of the reaction mixture may optionally be extracted and cooled to a temperature between 10 ℃ and 15 ℃ to obtain a slurry of crystals of crystalline obrisetrapezil HCl in a mixture of cyclopentyl methyl ether and n-heptane (referred to herein as a "seed slurry"). Optionally, all or a portion of the seed slurry of crystalline olanzapine HCl may then be added back to the reaction mixture. The seed aids in nucleation, but is not required. The resulting reaction mixture is then cooled to a temperature between 5 ℃ and 15 ℃ (e.g., 10 ℃ to 15 ℃) and then the crystalline obiprift HCl is crystallized from the system while stirring. In some embodiments, the crystalline obiprap HCl is crystallized over a period of 12 hours or more, then filtered (e.g., by a filter dryer), subjected to one or more optional washing steps (e.g., washing with a mixture of cyclopentyl methyl ether and n-heptane), and dried. In some cases, the wet cake of crystalline olanzapine HCl is vacuum dried in multiple steps using temperatures of 25 ℃ to 30 ℃, 30 ℃ to 40 ℃, 40 ℃ to 50 ℃, then 50 ℃ to 55 ℃, e.g., 25 ℃, 35 ℃, 46 ℃ and 54 ℃.

In some embodiments, the method of preparing crystalline obiprift HCl includes adding seed crystals (e.g., as a seed slurry). Seed crystals of HCl compound can be formed into a slurry by following step (i) set forth above, and after addition of dry HCl to cyclopentyl methyl ether and anti-solvent n-heptane, extracting a small portion of the reaction mixture, cooling it to a temperature between 10 ℃ and 15 ℃, thereby providing a slurry of crystals of crystalline obstopper HCl in cyclopentyl methyl ether and n-heptane.

Thus, in one embodiment, step (i) comprises providing crystalline obetatrapezil HCl in a mixture of cyclopentyl methyl ether and n-heptane, increasing the temperature to between 35 ℃ and 45 ℃ while stirring, adding dry HCl to the cyclopentyl methyl ether and increasing the temperature again to between 50 ℃ and 55 ℃, adding additional n-heptane as an antisolvent, and optionally adding seeds of HCl compound (e.g., as a seed slurry prepared as described herein), cooling to a temperature between 5 ℃ and 15 ℃ (e.g., 10 ℃ to 15 ℃) and subsequently crystallizing crystalline obetatrapezil HCl from the system while stirring. In some embodiments, the crystalline obiprift HCl is crystallized over a period of 12 hours or more, then filtered, subjected to one or more optional washing steps (e.g., washing with a mixture of cyclopentyl methyl ether and n-heptane), and dried. In some embodiments, the crystalline obiprapib HCl is dried in vacuo. In some embodiments, the crystalline obiprift HCl is dried in a vacuum oven at 25mbar pressure and a temperature of 55 ℃ for 10 hours or more. In some embodiments, after the drying procedure, the crystalline obiprift HCl includes less than 0.1 weight percent residual cyclopentyl methyl ether.

In some embodiments, step (i) comprises providing a solution of obiprift in cyclopentyl methyl ether at a concentration of between 30 and 40 weight percent, e.g., 33 to 37 weight percent, less than 1 weight percent of the first organic solvent (e.g., toluene) used in step (d), less than 1 weight percent of n-heptane based on the weight of the solution, adding n-heptane, increasing the temperature to 35 to 45 ℃ while stirring, adding dry HCl in cyclopentyl methyl ether and increasing the temperature again to 50 to 55 ℃, adding additional n-heptane as an antisolvent, optionally adding seeds of crystalline obiprift HCl (e.g., as seed slurry as prepared herein), cooling to a temperature of between 10 and 15 ℃, then crystallizing the crystalline obiprift HCl from the system while stirring, e.g., during a period of at least 12 hours, followed by filtration, drying with the cyclopentyl methyl ether, and drying the mixture or mixtures thereof, e.g., in vacuo. In some embodiments, the amount of toluene is significantly greater.

In some embodiments, the crystalline obiprift HCl from step (i) is separated in step (ii). In some embodiments, the isolated crystalline obiprift HCl has a purity of 98% or greater, e.g., 98.5% or greater, 99% or greater, 99.5% or greater, 99.7% or even greater.

Another embodiment of the present disclosure relates to crystalline obiprift HCl obtained or obtainable by the process defined herein.

Yet another embodiment of the present disclosure relates to HCl obiprifepin, including crystalline obiprifepin HCl.

In some embodiments, crystalline obiprap HCl is stored at controlled room temperature and under nitrogen atmosphere and protected from moisture, thereby preventing the formation of amorphous solids, e.g., by desolvation.

Method for preparing amorphous calcium salts of obiprifepin, such as amorphous obiprifepin semi-calcium-steps (iii) to (iv) of aspects (i) to (iv)

In some embodiments of the process for preparing amorphous calcium salts of olanzapine (e.g., amorphous olanzapine calcium) the process comprises steps (iii) through (iv) preparing amorphous calcium salts of olanzapine from the crystalline olanzapine HCl separated in step (ii) and isolating the amorphous calcium salts of olanzapine (e.g., amorphous olanzapine calcium).

In some embodiments of the method of separating amorphous calcium salt of obipsilate according to step (iv), the amorphous calcium salt of obipsilate is in the form of amorphous obipsilate hemi-calcium:

in some embodiments of the method of preparing obipratropium, step (iii) comprises the steps of:

(iii-1) converting the crystalline obiprifepin HCl of step (ii) to provide obiprifepin in an organic solvent;

(iii-2) treating the olanzapine in the organic solvent with aqueous sodium hydroxide solution to form the sodium salt of the olanzapine, and

(Iii-3) treating the sodium salt of obipratropium with an aqueous solution of calcium chloride to form amorphous obipratropium calcium chloride;

Wherein the compounds in steps (iii-1) and (iii-2) are not isolated.

Thus, in some embodiments, step (iii-1) comprises the steps of:

(aa) providing the crystalline obiprifepin HCl isolated in step (ii);

(bb) dissolving crystalline olanzapine HCl in a mixture of water and isopropyl acetate while stirring. In some embodiments, step (bb) is performed at a temperature between 15 ℃ and 25 ℃;

(cc) allowing phase separation and subjecting the resulting organic phase to one or more subsequent washing steps with water, wherein after each washing step an aqueous phase is separated, thereby obtaining a washed organic phase, and

(Dd) performing two or more distillations on the washed organic phase resulting from step (cc) at a temperature of 50 ℃ or less (e.g., 30 ℃ or less), with the intermediate addition of ethanol, thereby obtaining a solution of obpezotezole in ethanol.

In some embodiments, step (iii-2) comprises the steps of:

(ee) adding an aqueous NaOH solution to the solution obtained in step (dd) and stirring the resulting mixture at a temperature of, for example, between 20 and 25 ℃ for at least 4 hours to obtain a solution of the sodium salt of obipratropium, and

(Ff) optionally filtering the solution obtained in step (ee).

In some embodiments, step (iii-3) comprises the steps of:

(gg) preparing a CaCl ₂ solution by adding deionized water to CaCl ₂ while stirring, followed by adding ethyl acetate as a co-solvent, and stirring the resulting mixture for 10 minutes to 30 minutes;

(hh) cooling the CaCl ₂ solution obtained in step (gg) to a temperature of 8 ℃ to 12 ℃ and adding to the solution obtained in step (ff) or (ee) via a filter while stirring at said temperature;

(ii) Stirring the slurry from step (hh) for about 1 hour to about 10 hours. In some embodiments of step (ii), the stirring is performed at a temperature between 8 ℃ and 12 ℃;

(jj) separating solids from the slurry obtained in step (ii) by filtration. In some embodiments of step (jj), the separating is performed at a temperature between 8 ℃ and 12 ℃;

(kk) washing the filter residue obtained in step (jj) with water in one or more washing steps. In some embodiments of step (kk), the washing is performed at a temperature between 8 ℃ and 12 ℃, and

(Ll) the washed residue obtained in step (kk) is dried in vacuo, for example at a temperature of 40 ℃ to 50 ℃ for more than 16 hours (e.g. 50 hours, 100 hours, 150 hours or 200 hours, or even more) to obtain amorphous obiprift calcium (sometimes also referred to herein as compound 3).

In some embodiments, the amorphous obiprift calcium is subjected to a subsequent reprocessing procedure. In some embodiments, amorphous obsemicalcium is further reprocessed by dissolving in ethanol (e.g., twice the weight of the ethanol relative to amorphous obsemicalcium) at a temperature of 25 ℃ to 50 ℃, followed by cooling to 10 ℃ to 15 ℃, followed by filtration into a mixture of aqueous calcium chloride solution and ethyl acetate that is also cooled to 10 ℃ to 15 ℃, followed by filtration, washing with water and vacuum drying at a temperature of 45 ℃ or less for 20 hours or more.

In some embodiments of step (iv), amorphous calcium ethyltrapezium having a purity of 95% or higher, for example a purity of 95.5% or higher, 96% or higher, 96.5% or higher, 97% or higher, 97.5% or higher, 98% or higher, 98.5% or higher, 99% or higher, 99.1% or higher, 99.2% or higher, 99.3% or higher, 99.4% or higher, 99.5% or higher, 99.6% or higher, 99.7% or higher, 99.8% or higher, or 99.9% or higher is isolated.

In some embodiments, amorphous obiprift calcium is ground. In some embodiments, the milling process is adapted (e.g., parameters such as feed rate, venturi pressure, and milling pressure are adapted) to enable production of micronized amorphous obipratropium calcium.

Method for preparing obipratropium-step (a) of aspects (a) to (d)

In step (a) of the process for preparing obipratropium according to the present disclosure, a compound of formula (II) or a salt thereof is coupled with a compound of formula (III) to provide a compound of formula (IV) (wherein, for example, X ¹ is a leaving group and Y ¹ is a protecting group as described herein).

Step (a) of the subject process starts from compound (2 r,4 s) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline) of formula (II) or a salt thereof:

the compounds of formula (II) may be obtained, for example, using the procedure disclosed in WO 2016/024858 A1 or WO 2007/116922 A1, both of which are incorporated herein by reference in their entirety. In some embodiments, the compound of formula (II) may be obtained from the stable corresponding salt, and may be obtained in pure and solid form. The solid form may be amorphous or crystalline. In some embodiments, the compound of formula (II) is obtained from the corresponding crystalline salt.

In some embodiments, the compound of formula (II) provided in step (a) is a salt of formula (IIA) or (IIB):

wherein a ^m- is an anion and n is an integer from 1 to 3.

In some embodiments, the compound of formula (II) provided in step (a) is a salt of formula (IIA). In some embodiments, the compound of formula (IIA) is used directly in the coupling reaction with the compound of formula (III) without performing a desalting step.

In some embodiments, the compound of formula (II) provided in step (a) is a salt of formula (IIB). In some embodiments, the compound of formula (IIB) is used directly in the coupling reaction with the compound of formula (III) without performing a desalting step.

In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIA) or (IIB). In some embodiments, the coupling reaction of step (a) is preceded by the steps of:

(pre-a 1) providing a compound of formula (IIA) or (IIB):

(pre a 2) desalting the compound of formula (IIA) or (IIB) to obtain the compound of formula (II),

Wherein the reaction in step (pre-a 2) is performed in an organic solvent, the compound of formula (II) is not isolated in the organic solvent, and the process does not comprise chromatography.

In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIA). In some embodiments, the compound of formula (II) in step (a) is obtained from a salt of formula (IIB).

In some embodiments, the salt of formula (IIA) or (IIB) is selected from salts having an anion A ^m- selected from sulfonate (e.g., benzenesulfonate, toluenesulfonate, naphthalenesulfonate, camphorsulfonate, ethanesulfonate, ethanedisulfonate, or methanesulfonate), sulfate (e.g., methylsulfate), halogen (e.g., chloride, iodide, or bromide), acetate, aspartate, benzoate, bicarbonate, acid tartrate, carbonate, citrate, decanoate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, hexanoate, hydroxynaphthoate, hydroxyethylsulfonate, lactate, malate, maleate, mandelate, muciate, nitrate, octoate, oleate, pamoate, pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, succinate, tartrate, and theachloride.

In some embodiments, the salt of formula (IIA) or (IIB) is selected from salts having an anion a ^m- selected from chloride, bromide, acid tartrate, sulfate, and sulfonate.

In some embodiments, the salt of formula (IIA) or (IIB) is selected from salts having an anion a ^m- selected from chloride, bromide, acid tartrate and methanesulfonate.

In some embodiments of the salts of formula (IIA) or (IIB), m is 1.

In some embodiments, the salt has formula (IIA) and anion a ^m- is methanesulfonate, where m is 1. Methanesulfonic acid (MSA) salt (also referred to herein as compound 1A, shown below) is obtainable by the process disclosed in WO 2016/024858A1 or WO 2007/116922 A1, both of which are incorporated herein by reference in their entirety.

In some embodiments, the desalting of the compound of formula (IIA) or (IIB) in step (pre-a 2) is performed in a mixture of aqueous sodium hydroxide solution and an organic solvent selected from toluene, methylene chloride, cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, chlorobenzene, and combinations thereof, followed by heating the mixture, then cooling the mixture, and allowing the system to phase separate and separate the aqueous phase. In some embodiments, the solvent is toluene. In some embodiments, the reaction mixture is heated to a temperature between 45 ℃ and 60 ℃ and then cooled to a temperature between 15 ℃ and 40 ℃.

In some embodiments, the organic phase obtained after separating the aqueous phase is subjected to one or more water washing steps (aqueous WASHING STEP), wherein after each water washing step the aqueous phase is separated, for example by one or more water washing steps with aqueous sodium chloride solution followed by separation of the aqueous phase, and subsequently by one or more water washing steps with deionized water followed by separation of the aqueous phase again. The resulting washed organic phase is then optionally distilled to reduce the water content to below 1000ppm by weight of the solution. Alternatively, in some embodiments, a small amount of water remains in the organic phase of the compound having formula (II), and subsequent coupling with the compound of formula (III) is performed in the presence of this small amount of water.

In some embodiments, the desalting reaction in step (pre 2 a) is performed on the mesylate salt (compound 1A) in a mixture of aqueous sodium hydroxide and toluene at a temperature between 45 ℃ and 60 ℃, followed by cooling the mixture to a temperature between 15 ℃ and 25 ℃, allowing the system to phase separate and separating the aqueous phase. The toluene phase obtained after separation of the aqueous phase is then optionally subjected to one or more washing steps with aqueous sodium chloride solution, followed by separation of the aqueous phase and then one or more washing steps with deionized water, followed by separation of the aqueous phase again, followed by distillation of the resulting washed toluene phase at a temperature between 50 ℃ and 65 ℃ under reduced pressure to reduce the water content to below 1000ppm by weight of the total amount of solution. Alternatively, a small amount of water remains in the toluene of the compound of formula (II), and the subsequent coupling reaction with the compound of formula (III) is carried out in the presence of this small amount of water.

As described above, in step (a), a compound of formula (II) or a salt thereof (e.g., a compound of formula (IIA) or (IIB), such as mesylate 1A) is coupled with a compound of formula (III) to provide a compound of formula (IV). In some embodiments, this process is performed in an organic solvent.

The coupling partner of formula (III) in step (a) comprises a leaving group (X ¹). It is to be understood that any convenient leaving group may be used for X ¹ in the present disclosure. In some embodiments, the leaving group (X ¹) in the compound of formula (III) is selected from halogen, carbamate, and substituted sulfonyloxy. In some embodiments, the leaving group (X ¹) in the compound of formula (III) is a sulfonyloxy group selected from methanesulfonyloxy, p-toluenesulfonyloxy, or trifluoromethanesulfonyloxy. In some embodiments, the leaving group (X ¹) is a carbamate. In some embodiments, the leaving group (X ¹) is halogen. In certain embodiments, the halogen is chloride. The coupling partner of formula (III) in step (a) also comprises a protecting group (Y ¹). The term "protecting group" refers to any group that, when bound to a functional group (e.g., a carboxylic acid moiety of a compound (including intermediates thereof)), prevents reaction at the functional group, and which can be removed by conventional chemical or enzymatic steps to reconstitute the functional group, e.g., a carboxylic acid moiety. The particular removable protecting group employed is not critical, and examples of carboxylic acid protecting groups include traditional substituents such as t-butyl, methyl, ethyl, benzyl, allyl, 1-diethylallyl, 2-trifluoroethyl, phenyl, 4-methoxybenzyl, silyl esters, orthoesters (ortho esters), esters of 2, 6-disubstituted phenols (e.g., 2, 6-dimethylphenol) and any other groups that can be chemically introduced onto a carboxylic acid group or similar functionality and subsequently selectively removed by chemical or enzymatic methods under mild conditions compatible with the nature of the product. it is to be understood that any convenient protecting group (e.g., an ester group) for the carboxylic acid moiety may be used in the present disclosure for Y ¹, and that the selection of the appropriate protecting group may be readily determined by one of skill in the art. Suitable groups for this purpose are discussed in, for example, the standard textbooks of the following chemical fields, protective Groups in Organic Synthesis [ protecting groups in organic synthesis ] by T.W.Greene and P.G.M.Wuts, 4 th edition, (John Wiley father company (John Wiley & Sons), new York, 1999), in Protecting Group Chemistry [ protecting group chemistry ] by Jeremy Robertson, 1 st edition, (oxford university press (Oxford University Press), 2000), and March' S ADVANCED Organic chemistry: reactions Mechanisms by Michael B.Smith, and Structure [ March higher organic chemistry reaction ], Mechanism and Structure ], 8 th edition, (Wiley International science Press (Wiley-INTERSCIENCE PUBLICATION), 2001). In some embodiments, the protecting group (Y ¹) is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, allyl, substituted allyl, and silyl. In some embodiments, the protecting group (Y ¹) is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2-trifluoroethyl, phenyl, 4-methoxybenzyl, 2, 6-disubstituted phenol, and silyl. In some embodiments, the protecting group (Y ¹) is tert-butyl. In some embodiments, the compound of formula (III) has the following structure 1B:

In some embodiments of the coupling reaction of step (a), the solvent is selected from toluene, t-butanol, 1, 4-dioxane, xylene, N-methyl-2-pyrrolidone, dimethylformamide, water, tetrahydrofuran, and combinations thereof. In some embodiments, the solvent is a mixture of toluene, an organic solvent, and t-butanol, an organic co-solvent.

Since the same organic solvent is used in step (prea 2) and step (a), or since solvent exchange is performed in step (prea 2), if steps (prea 1) and (prea 2) are performed before step (a), the compound of formula (II) is already present in the desired solvent. If desired, further organic solvents and, for example, organic cosolvents may be added in step (a). As will be appreciated by the person skilled in the art, during the solvent exchange of step (pre-a 2) an organic co-solvent may also be added. In some embodiments, steps (pre a 1) and (pre a 2) are performed before step (a), and the compound of formula (II) is present in toluene.

The coupling reaction in step (a) is typically a catalytic reaction. In some embodiments, the reaction is a palladium catalyzed coupling reaction performed in the presence of a base. Suitable examples of palladium catalysts are, for example, tris (dibenzylideneacetone) dipalladium and palladium (II) acetate. Suitable bases include organic bases (e.g., sodium t-butoxide and potassium t-butoxide) and inorganic bases (e.g., K ₃PO₄、K₃PO₄·H₂ O, sodium carbonate, potassium carbonate, cesium carbonate, liHMDS, naHMDS, KOH, and NaOH).

In many embodiments, anhydrous K3PO4 is used as the base. In many such embodiments, the particle size distribution is such that 90% of the particles are between less than about 140 microns and about 307 microns (including between about 140 microns and about 170 microns, including between about 160 microns and about 290 microns, and between about 180 microns and about 220 microns, and between about 200 microns and about 210 microns). In some embodiments, 90% of the particles are less than 205 microns.

In these and other embodiments, 50% of the particles are between about 35 microns and about 173 microns or less than the stated range, including between about 35 microns and about 40 microns.

In these and other embodiments, 10% of the particles are between about 7 microns and about 74 microns, including between about 7 microns and about 10 microns.

In some embodiments, the compound of formula (II) is reacted with the compound of formula (III) in a solvent (e.g., an organic solvent) in step (a) using a palladium catalyst, a base. In some embodiments, the reaction mixture further comprises a ligand.

In some embodiments, the compound of formula (IIA) or (IIB) is reacted with the compound of formula (III) in a solvent (e.g., an organic solvent) in step (a) using a palladium catalyst, a base. In some embodiments, the reaction mixture further comprises a ligand.

In some embodiments, the desalted compound of formula (II) is reacted in step (a) with the compound of formula (III) in a solvent (e.g., an organic solvent) using palladium (II) acetate, (S) -BINAP [ (S) -2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl ] or rac-BINAP as ligand. In some embodiments, (S) -BINAP is used as the ligand, and the base is selected from the group consisting of sodium tert-butoxide, potassium tert-butoxide, anhydrous K ₃PO₄、K₃PO₄·H₂ O, sodium carbonate, potassium carbonate, cesium carbonate, liHMDS, naHMDS, KOH, and NaOH.

In some embodiments, the salt of formula (IIA) or (IIB) is reacted in step (a) with the compound of formula (III) in a solvent (e.g., an organic solvent) using palladium (II) acetate, (S) -BINAP [ (S) -2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl ], (R) -BINAP [ (S) -2,2 '-bis (diphenylphosphino) -1,1' -binaphthyl ], or rac-BINAP as the ligand. In some embodiments, (S) -BINAP is used as the ligand, and the base is selected from the group consisting of sodium tert-butoxide, potassium tert-butoxide, anhydrous K ₃PO₄、K₃PO₄·H₂ O, sodium carbonate, potassium carbonate, cesium carbonate, liHMDS, naHMDS, KOH, and NaOH. In some embodiments, the salt of formula (IIA) is a mesylate salt, i.e., compound 1A.

In some embodiments, the reaction in step (a) is performed at a temperature of 70 ℃ to 80 ℃, optionally under a nitrogen atmosphere, for 2 hours or more.

In some embodiments, the compound of formula (II) or salt of formula (IIA) is reacted with the compound of formula (III) in a mixture of the organic solvent toluene and the organic co-solvent t-butanol for 2 hours or more in step (a) at a temperature between 70 ℃ and 80 ℃ using palladium (II) acetate as a catalyst, using (S) -BINAP as a ligand, and using anhydrous K ₃PO₄ or K ₃PO₄·H₂ O as a base under a nitrogen atmosphere, wherein X ¹ is Cl and Y ¹ is t-butyl.

In some embodiments, the one or more water washing steps comprise one or more washing steps with water (preferably deionized water), followed by separation of the aqueous phase, followed by one or more washing steps with aqueous HCl, followed by separation of the aqueous phase, followed by one or more washing steps with aqueous sodium chloride, followed by separation of the aqueous phase, and finally followed by one or more washing steps with deionized water again, followed by separation of the aqueous phase.

If tert-butanol is used as organic cosolvent in step (a), this organic cosolvent is removed from the organic phase during the washing step.

If step (a) is performed in an organic solvent different from the solvent used in step (b), the organic solvent used in step (a) is exchanged with the organic solvent applied in step (b) in step (a) so that the compound of formula (IV) remains in solution.

In some embodiments wherein the (organic) solvent used in step (a) is different from that in step (b), at least a portion of the (organic) solvent used in step (a) is evaporated, for example using distillation performed under reduced pressure, and the organic solvent of step (b) is added such that the compound of formula (IV) remains in solution during solvent exchange. This process may be performed by continuously evaporating the (organic) solvent used in step (a) and continuously adding the organic solvent of step (b), for example until the amount of (organic) solvent used in step (a) is below a certain threshold value, based on the total amount of solvent. Alternatively, this process may be performed batchwise in more than one step by evaporating a portion of the (organic) solvent used in step (a) and subsequently adding a portion of the organic solvent used in step (b), e.g. until the amount of (organic) solvent used in step (a) is below a certain threshold, based on the total amount of solvent.

In some embodiments, the solvent used in step (a) is a mixture of the organic solvent toluene and the organic co-solvent t-butanol. During the washing step, t-butanol is removed from the organic phase comprising the compound of formula (IV).

In some embodiments of step (a), the remaining organic solvent toluene is exchanged with acetonitrile by distilling off part of the toluene in two or more steps under reduced pressure at a temperature between 50 ℃ and 65 ℃ and adding acetonitrile in between, wherein the amount of acetonitrile added is such that a solvent mixture containing less than about 20 weight percent toluene based on the combined weight of solvents is obtained, such that the compound of formula (IV) remains in solution. In some embodiments of the compound of formula (IV), Y ¹ is tert-butyl.

Method for preparing obipratropium-step (b) from aspects (a) to (d)

In step (b) of the process for preparing compounds of formula (I) according to the present disclosure, the compound of formula (IV) is converted to the carbamate of formula (V) in an organic solvent and subsequently isolated as a solid salt of formula (VI) (wherein Y ¹ is a protecting group as described herein, for example).

In some embodiments, the organic solvent used in step (b) is selected from acetonitrile, chlorobenzene, toluene, N-methyl-2-pyrrolidone, xylene, 1, 4-dioxane, ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, dichloromethane, t-butyl methyl ether, and combinations thereof. In some embodiments, the organic solvent is acetonitrile or a mixture of chlorobenzene and dichloromethane.

As previously explained herein, the compound of formula (IV) has been provided in step (a) in the organic solvent used in step (b) either because the same organic solvent is used in step (a) as in step (b) or because a solvent exchange is performed in step (a). In some embodiments of compounds of formulas (IV), (V) and (VI), Y ¹ is tert-butyl.

In some embodiments, the organic solvent used in step (b) is a mixture of acetonitrile toluene and less than about 20 weight percent toluene based on the combined weight of the organic solvents.

In some embodiments, the conversion of the compound of formula (IV) to the corresponding carbamate of formula (V) in step (b) is performed in acetonitrile in the presence of pyridine at a temperature between 10 ℃ and 20 ℃, and less than about 20 weight percent toluene and excess ethyl chloroformate based on the combined weight of the organic solvents.

If step (b) is performed in an organic solvent different from the organic solvent used in step (c), the organic solvent used in step (b) is exchanged with the organic solvent used in step (c) in step (b) so that the compound of formula (V) remains in solution.

In some embodiments wherein the organic solvent used in step (b) is different from the organic solvent used in step (c), at least a portion of the organic solvent used in step (b) is evaporated, e.g., by distillation performed under reduced pressure, and the organic solvent of step (c) is added such that the compound of formula (V) remains in solution during the organic solvent exchange. This process may be performed by continuously evaporating the organic solvent used in step (b) and continuously adding the organic solvent of step (c), for example until the amount of organic solvent used in step (b) is below a certain threshold value based on the total amount of organic solvent. Alternatively, this process may be performed batchwise in more than one step by evaporating a portion of the organic solvent used in step (b) and subsequently adding a portion of the organic solvent used in step (c), e.g. until the amount of organic solvent used in step (b) is below a certain threshold, based on the total amount of organic solvent.

The resulting mixture is preferably treated one or more times with aqueous sodium chloride and/or HCl followed by separation of the aqueous phase and subsequently treated one or more times with aqueous bicarbonate solution followed by separation of the aqueous phase.

In some embodiments, the conversion of the compound of formula (IV) to the corresponding carbamate of formula (V) in step (b) is performed in acetonitrile with excess ethyl chloroformate at a temperature between 10 ℃ and 20 ℃ and this solvent is exchanged with isopropyl acetate by distilling off part of the acetonitrile in two or more steps under reduced pressure at a temperature of 60 ℃ or less and adding isopropyl acetate in an amount in the middle to obtain a solution of the compound of formula (V) in isopropyl acetate, wherein the solution may be treated with aqueous NaCl/HCl, followed by one or more treatments with aqueous bicarbonate, followed by separation of the aqueous phase.

Next, the compound of formula (V) dissolved in an organic solvent is converted into the corresponding salt according to formula (VI), wherein a ^n- is an anion and n is an integer from 1 to 3. The solid form of the salt according to formula (VI) is then isolated as a solid form.

In some embodiments, the salt of formula (VI) is selected from salts having An anion a ^n- selected from sulfonate (e.g., benzenesulfonate, toluenesulfonate, naphthalenesulfonate, camphorsulfonate, ethanesulfonate, ethanedisulfonate, or methanesulfonate), sulfate (e.g., methylsulfate), halogen, acetate, aspartate, benzoate, bicarbonate, acid tartrate, carbonate, citrate, decanoate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, hexanoate, hydroxynaphthoate, hydroxyethylsulfonate, lactate, lactose, malate, maleate, mandelate, lactate, nitrate, octanoate, oleate, pamoate, pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, succinate, tartrate, and theachloroate.

In some embodiments, the salt of formula (VI) is selected from salts having an anion a ^n- selected from chloride, bromide, acid tartrate, sulfate, and sulfonate.

In some embodiments, the salt of formula (VI) is selected from salts having an anion a ^n- selected from chloride, bromide, acid tartrate and methanesulfonate.

In some embodiments, the salt form of formula (VI) is a mesylate salt, including crystalline mesylate salts thereof, i.e., compound 1D:

In some embodiments of the salt of formula (VI), n is 1.

The organic solvent used in the conversion of formulas (V) to (VI) is not particularly limited, but in some embodiments is selected from the group consisting of cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, and combinations thereof. In some embodiments, isopropyl acetate or a mixture comprising methylene chloride, n-heptane and isopropanol, such as a mixture of chlorobenzene, methylene chloride, n-heptane and isopropanol, is used. It should be noted that the compounds of formula (V) have been provided in organic solvents due to the presence of solvent exchange as previously set forth herein.

Thus, in some embodiments, the organic solvent used to convert the compound of formula (V) to its corresponding salt of formula (VI) is selected from the group consisting of cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, and combinations thereof, having less than about 20 weight percent toluene and less than about 7 weight percent acetonitrile, based on the combined weight of the solvents. In some embodiments, the solvent is a mixture of isopropyl acetate, toluene, and acetonitrile, having less than about 20 weight percent toluene and less than about 7 weight percent acetonitrile, based on the combined weight of the solvents.

In some embodiments, it is preferred to add an organic co-solvent that is different from the organic solvent already used in step (b). Exemplary organic cosolvents are selected from the group consisting of cyclopentyl methyl ether, isopropyl ether, t-butyl methyl ether, ethyl acetate, isopropyl acetate, and combinations thereof, such as methyl t-butyl ether. As will be appreciated by those skilled in the art, the need and advantage of using an organic co-solvent will depend on the particular organic solvent already used in step (b). In some cases, the use of a co-solvent may be omitted.

In some embodiments, the organic solvent used to convert the compound of formula (V) to its corresponding salt of formula (VI) includes isopropyl acetate and methyl tert-butyl ether as organic co-solvents.

Subsequently, an acid is added to form a salt of formula (VI) as defined above. In some embodiments, the acid is selected from the group consisting of bitartrate, sulfuric acid, sulfonic acid, hydrogen bromide, and hydrogen chloride. In some embodiments, the acid is methanesulfonic acid. In embodiments where the salt of formula (VI) is available in crystalline form, a portion of the acid required to form the salt of formula (VI) may be added prior to crystallization and a portion added during crystallization.

The salt of formula (VI) is isolated in solid form by crystallization (if a salt of formula (VI) is available in crystalline form), filtration, one or more optional filter residue washing steps and drying.

In some embodiments, the compound of formula (V) is converted to the corresponding mesylate salt according to formula (VI) with methanesulfonic acid in an organic solvent mixture of isopropyl acetate and methyl tert-butyl ether, wherein less than about 20 weight percent toluene and less than 7 weight percent acetonitrile are present based on the combined weight of the organic solvents, followed by crystallization of the mesylate salt according to compound 1D in the organic solvent, followed by filtration, one or more optional washing steps of the filtration residue, and drying.

In some embodiments, wherein the salt according to formula (VI) is available in crystalline form, crystallization is induced by seeding with a seed crystal of the salt according to formula (VI).

In some embodiments, wherein the salt according to formula (VI) is obtainable in crystalline form, crystallizing the salt according to formula (VI) and obtaining the crystalline form of the salt according to formula (VI) is performed by adding the acid required to form the salt, stirring the resulting mixture at a temperature of 20 ℃ to 25 ℃ for more than 60 minutes, allowing crystallization to proceed with stirring for more than 120 minutes at a temperature between 15 ℃ and 25 ℃, followed by vacuum filtration of the resulting slurry, wherein the filtration residue is washed one or more times with the same organic solvent used to crystallize the salt according to formula (VI), and vacuum drying the crystalline form of the salt according to formula (VI).

In an embodiment, the invention relates to a salt according to formula (VI), wherein a ^n- is an anion, wherein n is an integer from 1 to 3. In some embodiments, the compound is a crystalline methanesulfonic acid (MSA) salt of formula (VI) (e.g., compound 1D described herein).

In some embodiments, crystallizing the mesylate of formula (VI) from an organic solvent mixture of isopropyl acetate and methyl tertiary butyl ether and obtaining the crystalline form of the mesylate according to compound 1D is performed by adding the methanesulfonic acid required to form the salt, stirring the resulting mixture at a temperature between 15 ℃ and 25 ℃ (e.g., 20 ℃) for more than 60 minutes, and then allowing crystallization to proceed while stirring at a temperature between 15 ℃ and 25 ℃ for more than 120 minutes. The resulting slurry is vacuum filtered, wherein the filter residue is washed one or more times with a mixture of isopropyl acetate and methyl tert-butyl ether and dried under vacuum to provide the crystalline form of the mesylate salt according to compound 1D.

In some embodiments, the compound of formula (VI) is obtained in a yield of at least 70% based on moles of the compound of formula (II). In some embodiments, compounds of formula (VI) are obtained at a purity of 99% or higher, for example at a purity of 99.1% or higher, 99.2% or higher, 99.3% or higher, 99.5% or higher, or even higher.

Method for preparing obipratropium-steps (c) of aspects (a) to (d)

In step (c) of the process according to the present disclosure, the isolated salt of formula (VI) or a desalted derivative thereof (e.g., a compound according to formula (V)) is alkylated with a compound of formula (VII) to provide a compound of formula (VIII):

wherein X ² is a leaving group and Y ¹ is a protecting group (e.g., as described herein).

In some embodiments of step (c), the isolated solid form of the salt according to formula (VI) (e.g., the crystalline form of the salt according to formula (VI) (e.g., crystalline mesylate, i.e., compound 1D)) is directly reacted with the compound of formula (VII) in an organic solvent to form the compound of formula (VIII) (i.e., without a desalting step).

In some embodiments of step (c), the isolated solid form of the salt according to formula (VI), e.g., the crystalline form of the salt according to formula (VI) (e.g., crystalline mesylate, i.e., compound 1D), is desalted and reacted with the compound of formula (VII) in an organic solvent to form the compound of formula (VIII). Desalting the compound of formula (VI) gives a compound according to formula (V).

When the compound of formula (VI) is subjected to a desalting step, the desalting process is performed in the same organic solvent as the subsequent reaction with the compound of formula (V). In some embodiments, the organic solvent is selected from the group consisting of xylene, n-hexane, toluene, heptane (mixtures of isomers), n-heptane, methylene chloride, chlorobenzene, and combinations thereof. In some embodiments, the organic solvent is toluene or n-heptane.

In some embodiments, step (c) is performed in the presence of a base. In some embodiments, step (c) is performed in the presence of a solid-liquid phase transfer catalyst.

In some embodiments, the base is selected from the group consisting of alkali metal hydrides, alkali metal hydroxides, alkaline earth metal hydroxides, alkali metal alkoxides, alkali metal carbonates, alkali metal bicarbonates, and amines. In some embodiments, the base is selected from alkali metal alkoxides. In some embodiments, the base is sodium tert-amylate or a mixture of sodium tert-butoxide and potassium tert-butoxide.

In some embodiments, the solid-liquid phase transfer catalyst is selected from the group consisting of t-butyl ammonium bisulfate, tetra-n-butyl ammonium bromide, tetra-n-butyl ammonium iodide, crown ethers (crown ethers), and combinations thereof. In some embodiments, the catalyst is t-butyl ammonium bisulfate.

In some embodiments, the reaction of the compound of formula (V) or (VI) with the compound of formula (VII) is performed at a temperature between 0 ℃ and 25 ℃ (e.g., 5 ℃ to 20 ℃).

The coupling partner of formula (VII) in step (c) comprises a leaving group X ². It is to be understood that any convenient leaving group may be used for X ² in the present disclosure. In some embodiments, leaving group X ² in the compound of formula (VII) is selected from halogen and substituted sulfonyloxy. In some embodiments, leaving group X ² in the compound of formula (VII) is a substituted sulfonyloxy group selected from methanesulfonyloxy, p-toluenesulfonyloxy, or trifluoromethanesulfonyloxy. In some embodiments, leaving group X ² is halogen. In certain embodiments, the halogen is bromide. In some embodiments, the compound of formula (VII) has structure 1E below.

In some embodiments, the desalting of the compound of formula (VI) and the subsequent reaction with the compound of formula (VII) in step (C) is performed in toluene as an organic solvent at a temperature of 5 ℃ to 25 ℃ in the presence of a base and a catalyst. In some embodiments, the desalting of the compound of formula (VI) and the subsequent reaction with the compound of formula (VII) in step (C) is performed in toluene as an organic solvent with stirring at a temperature between 5 ℃ and 25 ℃ in the presence of sodium tert-amyl alcohol as a base and ammonium tert-butyl bisulfate as a catalyst for about 1 to 8 hours. In some embodiments of the compound of formula (VI), Y ¹ is tert-butyl.

In some embodiments, the alkylation of the compound of formula (VI) with the compound of formula (VII) in step (C) (i.e., without an additional desalting step) is performed in toluene as an organic solvent at a temperature of 5 ℃ to 25 ℃ in the presence of a base and a catalyst. In some embodiments, alkylating the compound of formula (VI) with the compound of formula (VII) in step (C) is performed in toluene as an organic solvent with stirring at a temperature between 5 ℃ and 25 ℃ in the presence of sodium tert-amyl alcohol as a base and ammonium tert-butyl bisulfate as a catalyst for about 1 to 8 hours.

In some embodiments, step (C) comprises providing crystalline 1D, desalting the compound, and reacting the desalted compound with a compound of formula (VII) (wherein X ² is Br) at a temperature between 5 ℃ and 25 ℃ in the presence of sodium tert-amyl alcohol as a base and ammonium tert-butyl bisulfate as a catalyst in toluene as an organic solvent for about 1 to 8 hours while stirring.

In some embodiments, step (C) comprises reacting crystalline 1D with a compound of formula (VII) (wherein X ² is Br) in the presence of sodium tert-amyl alcohol as a base and ammonium tert-butyl bisulfate as a catalyst at a temperature between 5 ℃ and 25 ℃ in toluene as an organic solvent for about 1 to 8 hours while stirring.

In some embodiments of step (c), the base is the last reagent added to the reaction mixture. Without being bound by any particular theory, the inventors of the present invention have found that by adding a base as the final reagent, the number of equivalents of both the base and the compound of formula (VII) used in the reaction mixture can be reduced. Reducing the number of equivalents of the compound of formula (VII) in turn reduces the risk of bringing related impurities of formula (VII) into the final product.

Thus, step (c) results in the compound of formula (VIII) being produced in an organic solvent. In some embodiments of the compound of formula (VIII), Y ¹ is tert-butyl. In some embodiments, this reaction mixture is subjected to one or more water washing steps in step (c) to remove impurities, followed by separation of the aqueous phase and optionally one or more filtration steps to obtain a washed reaction mixture comprising the compound of formula (VIII) in an organic solvent. In some embodiments, the reaction mixture comprising the compound of formula (VIII) in the organic solvent is concentrated by distilling off a portion of the organic phase to obtain a concentrated reaction mixture comprising the compound of formula (VIII) in the organic solvent. In some embodiments, the organic solvent comprises from 30 weight percent to 40 weight percent of the compound of formula (VIII), based on the weight of the reaction mixture. In some embodiments, the organic solvent comprises 34 to 37 weight percent of the compound of formula (VIII), based on the weight of the reaction mixture.

Preferably the one or more water washing steps, the optionally performed one or more filtration steps, and the concentration step are combined, thereby obtaining a washed and concentrated reaction mixture comprising the compound of formula (VIII) in an organic solvent. In some cases, the organic solvent comprises 30 to 40 weight percent of the compound of formula (VIII). In some embodiments, the organic solvent comprises 34 to 37 weight percent of the compound of formula (VIII), based on the weight of the reaction mixture.

In some embodiments, the one or more aqueous washing steps comprise one or more washing steps with aqueous acetic acid.

In some embodiments, in step (C), the reaction mixture comprising the compound of formula (VIII) in toluene as organic solvent is subjected to one or more aqueous washing steps with aqueous acetic acid, followed by separation of the aqueous phase and subsequent distillation of part of the toluene at a temperature of typically 75 ℃ to 90 ℃ under reduced pressure to obtain a washed and concentrated reaction mixture comprising the compound of formula (VIII) in toluene, wherein the content of the compound of formula (VIII) is from 30 weight percent to 40 weight percent based on the weight of the reaction mixture. In some embodiments, the concentrated mixture comprises 34 weight percent to 37 weight percent of the compound of formula (VIII) based on the weight of the reaction mixture.

If step (c) is performed in an organic solvent different from the organic solvent used in step (d), the organic solvent used in step (c) is exchanged with the organic solvent used in step (d) in step (c) so that the compound of formula (VIII) remains in solution.

In some embodiments wherein the organic solvent used in step (c) is different from the organic solvent used in step (d), at least a portion of the organic solvent used in step (c) is preferably evaporated using distillation performed under reduced pressure, and the organic solvent of step (d) is added such that the compound of formula (VIII) remains in solution during the organic solvent exchange. This process may be performed by continuously evaporating the organic solvent used in step (c) and continuously adding the organic solvent of step (d), for example until the amount of organic solvent used in step (c) is below a certain threshold value based on the total amount of organic solvent. Alternatively, this process may be performed batchwise in more than one step by evaporating a portion of the organic solvent used in step (c) and subsequently adding a portion of the organic solvent used in step (d), e.g. until the amount of organic solvent used in step (c) is below a certain threshold, based on the total amount of organic solvent.

Process for preparing a compound of formula (I) -step (d) of aspects (a) to (d)

In step (d) of the process according to the present disclosure, the compound of formula (VIII) is converted to obpezotezole (wherein Y ¹ is a protecting group as described herein, for example) in a first organic solvent.

The choice of the first organic solvent used in step (d) is not particularly limited. In some embodiments, the first organic solvent is not an ether or an ester. In some embodiments, the first organic solvent is toluene or a mixture of n-heptane and acetic acid. As previously explained herein, the compound of formula (VIII) has been provided in step (c) in the first solvent used in step (d), either due to the use of the same organic solvent in step (c) as in step (d), or due to the solvent exchange performed in step (c).

Thus, in some embodiments, a first organic solvent as defined previously herein is provided in step (d) having from 30 to 40 weight percent of the compound of formula (VIII), e.g., from 34 to 37 weight percent of the compound of formula (VIII), based on the weight of the reaction mixture.

In some embodiments, toluene is provided in step (d) as the first organic solvent, the toluene having from 30 weight percent to 40 weight percent, e.g., from 34 weight percent to 37 weight percent, of the compound of formula (VIII), based on the weight of the reaction mixture.

Any convenient protecting group (e.g., an ester moiety) of a carboxylic acid may be used as Y ¹ in the compound of formula (VIII). The selection of the appropriate protecting groups for the carboxylic acid, as disclosed herein, can be readily determined by one of skill in the art. In some embodiments of formula (VIII), the protecting group (Y ¹) is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, allyl, substituted allyl, and silyl. In some embodiments of formula (VIII), the protecting group (Y ¹) is selected from t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2-trifluoroethyl, phenyl, 4-methoxybenzyl, 2, 6-disubstituted phenol, and silyl. In some embodiments of the compound of formula (VIII), the protecting group Y ¹ is tert-butyl. In some embodiments, converting the compound of formula (VIII) to objected is performed by contacting the compound of formula (VIII) with acetic acid (AcOH) and dry HCl in a first organic solvent (e.g., toluene or a mixture of n-heptane and acetic acid) while stirring. In some embodiments, the reaction mixture is heated to a temperature between 40 ℃ and 55 ℃ and the resulting mixture is maintained at this temperature for at least 3 hours while stirring.

The obipratropium can be isolated from the resulting mixture using techniques known to those skilled in the art.

In some embodiments, the resulting mixture comprising obsemitrapib is subjected to one or more water washing steps in step (d). In some embodiments, the one or more water washing steps in step (d) are performed as follows:

(AA) cooling the reaction mixture comprising obipratropium to a temperature between 15 ℃ and 25 ℃ and subsequently adding a mixture of n-heptane, acetonitrile and water, followed by stirring the resulting mixture at this temperature for more than 15 minutes;

(BB) allowing the system phase obtained in step (AA) to separate into an organic phase and an aqueous phase, and separating said two phases;

(CC) adding a mixture of n-heptane, acetonitrile, toluene and water to the aqueous phase obtained in step (BB), followed by stirring the resulting system at a temperature between 15 ℃ and 25 ℃ for more than 15 minutes;

(DD) allowing the system phase obtained in step (CC) to separate into an organic phase and an aqueous phase, and separating said two phases;

(EE) combining the organic phase obtained in step (BB) with the organic phase obtained in step (DD), adding water, and stirring the resulting system at a temperature between 15 ℃ and 25 ℃ for more than 15 minutes;

(FF) allowing the system phase obtained in step (EE) to separate into an organic phase and an aqueous phase, and separating said two phases;

(GG) adding water to the organic phase obtained in step (FF) and stirring the resulting system at a temperature between 15 ℃ and 25 ℃ for more than 15 minutes;

(HH) allowing the system phase obtained in step (GG) to separate into an organic phase and an aqueous phase, and separating the two phases;

(II) adding an aqueous solution of sodium citrate tribasic dihydrate to the organic phase obtained in step (HH), followed by stirring the resulting mixture at a temperature between 15 ℃ and 25 ℃ for more than 15 minutes;

(JJ) allowing the system phase obtained in step (II) to separate into an organic phase and an aqueous phase, and separating the two phases;

(KK) adding water to the organic phase obtained in step (JJ) and stirring the resulting system at a temperature between 15℃and 25℃for more than 15 minutes, and

(LL) allowing the system phase obtained in step (KK) to separate into an organic phase and an aqueous phase, and separating the two phases.

Steps (AA) to (LL) in this example yield the washed compound of formula (I) in an organic solvent mixture comprising n-heptane, acetonitrile and a first organic solvent. In some embodiments, the first solvent is toluene.

In some embodiments, wherein the first organic solvent has not been composed primarily of cyclopentyl methyl ether, the organic solvent mixture is exchanged with CPME in a subsequent step (MM) such that the obipratropium remains in solution.

Thus, in some embodiments, step (MM) is performed after step (LL), wherein at least a portion of the solvent in the organic solvent mixture obtained in step (LL) is evaporated, e.g., by distillation under reduced pressure, and wherein cyclopentyl methyl ether is added such that the obipratropium remains in solution during solvent exchange. In some embodiments, the process forms a solution of obrisetrapezin in cyclopentyl methyl ether at a concentration of between 30 weight percent and 40 weight percent based on the weight of the solution. In some embodiments, the concentration of obrisedronate in the cyclopentyl methyl ether is from 33 weight percent to 37 weight percent based on the weight of the solution, less than 1 weight percent of the first organic solvent is present, based on the weight of the solution, and less than 1 weight percent of n-heptane.

This process may be performed by continuously evaporating the solvent in the organic solvent mixture obtained in step (LL) and by continuously adding cyclopentyl methyl ether, for example, until the amount of the specific solvent in the organic solvent mixture is below a certain threshold value, based on the total amount of organic solvents. Alternatively, this process may be performed batchwise in more than one step by evaporating part of the solvent in the organic solvent mixture obtained in step (LL) and subsequently adding cyclopentyl methyl ether, for example until the amount of the specific solvent in the organic solvent mixture, based on the total amount of solvents, is below a certain threshold.

In some embodiments, the first organic solvent is toluene, step (MM) is performed after step (LL), wherein at least a portion of the n-heptane, acetonitrile and toluene in the organic solvent mixture obtained in step (LL) is evaporated, for example by distillation at a temperature of 45 ℃ or less and under reduced pressure (in vacuo), with the addition of cyclopentyl methyl ether in between, such that the obipsilate remains in solution during solvent exchange, thereby obtaining a solution of obipsilate in cyclopentyl methyl ether at a concentration between 30 and 40 weight percent. In some embodiments, the concentration of obrisedronate in the cyclopentyl methyl ether is from 33 weight percent to 37 weight percent based on the weight of the solution having less than 0.5 weight percent toluene, less than 0.5 weight percent acetonitrile, and less than 2.7 weight percent n-heptane.

Method for preparing crystalline obiprapib HCl-steps (e) to (f) except for aspects (a) to (d)

In some embodiments of the subject methods, steps (e) through (f) are performed after step (d), wherein the obipratropium is treated with HCl, for example, in a suitable solvent. Such solvents may be aqueous or organic. In some embodiments, the use of an organic solvent provides crystalline obiprift HCl.

In some embodiments, the organic solvent used in step (e) comprises a mixture of solvent and anti-solvent. In some embodiments, the solvent is selected from the group consisting of methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl tert-butyl ether, cyclopentyl methyl ether, N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1, 4-dioxane, 1, 2-difluorobenzene, toluene, hexafluoroisopropanol, and water. In some embodiments, the antisolvent is selected from the group consisting of n-heptane, n-hexane, n-pentane, and cyclohexane. In some embodiments, HCl has sufficient solubility in the anti-solvent such that it can be used as a suitable solvent. In some embodiments, the organic solvent used in step (e) comprises a mixture of cyclopentyl methyl ether and n-heptane. In some embodiments, the organic solvent used in step (e) further comprises toluene.

In some embodiments, step (e) comprises providing obetatrapib in a mixture of cyclopentyl methyl ether and n-heptane, increasing the temperature to between 35 ℃ and 40 ℃ while stirring, adding dry HCl to the cyclopentyl methyl ether, and increasing the temperature again to between 50 ℃ and 55 ℃, then adding additional n-heptane as an anti-solvent. At this point, a small portion of the reaction mixture may be extracted and cooled to a temperature between 10 ℃ and 15 ℃ to obtain a slurry of crystals of crystalline obrisedronate HCl in a mixture of cyclopentyl methyl ether and n-heptane (referred to herein as a "seed slurry"). Optionally, all or a portion of the seed slurry of crystalline olanzapine HCl may then be seeded back into the reaction mixture. The seed aids in nucleation, but is not required, and thus the process described herein may be performed without seeding (seeding). The resulting reaction mixture is then cooled to a temperature between 5 ℃ and 15 ℃ (e.g., 10 ℃ to 15 ℃) and then the crystalline obiprift HCl is crystallized from the system while stirring. In some embodiments, the crystalline obiprap HCl is crystallized over a period of 12 hours or more, then filtered (e.g., by a filter dryer), subjected to one or more optional washing steps (e.g., washing with a mixture of cyclopentyl methyl ether and n-heptane), and dried. In some cases, the wet cake of crystalline olanzapine HCl is vacuum dried in multiple steps using temperatures of 25 ℃ to 30 ℃,30 ℃ to 40 ℃, 40 ℃ to 50 ℃, then 50 ℃ to 55 ℃, e.g., 25 ℃, 35 ℃, 46 ℃ and 54 ℃.

Thus, in some embodiments, the method of preparing crystalline obiprift HCl includes adding seed crystals (e.g., as a seed slurry). The seed crystals of crystalline obiprift HCl may be formed into a slurry by following step (i) set forth above and, after addition of dry HCl to cyclopentyl methyl ether and anti-solvent n-heptane, extracting a small portion of the reaction mixture, cooling it to a temperature between 10 ℃ and 15 ℃, thereby providing a slurry of crystals of crystalline obiprift HCl in cyclopentyl methyl ether and n-heptane.

In some embodiments, the organic solvent used in step (e) comprises a mixture of cyclopentyl methyl ether and n-heptane. Thus, in one embodiment, step (e) comprises providing obetatrapib in a mixture of cyclopentyl methyl ether and n-heptane, increasing the temperature to a temperature of from 35 ℃ to 45 ℃ while stirring, adding dry HCl to the cyclopentyl methyl ether and increasing the temperature again to 50 ℃ to 55 ℃, adding additional n-heptane as an anti-solvent, optionally adding seed crystals of crystalline obetatrapib HCl (e.g., as a seed slurry prepared as described herein), cooling to a temperature between 5 ℃ and 15 ℃ (e.g., from 10 ℃ to 15 ℃) and then crystallizing crystalline obetatrapib HCl from the system while stirring. In some embodiments, the crystalline obiprift HCl is crystallized over a period of at least 12 hours, followed by filtration, one or more optional washing steps (e.g., washing with a mixture of cyclopentyl methyl ether and n-heptane), and drying. In some embodiments, the crystalline obiprift HCl is dried in vacuo. In some embodiments, the crystalline obiprift HCl is dried in a vacuum oven at 25mbar pressure and a temperature of 55 ℃ for 10 hours or more. In some embodiments, after the drying procedure, the crystalline obiprift HCl includes less than 0.1 weight percent residual cyclopentyl methyl ether.

In some embodiments previously described herein, step (MM) of step (d) forms a solution of objected to be used in step (d) comprising less than 1 weight percent of the first organic solvent and less than 1 weight percent of n-heptane in the cyclopentyl methyl ether at a concentration of between 30 weight percent and 40 weight percent, e.g., 33 weight percent to 37 weight percent, based on the weight of the solution. In some embodiments previously described herein, step (MM) of step (d) forms a solution of objected to be mixed with the cyclopentyl methyl ether, the solution comprising less than 1 weight percent toluene and less than 1 weight percent n-heptane, at a concentration of between 30 weight percent and 40 weight percent, e.g., 33 weight percent to 37 weight percent, based on the weight of the solution. These solutions can advantageously be used in step (e) after the addition of n-heptane. N-heptane may also be added in step (d), as will be appreciated by those skilled in the art.

Thus, in some embodiments, step (e) comprises forming a solution of obetatrapib in cyclopentyl methyl ether at a concentration of between 30 and 40 weight percent, e.g., 33 to 37 weight percent, based on the weight of the solution, the solution comprising less than 1 weight percent of the first organic solvent used in step (d) (e.g., toluene) and less than 1 weight percent of n-heptane, adding n-heptane, increasing the temperature to 35 to 45 ℃ while stirring, adding dry HCl in cyclopentyl methyl ether and increasing the temperature again to 50 to 55 ℃, adding additional n-heptane as an antisolvent, optionally adding seeds of crystalline obetatrapib HCl (e.g., as seed slurry as prepared herein), cooling to a temperature of between 5 and 15 ℃ (e.g., 10 to 15 ℃), followed by crystallizing crystalline obetaxel from the system while stirring, e.g., during a period of at least 12 hours, then performing filtration with n-methyl ether, and drying the mixture, or more than one step. In some cases, the wet cake of crystalline olanzapine HCl is vacuum dried in multiple steps using temperatures of 25 ℃ to 30 ℃, 30 ℃ to 40 ℃, 40 ℃ to 50 ℃, then 50 ℃ to 55 ℃, e.g., 25 ℃, 35 ℃, 46 ℃ and 54 ℃.

In some embodiments, step (f) comprises the steps of:

(aa) providing crystalline obiprift HCl;

(bb) dissolving the crystalline olanzapine HCl in ethanol while stirring. In some embodiments, the temperature is between 15 ℃ and 25 ℃;

(cc) adding an aqueous NaOH solution to the solution obtained in step (bb) and stirring the resulting mixture for at least 4 hours, for example at a temperature of 20 ℃ to 25 ℃, to obtain a solution of the sodium salt of obsemitrapib;

(dd) optionally filtering the solution obtained in step (cc);

(ee) preparing a CaCl2 solution by adding deionized water to CaCl2 while stirring, followed by adding ethyl acetate as a co-solvent, and stirring the resulting mixture for 10 minutes to 30 minutes;

(ff) cooling the CaCl2 solution obtained in step (ee) to a temperature between 8 ℃ and 12 ℃ and adding to the solution obtained in step (dd) (or (cc)) via a filter while stirring at said temperature;

(gg) stirring the slurry from step (ff) for about 1 hour to about 10 hours. In some embodiments, the slurry is stirred at a temperature between 8 ℃ and 12 ℃;

(hh) separating solids from the slurry obtained in step (gg) by filtration. In some embodiments, the separation is performed at a temperature between 8 ℃ and 12 ℃;

(ii) The filter residue obtained in step (hh) is washed with water in one or more washing steps. In some embodiments, the washing is performed at a temperature between 8 ℃ and 12 ℃, and

(Jj) drying the washed residue obtained in step (ii) in vacuo, for example at a temperature between 40 ℃ and 50 ℃ for more than 16 hours (e.g. 200 hours or more), to obtain amorphous obiprifleside calcium.

In some embodiments of the subject process, crystalline objected HCl is isolated in step (f) with a purity of 98% or greater, such as 98.5% or greater, 99% or greater, 99.5% or greater, or even greater.

Yet another embodiment of the present disclosure relates to crystalline obiprift HCl.

In some embodiments, crystalline obiprift HCl is stored at controlled room temperature and under nitrogen atmosphere and protected from moisture to prevent the formation of amorphous solids because of the hygroscopicity of crystalline obiprift HCl including crystalline obiprift HCl.

Method for preparing amorphous obiprift calcium-steps (g) to (h) except for aspects (a) to (f)

In some embodiments of the subject process, steps (g) through (h) are performed after step (f), in which steps (g) through (h) crystalline olanzapine HCl is converted to amorphous olanzapine calcium hemi- (formula IB):

In some embodiments of step (g), the preparation of amorphous obiprift hemi-calcium comprises steps (g 1) to (g 3) as follows:

(g1) Converting the crystalline obiprifepin HCl of step (f) to obiprifepin in an organic solvent;

(g2) Treating the obipratropium in an organic solvent with aqueous sodium hydroxide to form the sodium salt of obipratropium, and

(G3) Treating the sodium salt of obipratropium with an aqueous solution of calcium chloride to form amorphous obipratropium calcium chloride;

wherein the compounds in steps (g 1) and (g 2) are not isolated.

Thus, in some embodiments, step (g 1) comprises the steps of:

(aa) providing crystalline obiprapib HCl as defined or obtained in step (f);

(Dd) performing two or more distillations on the washed organic phase resulting from step (cc) at a temperature of 50 ℃ or less (e.g., 30 ℃ or less), with the intermediate addition of ethanol, thereby obtaining a solution of the compound of obichrexed in ethanol. In some embodiments, step (g 2) comprises the steps of:

(Ff) optionally filtering the solution obtained in step (ee).

In some embodiments, step (g 3) comprises the steps of:

(gg) preparing a CaCl2 solution by adding deionized water to CaCl2 while stirring, followed by adding ethyl acetate as a co-solvent, and stirring the resulting mixture for 10 minutes to 30 minutes;

(hh) cooling the CaCl2 solution obtained in step (gg) to a temperature of 8 ℃ to 12 ℃ and adding to the solution obtained in step (ff) or (ee) via a filter while stirring at said temperature;

(ii) Stirring the slurry from step (hh) for about 1 to 10 hours. In some embodiments of step (ii), the stirring is performed at a temperature between 8 ℃ and 12 ℃;

In some embodiments, step (g) comprises the steps of:

(aa) providing crystalline obiprapib HCl as defined or obtained in step (f);

(dd) optionally filtering the solution obtained in step (cc);

(ff) cooling the CaCl2 solution obtained in step (ee) to a temperature between 8 ℃ and 12 ℃ and adding to the solution obtained in step (dd) or (cc) via a filter while stirring at said temperature;

(gg) stirring the slurry from step (ff) for about 1 to 10 hours. In some embodiments, the slurry is stirred at a temperature between 8 ℃ and 12 ℃;

(Jj) drying the washed residue obtained in step (ii) in vacuo, for example at a temperature between 40 ℃ and 50 ℃ for more than 16 hours (e.g. 50 hours, 100 hours, 150 hours or 200 hours, or even more), to obtain the amorphous hemicalcium salt of formula (IB).

In some embodiments, amorphous obiprift calcium is stored hermetically and protected from light at temperatures below 30 ℃.

In many embodiments of the present disclosure, amorphous obiprift calcium is processed to achieve a particle size distribution. In many embodiments, such processing is by grinding. Examples of milling include hammer milling, ball milling, and jet milling (jet milling). In other embodiments, spray drying may be used to achieve the particle size distribution. Thus, in some embodiments of the present disclosure, spray-dried amorphous obiprift calcium is provided. Examples of spray milled amorphous obipratropium calcium are provided in example 11.14.

In many embodiments of the present disclosure, non-milled amorphous obiprift calcium is provided. In many embodiments of the present disclosure, milled amorphous obipratropium calcium is provided.

In many embodiments, the particle size distribution of amorphous obiprift calcium is such that 90% of the particles have a diameter of about 15 microns or less. In these and other embodiments, 90% of the particles have a diameter of about 14 microns or less, 13 microns or less, 12 microns or less, 11 microns or less, 10 microns or less, 9 microns or less, 8 microns or less, 7 microns or less, 6 microns or less, 5 microns or less, 4 microns or less, or 3 microns or less.

In some embodiments, 90% of the particles have a diameter between about 6 microns and 15 microns.

In these and other embodiments, the particle size distribution of amorphous obiprift halflic calcium is such that 50% of the particles have a diameter of about 5 microns or less, such as, for example, 4 microns or less or 3 microns or less.

In these and other embodiments, the particle size distribution of amorphous obiprift halflic calcium is such that 10% of the particles have a diameter of about 2 microns or less.

According to the process of the present disclosure, amorphous obipratropium calcium of the present disclosure can be made with high chemical purity. Such purity levels include purities of greater than 98.0%, such as greater than 98.1％、98.2％、98.3％、98.4％、98.5％、98.6％、98.7％、98.8％、98.9％、99.0％、99.1％、99.2％、99.3％、99.4％、99.5％、99.6％、99.7％、99.8％ or 99.9% or greater. The highest level of purity, e.g. greater than 99.8% or 99.9% purity, is more easily achieved by a process in which crystalline obiprap HCl is used as an intermediate.

As noted above, also provided herein is an amorphous calcium salt of obipratropium, including amorphous obipratropium calcium half. Novel intermediates for the synthesis of olanzapine and salts of olanzapine are also provided.

Accordingly, the subject methods have been described with reference to certain embodiments discussed above. It will be appreciated that various modifications and alternatives to those embodiments will be apparent to those skilled in the art.

In certain preferred embodiments of the present invention, the olanzapine included in the pharmaceutical composition of the present invention, in the method of the present invention, included in a unit dosage form (included in a pharmaceutical kit), etc., is in the form of a salt of olanzapine, more particularly a salt described by one or more of the following non-limiting clauses:

Clause 1. An amorphous calcium salt of obipratropium.

Clause 2. Amorphous obiprifflic calcium.

Clause 3. Stable amorphous obiprift hemi-calcium.

Clause 4. Substantially pure amorphous obiprift hemi-calcium.

Clause 5 the amorphous obiprift hemi-calcium salt of clauses 2-4 that is substantially free of any crystalline salt of obiprift hemi-calcium.

Clause 6. Amorphous obiprift hemi-calcium according to clauses 2 to 5 having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern of figure 49.

Clause 7 the amorphous obiprift calcium of clauses 2-5 having an X-ray powder diffraction pattern comprising one or more X-ray powder diffraction peaks at about 3.4 ° 2Θ, about 7.0 ° 2Θ, and about 9.2 ° 2Θ.

Clause 8. The amorphous obiprift calcium of clauses 2-7, wherein the amorphous obiprift calcium does not undergo birefringence.

Clause 9 the amorphous obiprift calcium of clauses 2-8 having a glass transition temperature at a value between about 107 ℃ and about 112 ℃.

Clause 10. Amorphous obiprift hemi-calcium of clause 9, wherein the glass transition temperature is measured using modulated differential scanning calorimetry.

Clause 11. Amorphous obiprift hemi-calcium of clause 10, wherein the measurement using modulated differential scanning calorimetry uses an open sample pan vessel.

Clause 12 the amorphous obiprift hemi-calcium of clause 11, wherein the opening is a pinhole.

Clause 13 the amorphous obiprift calcium of clauses 8-12, wherein the glass transition temperature is at a value between about 110 ℃ and about 112 ℃.

Clause 14. Amorphous obiprift hemi-calcium of clauses 2-13 having a glass transition temperature of less than about 100 ℃ when measured by differential scanning calorimetry using a closed sample pan vessel.

Clause 15 the amorphous obiprift hemi-calcium of clause 14, having a glass transition temperature at a value between about 70 ℃ and about 92 ℃ when measured by differential scanning calorimetry using a closed sample pan vessel.

Clause 16 the amorphous obiprift calcium of clauses 2-15, having a weight loss of less than about 1% when heated to about 200 ℃.

Clause 17 the amorphous obiprift hemi-calcium of clause 16, wherein the weight loss is between about 0.8% and about 0.95%.

Clause 18 the amorphous obiprift hemi-calcium of clause 17, wherein the weight loss is between about 0.84% and about 0.92%.

Clause 19 the amorphous obiprift calcium of clauses 2-18, having a water content of less than about 5%.

Clause 20 the amorphous obiprift hemi-calcium of clause 19 having a water content of less than about 4%.

Clause 21 the amorphous obiprift hemi-calcium of clause 20 having a water content of less than about 3%.

Clause 22 the amorphous obiprift hemi-calcium of clause 19 having a water content between about 0.5% and about 1.5%.

Clause 23 the amorphous obiprift calcium of clauses 2-22, in bulk form or formulated composition, having a particle size distribution wherein about 90% of the particles have a diameter of about 15 microns or less.

Clause 24 the amorphous obiprift hemi-calcium of clause 23, wherein about 90% of the particles have a diameter between about 6 microns and about 15 microns.

Clause 25 the amorphous obiprift calcium of clause 24, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 14 microns or less.

Clause 26 the amorphous obiprift calcium of clause 25, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 13 microns or less.

Clause 27 the amorphous obiprift calcium of clause 26, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 12 microns or less.

Clause 28 the amorphous obiprift calcium of clause 27, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 11 microns or less.

Clause 29 the amorphous obiprift calcium of clause 28, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 10 microns or less.

Clause 30 the amorphous obiprift calcium of clause 29, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 9 microns or less.

Clause 31 the amorphous obiprift calcium of clause 30, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 8 microns or less.

Clause 32 the amorphous obiprift calcium of clause 31, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 7 microns or less.

Clause 33 the amorphous obiprift calcium of clause 32, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 6 microns or less.

Clause 34 the amorphous obiprift calcium of clause 33, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 5 microns or less.

Clause 35 the amorphous obiprift calcium of clause 34, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 4 microns or less.

Clause 36 the amorphous obiprift calcium of clause 35, having a particle size distribution wherein about 90% or more of the particles have a diameter of about 3 microns or less.

Clause 37 the amorphous obiprift calcium of clauses 2-36, in bulk form or formulated composition form, having a particle size distribution wherein about 50% of the particles have a diameter of about 5 microns or less.

Clause 38 the amorphous obiprift hemi-calcium of clause 37 having a particle size distribution wherein about 50% of the particles have a diameter of about 4 microns or less.

Clause 39 the amorphous obiprift calcium of clause 38, having a particle size distribution wherein about 50% of the particles have a diameter of about 3 microns or less.

Clause 40 the amorphous obiprift calcium of clauses 2-39, in bulk form or formulated composition form, having a particle size distribution wherein about 10% of the particles have a diameter of about 2 microns or less.

Clause 41 the amorphous obiprift calcium of clauses 2-40 having a chemical purity of at least 98.0%.

Clause 42 the amorphous obiprift hemi-calcium of clause 41 having a chemical purity of at least 99.0%.

Clause 43 the amorphous obiprift hemi-calcium of clause 42 having a chemical purity of at least 99.5%.

Clause 44 the amorphous obiprift hemi-calcium of clause 43 having a chemical purity of at least 99.6%.

Clause 45 the amorphous obiprift hemi-calcium of clause 44 having a chemical purity of at least 99.7%.

Clause 46 the amorphous obiprift hemi-calcium of clause 45 having a chemical purity of at least 99.8%.

Clause 47 the amorphous obiprift hemi-calcium of clause 46 having a chemical purity of at least 99.9%.

Clause 48 the amorphous obiprift semi-calcium of clauses 2-47 having a solid state ¹³ C-NMR spectrum substantially the same as the solid state ¹³ C-NMR spectrum of fig. 65.

Clause 49 the amorphous obiprift hemi-calcium of clauses 2-48 having a solid state ¹³ C-NMR spectrum wherein no peak is present at about 22.1 ppm.

Clause 50 the amorphous obiprift hemi-calcium of clauses 2-49 having a solid state ¹³ C-NMR spectrum wherein no peak is present at about 29.5 ppm.

Clause 51. Amorphous, unground, obiprift hemi-calcium.

Clause 52. A ground amorphous obiprift hemi-calcium.

Clause 53 the amorphous obiprift calcium of clauses 2-50, wherein the amorphous obiprift calcium has been ground.

Clause 54 the amorphous obiprift calcium of clauses 2-50 or 53, wherein the amorphous obiprift calcium has been jet milled.

Clause 55 the amorphous obiprift calcium of clauses 2-50 or 53-54, wherein the amorphous obiprift calcium has been spray dried.

Clause 56 an amorphous calcium obiprift-hemiate prepared by a synthetic process, wherein an intermediate in the process comprises crystalline obiprift-HCl.

Clause 57 the amorphous obiprift calcium of clauses 2-56, wherein the amorphous obiprift calcium is prepared by a synthesis process, wherein an intermediate in the process comprises crystalline obiprift HCl.

Clause 58 is an obiprift HCl.

Clause 59, a crystalline obiprift HCl.

Clause 60. An amorphous HCl obiprapib compound.

Clause 61. Solvates of HCl obiprifepin as recited in clauses 58-60.

Clause 62. The olanzapine HCl of clauses 58-61, wherein the weight percentage of HCl is between about 0.01% and about 8%.

Clause 63 a composition comprising crystalline obiprift HCl of any one of clauses 58 to 62.

Clause 64 the crystalline obiprift HCl of clauses 58-60 or 62-63, wherein the crystalline obiprift HCl is a solvate.

Clause 65 the crystalline obiprift HCl of clause 64, wherein the solvate comprises obiprift and hydrochloric acid.

The crystalline obiprift HCl of clause 66, wherein the solvate comprises an organic solvent.

Clause 67. Crystalline olanzapine HCl of clause 66, wherein the solvate comprises a solvent in which the solubility is sufficient to dissolve enough HCl to deliver enough HCl to produce crystalline olanzapine HCl.

Clause 68 the solvate of any of clauses 61 or 64 to 67, wherein the solvent of the solvate is selected from the group consisting of methanol, ethanol, isopropanol, acetic acid, acetonitrile, acetone, methyl isobutyl ketone, isopropyl acetate, tetrahydrofuran, methyl tert-butyl ether, cyclopentyl methyl ether (CPME), N-methyl-2-pyrrolidone, dimethyl sulfoxide, dimethylformamide, 2-methyl-tetrahydrofuran, dichloromethane, 1, 4-dioxane, 1, 2-difluorobenzene, toluene, and hexafluoroisopropanol.

Clause 69 the crystalline obiprift HCl of clause 68, wherein the solvent is CPME.

Clause 70 the crystalline obiprift HCl of any one of clauses 58 to 59 or 61 to 69, having an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern in fig. 67.

Clause 71 the crystalline obiprift HCl of any one of clauses 58 to 59 or 61 to 69, having an X-ray powder diffraction pattern comprising peaks at about 9.8°2Θ.

Clause 72 the crystalline obsemitrapi HCl of any one of clauses 58-59, 61-69, or 71, having an X-ray powder diffraction pattern comprising one or more peaks at about 8.1°2Θ, about 9.8°2Θ, about 13.8°2Θ, about 16.7°2Θ, and about 19.5°2Θ.

Clause 73. A salt according to formula (VI):

wherein Y ¹ is a protecting group, an-is An anion, and n is An integer from 1 to 3.

Clause 74 the salt of clause 73, wherein the compound is a mesylate salt having the structure:

Clause 75 the crystalline mesylate salt of compound 1D of clause 74.

Clause 76 the crystalline mesylate salt of compound 1D of clause 75, having a powder diffraction pattern substantially the same as any of the four X-ray powder patterns shown in figure 68.

Clause 77 the crystalline mesylate salt of compound 1D of clause 75, having an X-ray powder diffraction pattern comprising one or more peaks at about 5.2 degrees 2Θ and about 9.1 degrees 2Θ.

Clause 78 the crystalline mesylate salt of compound 1D of clauses 75-77, having an X-ray powder diffraction pattern comprising one or more peaks at about 9.1°2Θ, about 15.9°2Θ, about 16.5°2Θ, about 17.2°2Θ, about 18.6°2Θ, and about 19.2°2Θ.

In certain preferred embodiments of the present invention, the olanzapine included in the pharmaceutical composition of the present invention, in the method of the present invention, included in a unit dosage form (included in a pharmaceutical kit), etc., is in the form of a salt of olanzapine, more particularly a salt prepared by a process described in one or more of the following non-limiting clauses may be used:

clause 79 a method of preparing obipratropium, wherein the method comprises:

(a) Preparing a compound of formula (IV) by coupling a compound of formula (II) or a salt thereof with a compound of formula (III):

wherein X ¹ is a leaving group and Y ¹ is a protecting group;

Wherein Y ¹ is a protecting group, an-is An anion, and n is An integer from 1 to 3;

Wherein X ² is a leaving group and Y ¹ is a protecting group, and

(D) Converting the compound of formula (VIII) to obiprifepin, wherein the reaction steps (a) to (d) are performed in an organic solvent, optionally without isolation of compounds (IV), (V) and (VIII) from the organic solvent, and wherein the process does not require chromatography.

Clause 80. The method of clause 79, wherein the compound of formula (II) in step (a) is obtained by applying the following steps prior to step (a):

(pre-a 1) providing a compound of formula (IIA) or (IIB):

(pre a 2) desalting the compound of formula (IIA) or (IIB) to obtain the compound of formula (II);

wherein the reaction in step (pre a 2) is performed in an organic solvent and the compound of formula (II) is optionally not isolated in an organic solvent and the process does not require chromatography.

The method of clause 81, wherein the salt of formula (IIA) or (IIB) is selected from salts having an anion Am-selected from sulfonate, sulfate, halogen, acetate, aspartate, benzoate, bicarbonate, acid tartrate, carbonate, citrate, decanoate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, hexanoate, hydroxynaphthoate, hydroxyethylsulfonate, lactate, malate, maleate, mandelate, lactate, nitrate, octanoate, oleate, pamoate, pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, succinate, tartrate, and theachloroate, wherein sulfonate can be benzenesulfonate, toluenesulfonate, naphthalenesulfonate, camphorsulfonate, ethanesulfonate, ethanedisulfonate, or methanesulfonate, sulfate can be methyl sulfate, and halogen can be chloride, iodide, or bromide.

Clause 82. The method of clause 81, wherein the salt having the anion a ^m- is selected from the group consisting of chloride, bromide, acid tartrate, sulfate, and sulfonate.

Clause 83 the method of clause 82, wherein the salt having the anion a ^m- is selected from the group consisting of chloride, bromide, acid tartrate, and mesylate.

The method of any one of clauses 79 to 83, wherein Y ¹ in the compound of formulas (III) to (VI) and (VIII) is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, allyl, substituted allyl, and silyl.

The method of clause 85, wherein Y ¹ in the compounds of formulas (III) to (VI) and (VIII) is selected from the group consisting of t-butyl, methyl, ethyl, benzyl, allyl, substituted allyl, 2-trifluoroethyl, phenyl, 4-methoxybenzyl, 2, 6-disubstituted phenol, and silyl.

The method of clause 86, 85, wherein Y ¹ in the compounds of formulas (III) to (VI) and (VIII) is t-butyl.

The method of any of clauses 79 to 86, wherein the salt of formula (VI) is selected from salts having an anion a ^n-, the anion a ^n- is selected from sulfonate, sulfate, halogen, acetate, aspartate, benzoate, bicarbonate, acid tartrate, carbonate, citrate, decanoate, fumarate, glucoheptonate, gluconate, glutamate, glycolate, hexoate, hydroxynaphthoate, hydroxyethylsulfonate, lactate, malate, maleate, mandelate, lactate, nitrate, octanoate, oleate, pamoate, pantothenate, phosphate, polygalacturonate, propionate, salicylate, stearate, succinate, tartrate, and theachlorate, wherein sulfonate can be benzenesulfonate, toluenesulfonate, naphthalenesulfonate, camphorsulfonate, ethanesulfonate, ethanedisulfonate, or methanesulfonate, sulfate can be methylsulfate, and halogen can be chloride, iodide, or bromide.

The method of clause 88, wherein the salt having the anion a ^n- is selected from the group consisting of chloride, bromide, acid tartrate, sulfate, and sulfonate.

Clause 89 the method of clause 87, wherein the salt having the anion a ^n- is selected from the group consisting of chloride, bromide, acid tartrate, and mesylate.

The method of clause 90, 87, wherein the salt form of formula (VI) is a mesylate salt, compound 1D:

clause 91. The method of clause 90, wherein the mesylate salt is crystalline.

The method of any one of clauses 79 to 91, wherein X ¹ in the compound of formula (III) is selected from the group consisting of halogen, carbamate, and substituted sulfonyloxy.

The method of clause 93, 92, wherein X ¹ in the compound of formula (III) is halogen.

The method of clause 94, clause 93, wherein the halogen is chloride.

The method of any one of clauses 79 to 94, wherein X ² in the compound of formula (VII) is selected from halogen and substituted sulfonyloxy.

The method of clause 96, 95, wherein X ² in the compound of formula (III) is halogen.

Clause 97. The method of clause 96, wherein the halogen is bromide.

Clause 98 a process for preparing amorphous hemicalcium salt of obrisetrapib, wherein the process comprises:

(i) Treating the obipratropium with HCl to obtain a crystalline obipratropium HCl compound;

(ii) Separating the crystalline obiprifepin HCl compound;

(iii) Preparing an amorphous hemicalcium salt of olanzapine from the crystalline olanzapine HCl compound separated in step (ii), and

(Iv) Separating the amorphous hemicalcium salt of the obipratropium.

The method of clause 99, clause 98, wherein the crystalline obiprift HCl compound isolated in step (ii) comprises a compound of formula (IH):

Where y varies from 0.002 to 1.5.

The method of clause 98 or 99, wherein the preparation of the amorphous hemicalcium salt of formula (I) in step (iii) comprises the steps of:

(iii-1) converting the crystalline obiprift HCl compound of step (ii) to provide obiprift in one or more suitable solvents selected from the group consisting of organic solvents and aqueous solvents;

(Iii-3) treating the sodium salt of obipratropium with an aqueous solution of calcium chloride to form an amorphous hemicalcium salt of obipratropium;

wherein the compounds in steps (iii-1) and (iii-2) are optionally not isolated.

The method of any one of clauses 98 to 100, wherein the amorphous hemicalcium salt of obipratropium is amorphous obipratropium hemicalcium.

The method of any one of clauses 98 to 101, wherein the amorphous calcium salt of obipratropium is isolated in a chemical purity of at least 99%.

Clause 103 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.1%.

Clause 104 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.2%.

Clause 105 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.3%.

Clause 106 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.4%.

Clause 107 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.5%.

Clause 108 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.6%.

Clause 109 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.7%.

Clause 110 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.8%.

Clause 111 the method of clause 102, wherein the amorphous calcium salt of obipratropium is isolated in a purity of at least 99.9%.

The method of any one of clauses 102-111, wherein the amorphous calcium salt of obipratropium is amorphous obipratropium calcium half-hydrate.

Clause 113 a pharmaceutical composition comprising the amorphous salt of obiprift calcium of any one of clauses 1 to 57, and one or more pharmaceutically acceptable carriers.

Clause 114 the pharmaceutical composition of clause 113, wherein the amorphous salt of obiprifflic calcium is amorphous obiprifflic calcium.

Clause 115. A method of treating a subject having a cardiovascular disease or having an increased risk of having a cardiovascular disease, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of clause 113 or 114.

Clause 116 an amorphous calcium salt of obipratropium prepared according to the process of any one of clauses 79 to 112.

Clause 117 the amorphous calcium salt of clause 116, which is amorphous obiprifleside calcium.

Clause 118 a method of making amorphous obipratropium calcium salt comprising treating obipratropium with an acid to form a salt, solvate, composition or combination thereof, isolating the salt, solvate, composition or combination thereof, and treating the salt, solvate, composition or combination thereof with a calcium source to make amorphous obipratropium hemi-calcium salt.

Clause 119. The method of clause 118, wherein the calcium source is calcium chloride.

Clause 120. A salt, solvate, composition or combination thereof, comprising obipratropium and a free acid.

Clause 121. The salt of clause 120.

Clause 122. The solvate of clause 120.

Clause 123. The composition of clause 120.

Clause 124 the salt, solvate, composition, or combination thereof of clause 120, wherein the free acid is selected from the group consisting of sulfonic acid, sulfuric acid, halogenated acid, acetic acid, aspartic acid, benzoic acid, bicarbonic acid (bicarbonic acid), acid tartrate, carbonic acid, citric acid, capric acid, fumaric acid, glucoheptonic acid (gluceptic acid), gluconic acid, glutamic acid, glycolic acid, caproic acid, hydroxynaphthoic acid, isethionic acid, lactic acid, lactobionic acid, malic acid, maleic acid, mandelic acid, mucic acid, nitric acid, caprylic acid, oleic acid, pamoic acid, pantothenic acid, phosphoric acid (phosphic acid), polygalacturonic acid, propionic acid, salicylic acid, stearic acid, succinic acid, tartaric acid (TARTRIC ACID), and theachloric acid (teoclic acid), wherein the sulfonic acid may be benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, ethanedisulfonic acid, or methanesulfonic acid, the sulfuric acid is methylsulfuric acid, and the halogenated acid may be HCl, HBr, or HI.

The method of clause 125, 118, wherein the calcium source is a halogenated calcium salt.

Clause 126. The method of clause 118, wherein the calcium source is a soluble calcium salt.

Clause 127. The method of clause 118, wherein the calcium source is a calcium salt.

The present disclosure may be further directed to methods set forth by one or more of the preceding non-limiting clauses.

Examples

Analysis and physical characterization methods:

The methods used throughout the study are summarized in table a. Specific parameters and conditions for analysis and physical assessment of each non-limiting example are set forth in the relevant section of this example.

Table A

XRPD

XRPD analysis was run in transmission mode using standard XRPD eplerate (Aptuit) method on an X 'pert Pro/Empyrean X-ray diffractometer (PANalytical) equipped with an X' celearter detector. The data was evaluated using high-scoring software (Highscore Plus software). The instrument parameters used are listed in table B below.

Table B

Particle Size Distribution (PSD)

PSD analysis was run on a New Pathatch Heroules (Sympatec Helos) laser diffraction instrument equipped with RODOS/M for dispersion and ASPIROS or VIBRI for sample delivery. Powder dispersion is achieved by using compressed air and a spray gun utilizing the venturi effect. Details of the PSD process are set forth in Table C.

Table C

Differential dissolution method at pH 6.8 (Obipcetrap) Table D

Differential dissolution method at pH 4.5 (ezetimibe) Table E

QC dissolution method at pH 6.8 (Obipfrinpi) Table F

Instrument parameters	Value of
		Apparatus and method for controlling the operation of a device	USP apparatus II
Dissolution medium	Phosphate buffer at pH 6.8+0.2% w/v polysorbate 80
		Volume of dissolution Medium [ mL ]	1000
Temperature of dissolution medium [ DEGC ]	37±0.5
		Rotational speed [ rpm ]	75-Infinity point 250rpm
Sampling time [ minutes ]	15, 30, 45, 60 And 70 (70' at 250rpm is the infinity point)
		Sampling volume [ mL ]	1
Separation technique (.)	PVDF 0.45 μm film
		Filter prewet volume [ mL ]	9 (10 ML sample and put the first 9mL back into the container)
Detection of	HPLC-UV

QC dissolution method at pH 4.5 (ezetimibe) Table G

Instrument parameters	Value of
		Apparatus and method for controlling the operation of a device	USP apparatus II
Dissolution medium	Acetate buffer at pH 4.5+0.45% w/v SLS
		Volume of dissolution Medium [ mL ]	500
Temperature of dissolution medium [ DEGC ]	37±0.5
		Rotational speed [ rpm ]	75-Infinity point 250rpm
Sampling time [ minutes ]	15, 30, 45, 60 And 70 (70' is the infinity point).
		Sampling volume [ mL ]	1
Separation technique (.)	ww/PTFE 0.45
		Filter prewet volume [ mL ]	9 (10 ML sample and put the first 9mL back into the container)
Detection of	HPLC-UV

Determination and impurity/related substance (obbesitrapine) table H

Determination and impurity/related substances (ezetimibe) Table I

Example 1

Fixed dose combination tablets of 10mg ezetimibe and 5mg obipratropium (small scale batch approximately 500 g)

High shear granulation and fluid bed drying

Four prototype formulations were evaluated. The excipients contained in the granules were plastic filler (Avicel PH 101), brittle filler (Pharmatose 200M), binder (colestuary 30), disintegrant (glycolys) and surfactant (Kolliphor SLS fine). In the preliminary four tests (particle batches A4459/05/01, A4459/05/02, A4459/05/03 and A4459/05/04), the amounts of plastic and brittle fillers were assessed at high or low levels and two high shear granulation processing conditions were tested. In the last two experiments (particle batches A4459/07/01 and A4459/08/01), the formulations were prepared at high emulsion levels and lower impeller speeds (according to processing condition 2). The composition of the excipients and the method of addition were modified as detailed in table 1.

The material was dispensed at the target weight and ezetimibe, obipratropium and intragranular excipients were manually screened and transferred to a granulation bowl (granulation bowl). The granulation solution is prepared by dissolving the desired excipients in water.

Small-scale pellets were dried using a STREA fluid bed granulator and the material was fluidized in a bowl by adjusting the air volume as needed until the LOD of the dried pellets was equal to or lower than the initial LOD. The intake temperature, product temperature, exhaust temperature, and air flow (air flow volume) were recorded throughout the drying period. After drying, the particles were subjected to particle homogeneity tests of API, LOD, sieve analysis, TBD and XRPD.

Preparation and tabletting of the final blend

The final blend is prepared by precisely weighing the required amount of extra-granular excipient. The excipients (except magnesium stearate (MgSt)) were then manually screened, added to a bin (bin) of appropriate volume along with the granules, and blended using a Pharmatech mixer. MgSt was screened separately and added to the silo. For compression, a single punch (single punch compression machine) (particularly an EK0 tablet press) was used to generate the compression curve and produce tablets with a target weight of 150.0 mg. Based on the information collected for the compression curve, small-scale tablet manufacturing was performed. The tablets were tested by KF and XRPD for appearance, determination and impurity content, differential solubility, ezetimibe USP tablet dissolution method, content uniformity, water content. All the produced intermediates and uncoated tablets were stored in double-layer low density polyethylene (low density polyethylene, LDPE) bags closed with a cable tie and transferred to aluminum bags sealed with silica.

TABLE 1 composition of small-scale 10mg ezetimibe and 5mg of granules and tablets tested with obipratropium (% w/w)

Results:

Development of a small-scale test for tablets of ezetimibe 10mg and obipratropium 5mg

Small scale batches of particle production were successfully performed. During granulation, energy consumption increased after addition of the granulation solution, and after drying, the LOD of the granules was lower than the initial LOD (table 2). Particles A4459/05/01 and A4459/05/02 exhibited coarser particles than particle batches A4459/05/03 and A4459/05/04. This is related to the higher lactose levels in the formulation, and not to the granulation parameters (process condition 1 versus process condition 2). As the amount of binder and water used for granulation decreased and the level of surfactant increased, the particle batches a4459/07/01 and a4459/08/01 (manufactured with higher levels of lactose) exhibited particles containing a greater portion of fines than batches a4459/05/01 and a4459/05/02 (figures 1 and 2). In short, tablets with higher microcrystalline cellulose content (batches A4459/05/07 and A4459/05/08) disintegrate faster, have lower friability and have higher hardness values than tablets with higher lactose amounts (batches A4459/05/05 and A4459/05/06). Overall, these tablets present a suitable appearance in batches. Tablet batches A4459/07/02 and A4459/08/02 (containing high levels of lactose) exhibited faster disintegration times and suitable dissolution profiles for both drug substances. However, the hardness and friability of the tablets cannot be improved to levels considered acceptable due to capping (capping) and failure of the friability test. Tablet hardness was lower than that obtained using the previous test (batches A4459/05/05 and A4459/05/06).

TABLE 2 LOD of 10mg ezetimibe, 5mg small-scale test of obsemitrapib

Particle characterization

Chemical characterization analysis

Homogeneity tests were performed on particles of both obipratropium and ezetimibe, the results are shown in table 3. Both APIs were homogeneously dispersed in the particles, batch a4459/05/03 achieved the maximum RSD% value, but the maximum RSD% value was still within the typical acceptable range of particle homogeneity.

TABLE 3 particle uniformity for 500g batch scale prototype

Characterization of physical Properties

As shown in table 4, small amounts of ezetimibe (EZE) hydrate were found in all wet particle samples. However, during the drying process, the EZE hydrate formed was converted back to anhydrous EZE, except for batch 05/01, which appears to still exist in small amounts of the hydrate polymorphic form.

TABLE 4 XRPD data summary for small-scale 10mg ezetimibe and 5mg obipratropium blends, wet and dry particles

Tablet characterization

Chemical characterization analysis

The characterization results for small-scale batches are shown in table 5. The results of the assay and impurity are expected and the impurity profile is consistent with the API of both inputs. Since the AV values of the content uniformity results are significantly lower than the AV requirements of the pharmacopoeia, it was also found that all prototypes had a homogeneous API content. The water content results were found to be in the range of 4.5% and 5.0%, and no defects were observed in appearance.

The dissolution profile of olanzapine shows similar dissolution trends for prototypes a4459/05/08, a4459/05/06 and a4459/05/07, with prototypes a4459/05/08 (high Avicel content and low impeller speed) dissolving rapidly in the range of 5 to 15 minutes. Prototype a4459/05/05 (high lactose and high impeller speed) dissolved significantly slower. The dissolution results of the USP ezetimibe method at pH 4.5 are consistent with those observed at pH 6.8. For prototype 3 lot a4459/07/02 (4% binder) and prototype 4 lot a4459/08/02 (1% binder), a significant improvement in solubility characterization was observed, indicating that the profile of prototype 4 was consistent with the reference commercial ezetimibe tablet. The dissolution curves are presented in figures 3, 4 and 5. The determination of both obrisetrapib and ezetimibe, content uniformity and impurity profile showed no significant differences between the four formulations.

TABLE 5 analytical characterization results of small-scale 10mg ezetimibe and 5mg obipratropium

Stress stability

Prototype 1 and prototype 2 with different process conditions were evaluated in stress stability studies with the following design

TABLE 6 design of stress stability study

Key:

T = appearance, assay and related substances, differential solubility, water content by KF, and formal check by XRPD

(T) =optional test

The results are reported in tables 7, 8 and 9.

TABLE 7 results of appearance, determination and water content of small-scale 10mg ezetimibe and 5mg obipratropium

TABLE 8 impurity profile of small-scale 10mg ezetimibe and 5mg obipratropium

TABLE 9 results of solubility characterization of stress stability of ezetimibe and 5mg obipratropium at small scale 10mg

TABLE 9 (follow-up) results of solubility characterization of stress stability of ezetimibe at small scale 10mg and olanzapine at 5mg

Characterization of physical Properties

In addition to prototype tablet A4459/05/05, where the presence of small amounts of EZE hydrate was observed in the initial time point samples and all of the stable-placed samples, for the other tablet prototypes, small amounts of EZE hydrate forms were present at the 3 rd week (3 WK) and 4 th week (4 WK) time points. XRPD data are summarized in table 10.

TABLE 10 XRPD data summary for stress stability of small-scale 10mg ezetimibe and 5mg obsemitrapib tablets

Example 2

Fixed dose combination of 10mg ezetimibe and 10mg obipratropium tablet (small scale batch approximately 500 g)

Details of prototype formulations made in this set of experiments are summarized in tables 11 and 12. A key modification of the formulation composition was to increase the dose strength of the obipratropium (free acid) from 5.0mg to 10.0mg.

High shear granulation and fluid bed drying

According to process condition 2, these experiments were performed in small-scale batches (500 g batch size). However, batch A4459/16/02 (referred to as "prototype C scale-up" (prototype C scale-up)) was performed on a batch size scale of 2 Kg.

The powders were manually screened, charged into a granulation bowl and mixed for 5 minutes. The granulation solution is sprayed at the desired spray rate and wet agglomerated prior to drying the material in the fluid bed dryer. The inlet air temperature and air volume are adjusted as necessary to fluidize the dried particles until their LOD is equal to or lower than the initial LOD. The particles were characterized as API content uniformity, LOD (shortly after milling), sieve analysis, TBD and XRPD. Particle lot A4459/13/01 (prototype A) and lot A4459/16/02 (prototype C scale-up) were divided into two aliquotted samples to make the final blend required to produce 150mg tablets and 200mg tablets.

Preparation, tabletting and coating of the final blend

The final blend is prepared by precisely weighing the extra-granular excipients to produce a tablet having the desired composition. The excipients are manually screened and mixed using a silo with the appropriate volume. Lubricants (MgSt) were screened separately and added to the bowl for mixing. A single punch was used to generate the compression curve and to produce small scale batches of tablets. The friability, disintegration time, hardness, appearance and thickness of the tablets were monitored throughout the process. Tablets were tested for differential solubility, ezetimibe USP tablet dissolution method, and XRPD.

Three selected tablet batches (prototype B, prototype C and prototype C200) were coated with 20% w/w Opadry AMB II white aqueous suspension. Coating process parameters and tablet weight gain were monitored throughout the process. The coated tablets were tested for XRPD, differential solubility of obrisetrapib, and ezetimibe USP tablet dissolution methods at a paddle speed of 75 rpm.

All the intermediates and final drug products produced were stored in double-layer LDPE bags closed with ties and transferred to heat-seal aluminum bags containing silica.

TABLE 11 composition of granules and tablets of small-scale 10mg ezetimibe and 10mg obipratropium (% w/w)

* No water present in the tablet, inner = material added as dry powder, outer = material dissolved in water for granulation;:. Due to equipment problems, the addition is done in lower amounts

Table 12 composition of granules and tablets of 10mg ezetimibe and 10mg obipratropium (% w/w)

* Water is not present in the tablet; inner = material added as dry powder, outer = material dissolved in water for granulation results:

These granulation tests were successfully performed. Overall, the PSD of the particles was similar to that of batch a4459/08/01 (prototype 4) and showed a relatively large number of fine particles (fig. 18). The tablet lot disintegrated under similar compression forces for a comparable time, with higher hardness and lower friability value than lot a4459/08/02 (prototype 4, condition 2). Tablets do not exhibit any serious drawbacks (e.g., capping, delamination). Upon careful examination, the coated tablets exhibited a smooth white surface without any visual appearance defects.

Particle characterization

Chemical characterization analysis

Homogeneity tests were performed on particles of both obipratropium and ezetimibe and the results are shown in table 13. Analysis was performed at n=6, except that the amplification lot was performed at n=10.

Characterization of physical Properties

XRPD data for developing prototypes are summarized in table 14. Prior to or during the pelletization process, EZE hydrate may be observed in the sample. However, the amount of EZE hydrate detected appears to be always very limited.

Tablet characterization

Chemical characterization analysis

The characterization results for small-scale batches are shown in table 15. Dissolution testing was performed on the prototype. The dissolution results of olanzapine show that all prototypes tested have similar profiles with minor differences that are considered as analytical variability. Prototype D and prototype C gave the most promising results for ezetimibe, with prototype C achieving the promising results meeting USP specification q=80+5 in 30 minutes in three containers. This most promising prototype was also characterized using the USP dissolution method conditions of ezetimibe at higher paddle speeds of 75 rpm. This is because the USP method developed for lighter tablets appears to oversubscribe tablets of up to 200mg in weight target as a result of emphasis compared to the fixed dose combinations already developed. The results show a profile consistent with that of the currently commercialized formulation.

Characterization of physical Properties

All of the small scale prototype tablets produced contained small amounts of EZE hydrate except prototype C and C shown in table 16, which were scaled up to 200mg batches.

TABLE 13 particle uniformity of 10mg ezetimibe and 10mg (free acid) obipratropium development prototype

TABLE 14 XRPD data summary for 10mg ezetimibe and 10mg (free acid) obipratropium development prototype

TABLE 15 results of solubility characterization of small-scale 10mg ezetimibe with 10mg obipratropium (free acid)

TABLE 16 summary of XRPD data for prototype batches of small-scale tablets

Sample of	Batch ID	XRPD
			Tablet prototype A	A4459/14/01	OBI+Anhydrous EZE+ small amounts of EZE hydrate
Tablet prototype A200 mg	A4459/14/04	OBI+Anhydrous EZE+ small amounts of EZE hydrate
			Tablet prototype B	A4459/14/02	OBI+Anhydrous EZE+ small amounts of EZE hydrate
Tablet prototype C	A4459/14/03	OBI+Anhydrous EZE
			Tablet prototype C magnification	A4459/16/03	OBI+Anhydrous EZE
Tablet prototype C zoom in 200mg	A4459/18/03	OBI+Anhydrous EZE+ small amounts of EZE hydrate
			Tablet prototype C2% SLS	A4459/18/01	OBI+Anhydrous EZE+ small amounts of EZE hydrate
Tablet prototype D	A4459/18/02	OBI+Anhydrous EZE+ small amounts of EZE hydrate

Stress stability

Based on the process tabletting parameters and solubility data, the following tablet prototypes were selected to assess the feasibility of the coating process:

a4459/16/03 (150 mg/tablet, prototype C amplified)

A4459/18/03 (200 mg/tablet prototype C magnification/200 mg),

And then stress stability studies were performed with the following design for stability settings. The results are shown in tables 17 and 18.

TABLE 17 design of stress stability study

Key:

t = differential solubility of appearance, assay and related substances, content uniformity (only at initial stage), water content by KF, and formal check by XRPD

(T) =optional test

TABLE 18 results of small-scale measurement of stress stability, water content and visual appearance for ezetimibe at 10mg and olanzapine (free acid) prototype C200 and prototype C at 10mg

TABLE 19 impurity profile results for small-scale 10mg ezetimibe with 10mg obsemitrapib (free acid) prototype C200 and prototype C amplified stress stability

N.d. =undetected

TABLE 20 solubility characterization results of small-scale 10mg ezetimibe with 10mg obsemitrapib (free acid) prototype C200 and prototype C amplified stress stability

Characterization of physical Properties

XRPD data for samples with stress stability when placed are summarized in table 21. Both prototype tablets exhibited ezetimibe water compound when exposed to 40 ℃ per 75% rh for 2 weeks, however, once packaged, no polycrystallme transformation occurred until 4 weeks of storage.

Table 21 prototype C200 with stress stability when placed and amplified XRPD data summary

Example 3

Fixed dose combination of 10mg ezetimibe and 10mg obipratropium obtained by co-granulation of a pharmaceutical substance/active ingredient (FDC 1) (small-scale batch about 500 g)

High shear granulation, drying, preparation of the final blend and tabletting

Three compositions (composition 1 lot A4459/20/02, composition 2 lot A4459/20/03, and composition 3 lot A4459/20/04) were prepared as summarized in Table 22. The prototype composition selected for these compositions was the prototype composition of "prototype C" (e.g., particle lot A4459/13/03). The preparation and characterization of the particles (LOD and XRPD) is set forth in the previous section (small scale manufacturing). The particles were tested for content uniformity, LOD, sieve analysis, TBD and XRPD. As described in the previous example, the manufacture of blends for tabletting and compression curves and small batches of tablets with a tablet weight of 150mg was performed. Tablets were tested for content uniformity, XRPD, solubility and water content by KF. All intermediates and final pharmaceutical products produced were stored as described in the previous section.

TABLE 22 composition of granules and tablets of FDC1 composition (% w/w)

* Water is not present in the final product. All excipients used for granulation are added in dry powder form

Results

High shear granulation was successfully performed. The drying step was performed without any problem and after 15 minutes of drying the pellet had a LOD lower than the initial LOD. In general, the granules exhibited a relatively large number of fine particles despite an increase in impeller speed (composition 1) (fig. 26), wet agglomeration time (composition 2), or amount of granulating agent (composition 3). The tablet friability, disintegration time, thickness and hardness of these tablet batches were found to be similar.

Characterization of particle chemistry

The particles were subjected to homogeneity tests and it was found that obrisetrapib and ezetimibe were homogeneously dispersed.

Characterization of physical Properties

XRPD data for the blend/particle prototype FDC1 process are summarized in table 23. Eze hydrate was only present in the wet particulate sample. All three prototypes showed similar flowability.

Tablet chemical characterization

The results of the chemical characterization of the FDC1 tablets are shown in table 24. The results of the analytical characterization did not show any significant differences between the three compositions.

Characterization of physical Properties

XRPD data from tablets of FDC1 composition are summarized in table 25. Eze hydrate was absent in all samples.

TABLE 23 summary of XRPD data for particles from FDC1 compositions

TABLE 24 chemical characterization results of FDC1 compositions

Table 25 summary of XRPD data for particles of FDC1 composition

Example 4

Fixed dose combination (FDC 2) of 10mg ezetimibe and 10mg obipratropium obtained by granulating ezetimibe and adding obipratropium outside the granulation

High shear granulation and drying

Three compositions were prepared as summarized in table 26. The excipients contained in the granules are the same as those used in the manufacture of granules for the FDC1 process. The formulation composition of these particles reflects the formulation composition of the FDC1 particle "prototype C". The methods of high shear granulation, granule drying and milling have been described in the previous section. The content uniformity (ezetimibe only), sieve analysis, TBD and XRPD of the particles were tested.

Preparation, tabletting and coating of the final blend

The composition of the external preparation for granules is as follows:

-obixitracin

Plastic filler (Avicel PH 200)

Brittle filler (Pearlitol 200 SD)

-Disintegrants (Glycolys)

Glossia (Glydant) (Aerosil 200)

-Lubricant (LIGAMED MF-2-V)

The final blend was prepared by precisely weighing the extra-granular components (excipients and API) and screening them. The excipients and granules were loaded into bins of appropriate volume and blended using a Pharmatech mixer. Then, a lubricant (MgSt) was added to the silo and mixed.

To generate the compression curve and to produce small batches of tablets, a single punch (EK 0) equipped with a 9.0mm round punch (r=11) was used. The target tablet weight was 230mg and throughout the process, tablet friability, disintegration time, hardness, appearance and thickness, as well as individual tablet weights and ten tablet weights were monitored.

Tablets were tested for content uniformity (layering samples: beginning of production, middle and end), XRPD, solubility, and water content by KF.

Tablets were coated with a 20% w/w Opadry AMB II white aqueous suspension at the desired target weight increase (target weight increase 3% w/w, limit of 2% w/w to 4% w/w). Coating suspensions and coating methods are described in the previous sections. Coating parameters and tablet weight gain were monitored throughout the process. The coated tablets were tested for XRPD, solubility, appearance, content uniformity, and water content by KF.

All the intermediates and final drug products produced were stored in double layer LDPE bags with silica and transferred to heat sealed aluminium bags.

TABLE 26 composition of granules, tablets and coated tablets of FDC2 composition (% w/w)

Results

High shear granulation of the FDC2 composition was successfully performed. The drying step was performed without any problem and after about 16 minutes of drying the pellet had a LOD lower than the initial LOD. The particles showed a relatively large number of fine particles (fig. 29). The values of disintegration time and thickness between FDC2 tablet batches are similar.

Characterization of particle chemistry

The homogeneity of both obrisetrapib and ezetimibe of the a4459/20/01 blend was tested and found to be homogeneously dispersed. The results of the other two particles were not collected.

Tablet chemical characterization

The chemical characterization results of prototype 1 of the FDC2 tablets are shown in tables 27 and 28. The results of the analytical characterization did not show any significant differences between the three prototypes of FDC 2.

Characterization of physical Properties

XRPD data from blends/particles of FDC2 compositions are summarized in table 29. XRPD data for tablets from the FDC2 process are summarized in table 30. A small amount of Eze hydrate was present in prototype 1.

TABLE 27 analytical characterization of uncoated FDC2

TABLE 28 analytical characterization of coated FDC2

TABLE 29 summary of XRPD data for particles from FDC2 compositions

Sample of	Batch ID	XRPD
			Prototype 1 final dry particles	A4459/20/01	OBI+Anhydrous EZE+ small amounts of EZE hydrate
Prototype 2 final dry particles	A4459/25/01	OBI+Anhydrous EZE
			Prototype 3 final dry particles	A4459/25/02	OBI+Anhydrous EZE

TABLE 30 summary of XRPD data for tablets from FDC2 compositions

Stress stability

The stress stability study was performed on the prototype 2 coated tablets using the following design

TABLE 31 design for stress stability study

Key:

(T) =optional test

The results are recorded in tables 66, 67 and 68.

TABLE 32 determination of stress stability, water content and visual appearance of prototype 2FDC2 results

TABLE 33 impurity profile results for prototype 2FDC2 stress stability

N.d. =undetected

TABLE 34 dissolution characterization results of prototype 2FDC2 stress stability

Example 5

Fixed dose combination (FDC 3) of 10mg ezetimibe and 10mg obipratropium obtained by granulating obipratropium and adding ezetimibe outside the granulate

Prototype compositions were prepared as summarized in table 35. The granulation process was as described above for process condition 2. The methods of high shear granulation, granule drying and milling have been described in the previous section. The content uniformity (obbestrepide alone), sieve analysis, TBD and XRPD of the particles were tested.

Preparation, tabletting and coating of the final blend

To generate the compression curve and to make small batches of tablets, a single punch is used. The target tablet weight was 230mg and throughout the process, tablet friability, disintegration time, hardness, appearance and thickness, as well as individual tablet weights and ten tablet weights were monitored. Tablets were tested for content uniformity (layering samples: beginning of production, middle and end), XRPD, solubility, and water content by KF.

Tablets were coated with a 20% w/w Opadry AMB II white aqueous suspension at the desired target weight gain. Coating suspensions and coating methods are described in the previous sections. Coating parameters and tablet weight gain were monitored throughout the process. The coated tablets were tested for XRPD, solubility, appearance, content uniformity, and water content by KF.

TABLE 35 composition of granules, tablets and coated tablets of FDC3 composition (% w/w)

* Lactose or lactose alternative example 6

Fixed dose combination (FDC 4) of 10mg ezetimibe and 10mg of obipratropium as bilayer tablets obtained by granulating obipratropium and ezetimibe, respectively, followed by compression

Prototype compositions were prepared as summarized in table 36. The granulation process of ezetimibe is the same as described above for FDC1 and the granulation process of obrisetrapib is the same as described above for FDC 3. The method of high shear granulation, granule drying and milling is the same as described in the previous section for FDC 1. The granules were then fed into the compressor via two hoppers (hopper). The mold is filled with the first particles and then slightly compressed. The second particles are then filled and subsequently compressed as explained in the previous examples. Individual particles were tested for content uniformity (obrisetrapib or ezetimibe), sieve analysis, TBD and XRPD. The granules were compressed to form tablets according to the method described above. Tablets were tested for content uniformity (layering samples: beginning of production, middle and end), XRPD, solubility, and water content by KF.

TABLE 36 composition of granules, tablets and coated tablets of FDC4 composition (% w/w)

* Purified water is a granulating agent and does not play an important role in any final formulation example 7

FDC1 composition and amplification of FDC2 composition

High shear granulation, drying, final blend, tabletting and coating

The API and excipients were accurately partitioned, screened and added to the granulation bowl according to the methods detailed above in the FDC1 and FDC2 formulation methods. The granulation parameters were the same for both the FDC1 and FDC2 processes. The content uniformity (for FDC1 method only), LOD, sieve analysis, TBD and XRPD of the particles were tested.

Final blends of FDC1 and FDC2 compositions were prepared to make tablets with lot numbers and compositions detailed in table 37. The extra-granular components are manually screened and loaded into bins of appropriate volume. The particles were mixed with the extra-granular material using a Pharmatech mixer. Then, the content uniformity of the FDC2 blend was tested.

The use of rotary presses (rotation PRESS MACHINE) was assessed for the purpose of generating compression curves and performing tabletting exercises. Tablet friability, disintegration time, hardness, thickness and appearance, individual tablet weights and ten tablet weights were monitored during tabletting exercises. Tablets were tested for content uniformity (layering samples: beginning of production, middle and end), XRPD, solubility, and water content by KF.

Tablets were coated with a 20% w/w Opadry AMB II white aqueous suspension at the desired target weight increase (target weight increase 3% w/w, limit of 2% w/w to 4% w/w). Coating suspensions and coating methods are described in the previous sections. Coating parameters and tablet weight gain were monitored throughout the process. The coated tablets were tested for XRPD, solubility, appearance, assay, and impurities, and for water content by KF.

TABLE 37 composition of granules, tablets and coated tablets for FDC1 and FDC2 magnification test (% w/w)

* Water is not present in the final product. Excipients for granulation are added in the form of dry powders

Results

High shear granulation of the enlarged batch was successfully performed. By sieving, the particles exhibited similar PSD values and contained a large amount of fines (fig. 37). These PSD values are comparable to those found in previous experiments (e.g., batch A4459/16/02 as a reference for FDC1 amplified batch and batch A4459/25/01 as a reference for FDC2 amplified batch). Despite this similarity in PSD data, the flowability of the amplified lot is improved over that of the reference lot according to the value of the millipore ratio (Hauser ratio). These tablets have a disintegration time of less than 5 minutes and a friability of less than 0.2%. The coating process was performed without any serious problems, and the appearance of the coated tablets of both formulation methods was suitable because the surface of the tablets was smooth.

Characterization of particle chemistry

Both the particles and the final blend were tested for homogeneity of both obrisetrapib and ezetimibe and found to be homogeneously dispersed. The solubility results of both obrisetrapib and ezetimibe and the impurity profile results of ezetimibe are shown in table 38, fig. 39, and fig. 40.

Tablet chemical characterization

The results of the chemical characterization of the enlarged batch of uncoated tablets are shown in table 39. The dissolution results for uncoated tablets under different compression forces are shown in tables 40, 41, 42 and 43. The results of coating the tablets are shown in table 42.

Characterization of physical Properties

XRPD data from the magnified batches of particles/tablets are summarized in table 43. All batches tested showed no presence of EZE hydrate.

TABLE 38 amplified batch particle and final blend impurity profile and dissolution results

TABLE 39 analytical characterization results for uncoated amplified lots

TABLE 40 Ezetimibe dissolution results for amplified batch A4459/29/02 (FDC 1) under different compression forces

TABLE 41 amplified results of dissolution of Orbicubic from batch A4459/29/02 (FDC 1) under different compression forces

TABLE 42 Ezetimibe dissolution results for amplified batch A4459/29/05 (FDC 2) under different compression forces

TABLE 43 amplified dissolution results of Orbicubic from batch A4459/29/05 (FDC 2) under different compression forces

TABLE 44 analytical characterization results for coating enlarged batch 10mg ezetimibe 10mg obbestrepide

TABLE 45 summary of XRPD data for particles/tablets from a magnified batch

Example 8

Technical batches of FDC1 and FDC2 compositions

The lot numbers and compositions of granules, tablets and coated tablets produced by technical manufacturers of FDC1 and FDC2 formulations are detailed in table 49.

High shear granulation and drying

The components of FDC1 pellets (batch A4459/30/02) and FDC2 pellets (batch A4459/30/01) were manually screened and added to the pelletization bowl. For both the FDC1 and FDC2 compositions, the batch size of the granules was 2kg and the same granulation processing conditions were used. The physical mixture was blended at 220rpm for 5 minutes and the LOD test was performed. The granulating agent (purified water) was sprayed and after spraying, wet agglomeration was performed for 1 minute. Samples for XRPD and LOD were taken. The granules were then dried using a fluid bed dryer. The drying step is ended when the LOD of the pellet is equal to or lower than the initial LOD or lower than 3% (w/w). The content uniformity (for FDC1 only), PSD, LOD, sieve analysis, TBD and XRPD of the particles were tested.

Final blend and tableting

The method of preparing the final blend is set forth in the previous section (enlarged batch). The content uniformity of the FDC2 blend was tested.

To generate the compression curve and perform the tabletting activities, a rotary press was used. For the manufacture of FDC1 tablets (150 mg target tablet weight), a 7.0mm diameter punch was used, while for the manufacture of FDC2 tablets (230.0 mg tablet weight), an 8.5mm diameter punch was used. Tablet friability, disintegration time, hardness, thickness and appearance, individual tablet weight and ten tablet weight were monitored during tabletting. Tablets were tested for content uniformity (layering samples: beginning of production, middle and end), XRPD, solubility, and water content by KF.

Coating layer

A coating suspension with a solids content of 20% (w/w) was prepared by adding the required amount of Opadry to water while stirring. The suspension was mixed for not less than 45 minutes and its visual homogeneity was confirmed. The spray rate of the coating suspension was then measured. The coating suspension was kept stirring at all times. The weight increase of the tablets was monitored throughout the manufacture and spraying was stopped when the desired target increase (3% w/w,2% w/w to 4% w/w limit) was achieved. The coated tablets were visually inspected and tested for XRPD, solubility, appearance, content uniformity, and water content by KF.

All the intermediates and final pharmaceutical products produced were stored in a stabilization chamber with the following packaging:

● 60mL of high density Polyethylene (HIGH DENSITY Polyethylene, HDPE) was induction sealed and closed by a child safety cap. The filling count of the tablets was 20;

● A2 g desiccant canister was used for 60mL High Density Polyethylene (HDPE) induction sealing and closed by a child safety cap. The filling count of the tablets was 20.

TABLE 49 composition of granules, tablets and coated tablets of FDC1 and FDC2 technical batches (% w/w)

Results

Technical batches of FDC1 and FDC2 compositions

High shear granulation of technical batches was successfully performed and no problems occurred during the process. The powder consumption was comparable to that observed in the scaled-up batch. Since the LOD of the pellets was below 3%, the drying process was performed in 45 minutes. The particles showed similar PSD values and the amount of fine particles was relatively large (e.g., the amount of particles less than 125 μm was about 70% to 73%) (fig. 45). No relevant differences were observed compared to the scaled-up batch.

Compared to the FDC2 scaled-up batch (batch a 4459/29/06), the FDC2 technical batch provided harder tablets despite similar compression force levels, with very similar disintegration times between these batches for a given tablet hardness value.

Characterization of particle chemistry

Both the particles and the final blend were tested for homogeneity of both obrisetrapib and ezetimibe and found to be homogeneously dispersed.

Tablet chemical characterization

Chemical characterization results for technical batch coated tablets are shown in table 50. Tablets exhibit suitable quality levels that claim that% are within typical acceptance criteria for the clinical stage (i.e., 90.0% to 110.0%). The content uniformity test results meet the pharmacopoeia requirement AV <15.0. At 45 minutes, the solubility met the proposed specification of q=75% (fig. 46 and 47).

Characterization of physical Properties

XRPD data for particles/tablets from technical batches are summarized in table 51. All batches tested, except the particles from the FDC2 formulation, showed no presence of EZE hydrate. However, the hydrate form disappeared in the coated tablet. The two batches of particles showed similar flowability. The PSD data for the particles are shown in Table 52 and FIG. 48. The batch shows a similar bimodal curve (bi-modal curve).

Stability study

An overview of the physicochemical analysis of the technical batch after three months of storage of the FDC2 composition is provided in table 53 (without desiccant) and table 54 (with desiccant), and an overview of the physicochemical analysis of the technical batch after three months of storage of the FDC1 composition is provided in table 55 (without desiccant) and table 56 (with desiccant).

TABLE 50 analytical characterization results for coating technical batches

TABLE 51 summary of XRPD data for particles/tablets from technical lots

TABLE 52 summary of PSD data for particles from technical lots

TABLE 53 Long term and accelerated storage stability results for FDC2 technical lot A4459/31/01 (without desiccant)

Watch 53 (Xuezhi)

N/A-inapplicability

TABLE 54 Long term and accelerated storage stability results for FDC1 technical lot A4459/31/02 (without desiccant)

Watch 54 (Xuezhi)

N/A-inapplicability

TABLE 55 long-term and accelerated storage Condition results for FDC2 technical lot A4459/31/01 (desiccant contained)

Watch 55 (Xuezhi)

TABLE 56 long-term and accelerated storage Condition results for FDC1 technical lot A4459/31/02 (desiccant contained)

Watch 56 (Xuezhi)

N/A-inapplicability

Example 9

A study was conducted to evaluate the comparative bioavailability of olanexidine/ezetimibe 10mg/10mg (FDC 1 and FDC 2) and two fixed dose combination formulations co-administered with olanexidine 10mg and ezetimibe 10mg in healthy adult subjects under fasted conditions

Study design

This is an open, single dose, randomized, three treatment, three cycle, six sequence crossover study that compares co-administration of two test and reference products under fasted conditions. At each study period, subjects received either treatment T1 (1X obbestrapib 10mg and ezetimibe 10mg FDC1 tablet [ formulation #1 ]), treatment T2 (1X obbestrapib 10mg and ezetimibe 10mg FDC2 tablet [ formulation #2 ]), or treatment R (1X obbestrapib tablet 10mg and 1X) after an overnight fast of at least 10 hours(Ezetimibe) tablet 10mg co-administration). The order of administration followed a six-sequence randomization schedule. Blood samples were collected at time intervals exceeding 336 hours before and after dosing during each study period. In each study period, subjects were limited to clinical facilities starting at least 10 hours prior to dosing until 24 hours after dosing, and returned to clinical facilities for blood sample collection at 48 hours, 72 hours, 96 hours, 144 hours, 192 hours, 240 hours, and 336 hours after dosing. The interval between doses was at least 49 days.

The plasma concentrations of obrisetrapib, ezetimibe and its metabolite ezetimibe glucuronide were measured by a fully effective analytical method. Statistical analysis was performed using the mean bioequivalence (bioequivalence) method to evaluate the bioavailability of co-administration of each test formulation relative to the reference product.

Selection of study population

The test population included 36 adult male and female subjects who were healthy, who did not use tobacco, and who did not use nicotine.

Therapeutic administration

Subjects received treatment T1, treatment T2, or treatment R and were directly observed following an overnight fast of at least 10 hours according to a randomized schedule of three treatments, three cycles, six sequences (table 57).

● Treatment T1:1xObipratropium 10mg and ezetimibe 10mg FDC tablet (formulation # 1)

● Treatment T2:1xObipratropium 10mg and ezetimibe 10mg FDC tablet (formulation # 2)

● Treatment of 1X obiprift tablet 10mg and 1X(Ezetimibe) tablet 10mg co-administered

Table 57

Sequence(s)	Cycle 1	Cycle 2	Cycle 3
				1	T1	T2	R
2	T2	T1	R
				3	R	T1	T2
4	T1	T2	R
				5	T2	T1	R
6	R	T1	T2

Each dose was administered using 240mL of room temperature water. The subject was instructed to swallow the entire tablet without chewing or biting.

Sample collection, handling and biological analysis planning

Sample size

4ML of collection (K2 EDTA vaccum blood collection tube) for analysis of olanzapine

4ML of collection (K2 EDTA Vaccum blood collection tube) for analysis of ezetimibe and ezetimibe glucuronide

Collection time

Samples prior to quantitative administration were collected within 60 minutes prior to quantitative administration. All times were relative to the dosing minutes.

Analysis for olanzapine before dosing (0 hours) and at 0.50 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, 4.0 hours, 4.5 hours, 5.0 hours, 6.0 hours, 7.0 hours, 9.0 hours, 12.0 hours, 16.0 hours, 20.0 hours, 24.0 hours, 48.0 hours, 72.0 hours, 96.0 hours, 144.0 hours, 192.0 hours, 240.0 hours and 336.0 hours after dosing (% return sample)

Analysis of ezetimibe and ezetimibe glucuronide before (0 hour) and at 0.25 hours, 0.50 hours, 0.75 hours, 1.0 hours, 1.333 hours, 1.667 hours, 2.0 hours, 2.5 hours, 3.0 hours, 3.5 hours, 4.0 hours, 4.5 hours, 5.0 hours, 6.0 hours, 7.0 hours, 9.0 hours, 12.0 hours, 16.0 hours, 20.0 hours, 24.0 hours, 48.0 hours, 72.0 hours and 96.0 hours after the dosing (/ return to the sample)

Total number of collections per cycle/subject 49

Total blood volume per subject total volume collected for pharmacokinetics was approximately 588mL.

Sample processing for analysis of obrisetrapib blood samples were collected in 4ml k2edta evacuated blood collection tubes at room temperature. After collection, the samples were mixed by gently inverting the tube several times (i.e., 8 to 10 times) and placed in an ice/water bath. The sample was then placed in a centrifuge and spun at 3000rpm for 10 minutes at 4 ℃. The resulting plasma was split into two aliquots (at least 1.0mL was present in aliquot 1 and the remainder was present in aliquot 2), transferred to polypropylene sample storage tubes, and stored at-70 ℃ (±10 ℃) until ready for transport to the bioanalytical laboratory. An aliquot of plasma was placed in the refrigerator within 30 minutes after sample collection. After collection, the blood/plasma samples were kept cool in an ice/water bath until placed in a refrigerator.

Sample treatment for analysis of ezetimibe and ezetimibe glucuronide blood samples were collected in 4ml k2edta evacuated blood collection tubes at room temperature. After collection, the samples were gently mixed by inverting the tube several times (at least 8 times) and placed in an ice/water bath. The sample was then placed in a centrifuge and spun at 3000rpm for 10 minutes at 4 ℃. The resulting plasma is split into two aliquots (at least 1.0mL is present in aliquot 1 and the remainder is present in aliquot 2) and transferred to a polypropylene sample storage tube (e.g., sarstedt) # 60.546) and stored at-70 ℃ (or lower) until ready for transport to the bioanalytical laboratory. After collection, the blood/plasma samples were kept cool in an ice/water bath until placed in a refrigerator.

Pharmacokinetic analysis

For all treatments, the following pharmacokinetic parameters of obrisetrapib, ezetimibe and its metabolite ezetimibe glucuronide were calculated.

Primary PK parameters

Cmax, maximum measured plasma concentration.

AUC _0-t the area under the plasma concentration versus time curve calculated by the linear trapezoidal method from time 0 to the last measurable plasma concentration.

AUC _0-∞ the area under the plasma concentration versus time curve from time 0 to infinity, where auc0- +=auc0-t+ct/λz. Ct is the last measurable concentration and λz is the end-point elimination rate constant.

Helper PK parameters

Tmax, time to maximum measured plasma concentration. If the maximum occurs at more than one point in time, tmax is defined as the first point in time having this value.

Λz apparent first order terminal elimination rate constant (APPARENT FIRST-order terminal disposition rate constant). This parameter is obtained by most preferably fitting a least squares linear regression analysis to the terminal linear concentration-time data (TERMINAL IN-linear concentration-time data), calculated from the negative values of the data set slope. The number of data points (3 or more) at the end phase (excluding Cmax) is included in the final regression analysis for determining an evaluable λz from a data set with a highest post-adjustment R square (Rsquared) (R ²) value of 0.7 or greater. λz is considered to be non-evaluable if (1) the last three end points are used to determine λz, and the middle or last point is higher than the previous point, or (2) the resulting adjusted R ² value is less than 0.7.

An evaluable λz is considered unreliable and unreported if the resulting apparent first order terminal half-life (t 1/2) value is longer than the time interval between approximately λz. If the resulting t1/2 value is longer than the time interval during which λz is determined, then a longer interval than the approximated t1/2 is explored. The interval with the next highest adjusted R ² value is selected and the decrease in the adjusted R ² value is assessed to determine if reliable approximation of λz can be made. If λz is deemed unreliable, then the t1/2 and AUC0- ≡values for the dataset are not reported.

T1/2 Primary terminal elimination half-life was calculated as ln (2)/λz

Data set for analysis and statistical methods

The actual time of sample collection was used to provide a linear and semilogarithmic plot of the concentration versus time profile for each subject. Actual sample collection times were used to calculate pharmacokinetic parameters. Plasma concentration data from all evaluable subjects without significant protocol bias were used to approximate Cmax and/or AUC for at least two study periods, one of which included treatment R.

PK parameters from any subjects experiencing emesis within twice the median Tmax of olanzapine or ezetimibe, respectively, calculated from the observations of the particular treatment group (TREATMENT ARM) were excluded from the statistical analysis of the corresponding analytes.

Analysis of variance was performed on AUC0-t, AUC0- ≡and Cmax of post-conversion (In-converted) using an analysis of variance model (analysis of variance model, ANOVA). ANOVA of treatment T1 versus treatment R analysis and treatment T2 versus treatment R analysis were performed using an incomplete block design (incomplete block design), respectively. Treatment T2 was excluded from ANOVA to compare treatment T1 to treatment R, and treatment T1 was excluded from ANOVA to compare treatment T2 to treatment R.

Confidence intervals (90%) were constructed for comparing the geometric mean ratio (obtained from log-transformed data) of AUC0-T, AUC0- ≡and Cmax for each FDC formulation (Tl and T2) to treatment R to test two single-sided hypotheses at a significance level of a=0.05.

Results

Confidence intervals (90%) of geometric mean ratios of AUC0-T, AUC0- ≡and Cmax of olanzapine, ezetimibe and ezetimibe glucuronide (T1 and T2 from formulation #1 and formulation # 2) were found to be in the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of AUC0-T, AUC0- ≡and Cmax of olanzapine, ezetimibe and ezetimibe glucuronide, respectively. Test formulations #1 and #2 were found to be bioequivalent to the reference treatment group (R). Adverse events observed in either the T1 or T2 treatment groups were statistically not significantly different from those observed in the R groups.

Example 10

Phase 2B clinical trial (ROSE 2; NCT 05266586)

1 Introduction and background information

Dyslipidemia is a disease of lipoprotein metabolism, including overproduction or deficiency of lipoproteins, which can be manifested by elevated serum total cholesterol, low Density Lipoprotein (LDL) cholesterol (LDL-C) and Triglyceride (TG) concentration levels, and reduced High Density Lipoprotein (HDL) cholesterol (HDL-C) concentration levels. These diseases are usually diagnosed by measuring serum lipids and classified by an up/down profile of lipid/lipoprotein fractions. Dyslipidemia itself generally does not cause any symptoms, but it can lead to symptomatic vascular diseases, including coronary artery disease and peripheral artery disease. It is well known that although there are many genetic and lifestyle factors leading to the development of vascular disease, dyslipidemia is one of the most prominent risk factors, and that normalization of the lipid profile has been the primary goal of Cardiovascular (CV) protection strategies.

Statin drugs are often the first drugs to treat dyslipidemia. Statin drugs are considered the most potent, most potent and most tolerogenic drugs for lowering LDL-C levels. Many patients, despite treatment with high intensity statin therapies, do not achieve acceptable LDL-C levels with statin alone.

There is a need for a chronic therapy that significantly reduces elevated LDL-C levels when used as an adjunct to high intensity statin therapy.

1.1 Inhibitors of cholesterol ester transfer protein

Cholesterol Ester Transfer Protein (CETP) is a plasma glycoprotein produced in liver and adipose tissue. It circulates in the blood, binds mainly to HDL-C, and is involved in transfer of cholesterol esters and TG between lipoproteins. In particular, it mediates the transfer of cholesterol esters from HDL to particles containing apolipoprotein B (ApoB) (e.g., very low density lipoprotein and LDL-C) to exchange TG. Thus, cholesterol esters from HDL are taken up by the liver via scavenger receptor class B type 1, which also causes HDL-C to be reduced and eventually LDL-C to be raised.

Inhibition of CETP activity decreases ApoB and LDL-C and increases HDL-C. CETP inhibition therapies were originally developed based on the premise that elevated HDL-C levels could prevent CV events. However, clinical studies and Mendelian randomization data (MENDELIAN RANDOMIZATION DATA) have revealed that these effects are caused by changes in the concentration of ApoB-containing particles (including LDL particles) rather than changes in HDL-C levels. Thus, LDL-C and ApoB lowering effects (arising from CETP inhibition and occurring through upregulation of LDL receptors) would be beneficial to patients with elevated LDL-C and increased CV risk.

Ference and colleagues have recently studied the link between changes in LDL-C levels (and other lipoproteins) and the risk of CV events associated with CETP gene variation alone and with a combination of 3-hydroxy-3-methylglutaryl-coa reductase (HMGCR) gene variation.

The results of these mendelian randomization analyses indicate that treatment with CETP inhibitors may be effective in reducing the risk of CV events. Both genetic and therapeutic inhibition of CETP resulted in consistent changes in LDL-C levels versus ApoB levels in number. Further Mendelian randomization analysis led to the conclusion that the difference per unit was lower

The clinical benefit of LDL-C levels may be particularly relevant to the absolute reduction of ApoB-containing lipoprotein particles.

1.2 Obipsilate (TA-8995)

Orbicifacipimox (TA-8995) is a selective CETP inhibitor. The inhibition of CETP by obiprapib blocks the transfer of cholesterol esters from non-atherogenic HDL particles to particles in the atherosclerosis-causing lipoprotein fraction (including LDL), and reduces the concentration of cholesterol in LDL and other atherogenic lipoproteins. Olanzapine also has several additional compound-specific activities that are presumed to be beneficial to the patient. In a recent study, the treatment with obipfrip not only reduced the number of ApoB-containing particles constituting LDL-C, but also increased apolipoprotein E (ApoE), thereby removing cholesterol through the liver and reducing lipoprotein (a) (Lp [ a ]). 5 finally, obipratropium not only effectively increases the concentration of HDL-C and apolipoprotein A1 (ApoA 1) containing lipoproteins, but also proved to be a potent inducer of cholesterol efflux, which is the primary driver of cholesterol antiport. Such an effect is considered important because it is expected to reduce the burden of the formed atheroma.

1.3 Clinical development of obipratropium

Two studies were performed in healthy volunteers, a single escalation dose (TA-8995-01) study and a multiple escalation dose (TA-8995-02) study. A formal and comprehensive QT/QTc study (TA-8995-04) showed that olanzapine had no effect on QTcF. One drug-drug interaction study (TA-8995-05) showed that obipratropium had no significant effect on P-glycoprotein activity, but that obipratropium was a mild inducer of cytochrome P4503 A4. A mass balance study in healthy men led to the conclusion that olanzapine was stably absorbed and the main excretion route was in faeces (TA-8995-07). Finally, the bioequivalence between the obiprift capsule and the tablet formulation was studied (TA-8995-08).

The first patient study conducted was a phase 2 clinical study (TA-8995-03) conducted in denmark and the netherlands, wherein the objective was to evaluate the most preferred doses of obrisetrapib alone and in combination with statins in patients with mild dyslipidemia. The conclusion from this study was that the 10mg daily dose of olanzapine therapy resulted in 45.4% decrease in LDL-C, 179.0% increase in HDL-C, 63.4% increase in ApoA1 and a significant increase in HDL-C efflux capacity.

Furthermore, the administration of a further 10mg of obrisedronate on the basis of 20mg of atorvastatin resulted in an additional 50.3% reduction in LDL-C. A second patient study (TA-8995-06) showed a statistically significant decrease in Lp (a) levels after 12 weeks of olanzapine treatment.

Two phase 2 studies of olanzapine (TA-8995-303 and TA-8995-201) are currently nearly completed. The first study, TA-8995-303, was evaluating the LDL lowering effect of olanexidine 5mg in combination with ezetimibe 10mg in a participant with mild dyslipidemia. A second study, TA-8995-201, is evaluating the LDL lowering effect of olanzapine (both 5mg and 10mg doses) as an adjunct to high intensity statin therapy in participants with dyslipidemia who received high intensity statin therapy. Two phase 3 studies of 10mg of obiprap are currently underway, which investigate the treatment of elevated LDL-C levels in participants who have been diagnosed with atherosclerotic CV disease (ASCVD) as well as heterozygous familial hypercholesterolemia. One study (TA-8995-301) will incorporate participants receiving maximal tolerogenic lipid-lowering therapy, including statin doses at maximum tolerogenic doses of LDL-C incorporation standard >100 mg/dL. Another study (TA-8995-302) will also incorporate participants who are receiving maximum tolerated lipid lowering therapy, which includes statin doses at maximum tolerated doses of LDL-C inclusion criteria of > 70mg/dL and <100 mg/dL. These two key studies will together evaluate the effect of 10mg of obrisedronate in a population that needs to further reduce LDL-C in a range of baseline LDL-C values associated with current clinical practice.

The third phase 3 study (TA-8995-304) will investigate the effect of 10mg of obsemipirtine on clinical outcome (i.e., major adverse CV events including CV death, non-fatal myocardial infarction, non-fatal stroke, or non-selective coronary revascularization).

1.4 Basic principle

Chronic elevated LDL-C results in the progressive accumulation of atherosclerotic lesions requiring long-term treatment. Although lifestyle changes are the primary intervention, these rarely reduce plasma LDL-C by more than 15% and require medication to adequately treat hyperlipidemia.

Statin drugs are considered first line therapies that lower LDL-C levels. However, despite receiving lipid lowering therapies with statins, many patients fail to achieve acceptable LDL-C levels.

Inhibitors of proprotein convertase subtilisin kexin type 9 (PCSK 9) are an alternative to statin drugs. However, these therapies have several limitations including high cost, limited long-term outcome data relative to statins, and muscle-related events. Furthermore, since PCSK9 inhibitors are injectable, this is a less attractive option for patients who prefer oral drugs.

Thus, the need for therapies that effectively reduce elevated LDL-C levels and CV risks at acceptable cost, convenient dosage forms, and more advantageous safety profiles to promote long term use and patient compliance remains unmet. Olanzapine has proven to be safe and effective in lowering LDL-C as an oral CETP inhibitor, among other beneficial effects. The combination of olanzapine with ezetimibe, an oral cholesterol absorption inhibitor, may be a valuable alternative to PCSK9 inhibitors for patients who, despite receiving high intensity statin therapy, still need additional LDL-C lowering.

1.4.1 Basic principles of the combination therapy of obiprifepin and ezetimibe

Ezetimibe selectively inhibits intestinal cholesterol absorption. The integrated analysis of 8 randomized, double-blind, placebo-controlled studies demonstrated that using ezetimibe as a monotherapy for hypercholesterolemic patients significantly reduced serum LDL-C levels, with a statistically significant 18.58% reduction in the mean of LDL-C compared to placebo. Ezetimibe is used in combination with statin therapy to further reduce LDL-C levels. A global analysis of 27 studies involving 21,000 patients showed a further 15.1% reduction in LDL-C in patients treated with the combination of statin and ezetimibe compared to statin alone. In the Improvement (IMPROVE) -IT study, a comparison of 40mg daily simvastatin (simvastatin) with 40mg of simvastatin + ezetimibe 10mg in a combination of 18,144 patients with acute coronary syndromes showed that the future CV event incidence in the combination administration group further achieved a moderate but statistically significant (2% decrease in absolute risk over 7 years) decrease compared to statin alone. These extensive long-term studies have also demonstrated that ezetimibe has excellent safety characteristics. Importantly, additional Mendelian randomization studies have revealed that CETP inhibitor HMGCR inhibitor interactions do not occur when the CETP inhibitor is used in combination with ezetimibe.

A phase 2 study is currently underway to evaluate the LDL lowering effect of 5mg of obrisetrapib in combination with 10mg of ezetimibe in subjects with mild dyslipidemia and prior to no CV events (TA-8995-303).

To assess the likelihood that the use of higher doses of obipratropium in combination with ezetimibe increased the reduction in LDL-C and ApoB, the study will assess the effect of the use of 10mg of obipratropium in combination with 10mg of ezetimibe as part of the drug regimen directed by the guidelines of the participants for the participants who are currently receiving high-intensity statin therapy (40 mg or 80mg of atorvastatin or 20mg or 40mg of rosuvastatin).

1.4.2 Dose selection rationale for obiprift

In clinical studies in healthy volunteers, single doses of olanzapine up to 150mg and multiple doses up to 25mg per day are generally well tolerated within 21 days. In clinical studies of patients, olanzapine is also well tolerated after a daily dosing of 10mg for 12 weeks, either alone or in combination with 2 different statins. Near maximum effect was observed in the 10mg dose of obiprift. At this dose level, CETP activity and concentration decreased, HDL-C levels increased, and LDL-C levels decreased. In any clinical study, dose-related Adverse Events (AEs) were not found, and clinically significant changes in vital signs, electrocardiography (ECG), or hematology or biochemistry parameters were not found. At a dose level of 10mg of obiprap, a statistically significant decrease in Lp (a) levels from baseline was also observed. Thus, the present study will use a 10mg dose of obsemitrapib for participants with ASCVD who have not obtained adequate control despite receiving maximum tolerogenic lipid-modulating therapy.

1.5 Risk/benefit

The primary pharmacology in vitro, ex vivo and in vivo studies has demonstrated that obrisetrapib has the ability to inhibit CETP, lower LDL-C levels, raise HDL-C levels, and importantly reduce the number of atherogenic ApoB-containing particles in a manner useful for the treatment of dyslipidemia.

Safety pharmacology studies have shown that in rats, doses of up to 300mg/kg of obsemipirimip have no adverse effect on key physiological systems (e.g., central nervous system, respiratory system, gastric emptying, urinary tract and steroid hormone production [ including aldosterone levels ]).

In clinical studies of patients, olanzapine is also well tolerated after a daily dosing of 10mg for 12 weeks, either alone or in combination with 2 different statins. In any clinical study, dose-related AEs were not identified, and clinically significant changes in vital signs, ECG, or hematology or biochemistry parameters have not been found.

2. Purpose of investigation

2.1 Main purpose(s)

The main objective of this study was to evaluate the effect of the combination therapy of olanzapine 10mg + ezetimibe 10mg on LDL-C on day 84 compared to placebo when used as an adjunct to high intensity statin therapy.

2.2 Secondary purposes

Secondary objectives of this study were ranked in order including the following:

● Assessing the effect of 10mg monotherapy of obrisetrapib on LDL-C on day 84 when used as an adjunct to high intensity statin treatment compared to placebo;

● Assessing the effect of the combination therapy of olanzapine 10mg + ezetimibe 10mg on ApoB on day 84 when used as an adjunct to high intensity statin therapy compared to placebo, and

● The effect of 10mg monotherapy of obrisetrapib on ApoB on day 84 was evaluated as compared to placebo when used as an adjunct to high intensity statin therapy.

2.3 Exploratory purposes

The exploratory purposes of this study included the following:

● Assessing the effect of the combination therapy of olanzapine 10mg + ezetimibe 10mg and the monotherapy of olanzapine 10mg on non-high density lipoprotein cholesterol (non-HDL-C), very low density lipoprotein cholesterol (VLDL-C), HDL-C, TG, apoE, lp (a) and HDL-ApoE on day 84 when used as an adjunct to high intensity statin therapy;

● Evaluating the effect of the combination therapy of olanzapine 10mg + ezetimibe 10mg and the monotherapy of olanzapine 10mg on the proportion of participants who reached a predetermined LDL-C goal on day 84 when used as an adjunct to high intensity statin therapy;

● Assessing the mean trough plasma levels of the combination therapy of 10mg of obipratropium + ezetimibe and 10mg of obipratropium monotherapy at steady state on days 28, 84 and 112, and

● Safety and tolerability characteristics of the olanzapine 10mg + ezetimibe 10mg combination therapy and the olanzapine 10mg monotherapy as adjunctive therapy to high-intensity statin therapy were evaluated as assessed by clinical laboratory values and incidence of AE.

3. Description of the study

3.1 Summary of study design

This study will be a placebo-controlled, double-blind, randomized, phase 2 study to evaluate the efficacy, safety and tolerability of 10mg of obrisedronate, whether used in combination with 10mg of ezetimibe or as monotherapy in adjunctive therapy as a high intensity statin therapy. This study will be conducted at about 20 sites in the united states.

3.1.1 Screening cycle

At the time of the screening visit, participants will be required to sign an Informed Consent Form (ICF) before any study-related procedures are performed. After signing the ICF, participants will be eligible for study.

3.1.2 Treatment cycles

Within 2 weeks after the screening visit, participants will return to the study site on day 1 (visit 2) and confirm study qualification before randomizing and starting treatment. Approximately 114 eligible participants (38 participants per treatment group) will be randomly assigned to 1 of the following treatment groups at a ratio of 1:1:1:

● Combination therapy, olanzapine 10mg + ezetimibe 10mg (administered as a tablet of 10mg olanzapine and a capsule of 10mg ezetimibe);

● The single therapy of the obiprift comprises 10mg of the obiprift (which is applied in the form of a tablet of 10mg of the obiprift and a placebo capsule), or comprises

● Placebo (administered in the form of 1 placebo tablet and 1 placebo capsule).

During a treatment period of 12 weeks, participants will orally administer the prescribed study medication once with water at about the same time every morning on days 1 through 84. Participants will return to the study site on day 28 (+ -2 days), day 84 (+ -2 days) and day 112 (+ -2 days) for efficacy, safety and Pharmacokinetic (PK) assessment. Subjects, researchers, clinical study organizations (CRO) and sponsors were blinded to all lipid results from day 1 (visit 2) of the first subject until all enrolled subjects completed the day 84 (end of treatment) visit or exited the study to protect blindness of treatment assignment.

3.1.3 Safe follow-up period

Participants will return to the study site for a safe follow-up (visit 5) approximately 4 weeks after the treatment cycle has ended

Thereby performing security and PK assessment.

3.1.4 Emergency measure for coronavirus diseases in 2019

In the limiting case of COVID-19, the researcher is responsible for ensuring the safety of the participants. The sponsor will implement and record the mitigation strategy if necessary. At the discretion of the researcher, research visits may be performed either at the clinic or in a virtual manner. If done in a virtual manner, the visit will include alternative methods of study drug safety, efficacy, and distribution/collection, including but not limited to phone/video contact, alternative locations of biological sample collection, alternative safe delivery of study drugs, home healthcare (if available), and transfer of participant data to and from the home healthcare service and the study site in a secure manner.

If such emergency actions occur, the sponsor will record the changes made and communicate advice regarding such changes to minimize or prevent interference with the study and to support the implementation of such changes at the study site. The documentation of these situations and the management of participants by the study site should be recorded in the study file of the researcher. Without being affected by COVID-19, researchers and participants are expected to follow prescribed protocol requirements.

3.2 Study indications

The indication for this study was dyslipidemia.

4 Participant selection and exit

4.1 Inclusion criteria

Participants meeting all of the following criteria will be eligible to participate in the study:

1. Knowing the study program, is willing to follow the study schedule and agree to participate in the study by signing written informed consent prior to the screening program;

2. screening men or women between 18 and 75 years of age (inclusive) at visit;

● Women may opt in if they meet all three criteria:

o is not pregnant;

O is not breast-feeding, and

The pregnancy was not intended during the study.

● Urine pregnancy test of women with fertility at screening visit must be

● Negative. Note that if the woman meets 1 of the following criteria recorded by the researcher,

● Then it is not considered to be fertility:

Hysterectomy or tubal ligation was performed for at least 1 cycle before signing the ICF, or

It is a postmenopausal female, defined as a female of age no less than 55 years old no less than 1 year from its last menstrual period, a female of age <55 years old no less than 1 year from its last menstrual period and Follicle Stimulating Hormone (FSH) levels in the postmenopausal range.

● Women with fertility must agree to use an effective method of avoiding pregnancy within 90 days from screening to the last follow-up. Within 90 days from screening to the last follow-up, men with fertility must agree to use an effective method of avoiding pregnancy. An effective method of avoiding pregnancy is to continue the correct use of contraceptive methods with a pearl index <1 (including implantable contraceptives, injectable contraceptives, oral contraceptives, percutaneous contraceptives, intrauterine devices, spermicidal contraceptive diaphragms, spermicidal male or female condoms or cervical caps) or fertility-free sexual partners.

3. Screening for fasting LDL-C levels >70mg/dL and TG levels <400mg/dL, and

4. Prior to screening, high-intensity statin therapy (40 mg or 80mg or 20mg or 40mg of rosuvastatin) is currently being received at a stable dose for 8 weeks and is intended to maintain the same stable dose throughout the duration of the study.

4.2 Exclusion criteria

Participants meeting any of the following criteria will be excluded and unable to participate in the study:

1. screening body mass index ≡40kg/m2 at visit;

2. CV diseases of current clinical significance include, but are not limited to:

● Major adverse CV events within 3 months prior to randomization, or

Note that the major adverse CV event is defined as CV death, non-fatal myocardial infarction, non-fatal stroke, or non-selective coronary revascularization.

● The new york heart association functional classification class III or IV heart failure.

3. Screening glycosylated hemoglobin (HbA 1 c) ≡10% at visit;

4. hypertension is not controlled, i.e. the sitting systolic pressure >160mmHg and/or the sitting diastolic pressure >90mmHg. Allowing a retest to be performed, wherein if the retest result is no longer exclusive, the participants can be randomly grouped;

5. Active muscle disease or persistent creatine kinase concentration >3 x upper normal value limit (ULN). A retest is allowed to verify the results after 1 week, at which time the participants may be randomly grouped if the retest results are no longer exclusive;

6. Approximate glomerular filtration rate <60mL/min calculated using the Chronic kidney disease epidemiological collaborative set of equations (Chronic KIDNEY DISEASE Epidemiology Collaboration equation);

7. Liver function abnormalities manifested by any laboratory abnormality of >2 XULN for gamma-glutamyl transferase, alanine aminotransferase or aspartate aminotransferase, or >1.5 XULN for total bilirubin;

8. Anemia, defined as male hemoglobin concentration <11g/dL, and female hemoglobin concentration <9g/dL;

9. a history of malignancy was present within 5 years of screening, except for non-melanoma skin cancers;

10. a history of alcohol and/or drug abuse within 5 years prior to screening;

11. treatment with other study products or devices within 30 days prior to screening or within 5 half-lives (whichever is longer);

12. a history of any clinical trial involving evaluation of obipratropium;

13. Treatment with any PCSK9 inhibitor within 10 weeks prior to randomization, or with bevacizidine within 2 weeks prior to randomization;

14. The researcher considers evidence of any other clinically significant non-cardiac disease or condition that would prevent the participation of the participant in the study, or

15. CETP inhibitors are known to be allergic or intolerant.

4.3 Retesting

If the investigator sees that the laboratory abnormality during the screening is transient, the laboratory test may be repeated once during the screening. The researchers performed a logical basis for randomization.

4.4 Rescreening

After consultation and approval by sponsors and/or medical inspectors, rescreening of participants who are not meeting the criteria for eligibility for the study after screening may be considered. The rescreened participant will be assigned a new participant number. Rescreening should be performed no less than 5 days after the last screening visit.

4.5 Exit Standard

Participation in this study may be terminated for any of the following reasons:

● Participants withdraw consent or are required to discontinue the study for any reason;

● Any therapeutic condition or situation that exposes the participant to significant risk and/or does not allow the participant to follow the regimen requirements occurs;

● Any Serious AE (SAE), clinically significant AE, serious laboratory abnormality, inter-onset or other therapeutic condition that indicates to the researcher to continue participation in a maximum benefit that is not in line with the subject;

● Pregnancy;

● The need to use concomitant medications that are disabled;

● Participants failing to follow the program requirements or study-related procedures, or

● The sponsor or regulatory agency terminates the study.

Unless the participants withdraw the consent, participants who had previously discontinued the study medication should be encouraged to immediately complete the complete panel (full panel of assessment) scheduled for the early termination visit (Early Termination Visit) and complete the safety follow-up (visit 5) 4 weeks after administration of the last dose of study medication. For those who had previously discontinued study medication without withdrawal of consent or those who have prematurely withdrawn the study, PK samples will not be collected during the safety follow-up. The reason for participant exit must be recorded in an electronic case report table (eCRF).

If a participant loses access (lost to follow-up), an attempt must be made to contact the participant and record in the participant's treatment record.

The exiting participant will not be replaced.

5 Study treatment

5.1 Treatment group

The participants will be randomly assigned to 1 of the following treatment groups at a ratio of 1:1:1:

5.2 Basic principle of quantitative administration

In clinical studies in healthy volunteers, single doses of olanzapine up to 150mg and multiple doses up to 25mg per day are generally well tolerated within 21 days. In clinical studies of patients, olanzapine is also well tolerated after a daily dosing of 10mg for 12 weeks, either alone or in combination with 2 different statins. Near maximum effect was observed in the 10mg dose of obiprift. At this dose level, CETP activity and concentration decreased, HDL-C levels increased, and LDL-C levels decreased. In any clinical study, dose-related AEs were not identified, and clinically significant changes in vital signs, ECG, or hematology or biochemistry parameters have not been found. At a dose level of 10mg of obiprap, a statistically significant decrease in Lp (a) levels from baseline was also observed. Thus, the present study will use a10 mg dose of obsemitrapib for participants with ASCVD who have not obtained adequate control despite receiving maximum tolerogenic lipid-modulating therapy. The ezetimibe dose of 10mg is the current FDA approved dose.

5.3 Randomization and blinding methods

Participants meeting all qualification criteria will be randomly assigned to the study. The participants will be randomized in a 1:1:1 ratio to receive either combination therapy (olanexidine 10mg + ezetimibe 10mg [ administered as a tablet of 10mg of olanexidine and a capsule of 10mg of ezetimibe ]), olanexidine monotherapy (olanexidine 10mg [ administered as a tablet of 10mg of olanexidine and a capsule of placebo ]), or placebo (administered as a tablet of 1 placebo and a capsule of 1 placebo).

At random grouping, participants will be classified into different classes according to their screening visit LDL-C levels (. Gtoreq.100 mg/dL or <100 mg/dL). Participants will be assigned to 1 out of 3 treatment groups using an automated Interactive Response Technology (IRT) system.

Participants, researchers, CRO and sponsors were blind to all lipid results from the first participant starting on day 1 (visit 2) until all enrolled participants completed the day 84 (end of treatment) visit or exited the study to protect blindness of treatment assignment.

5.5 Drug delivery

5.5.1 Formulations and packages

The study drug consisted of 10mg of obrisetrapib tablets or matched placebo tablets and over-encapsulated 10mg ezetimibe tablets or matched placebo capsules. All study drugs were manufactured according to the international consortium of coordination (International Council for Harmonisation, ICH) current good manufacturing practice (current Good Manufacturing Practice).

The obiprift tablet is a round white film coated tablet without identification mark and contains 10mg of obiprift calcium drug substance. Excipients present in the tablet core are microcrystalline cellulose, mannitol, sodium starch glycolate, colloidal silicon dioxide and magnesium stearate. A commercially available film coating formulation (Opadry II white (white), available from carrecon (Colorcon)) was applied to the core.

The placebo tablet of obrisetrapib is a matched round white film coated tablet without an identification mark. Excipients present in the tablet core are microcrystalline cellulose, mannitol, sodium starch glycolate, colloidal silicon dioxide and magnesium stearate. A commercially available film coating formulation (Opadry II white (white), available from carrecon (Colorcon)) was applied to the core.

Ezetimibe capsules are 10mg ezetimibe tablets enclosed in a capsule shell, 1 tablet per capsule. Each capsule also contains excipient materials commonly used for tablets as fillers to prevent the tablets from rattling in the capsule shell.

Placebo capsules matched to ezetimibe capsules are the same capsule shells filled with only the excipient fill material (no tablets).

The obrisetrapib and placebo tablets and ezetimibe and placebo capsules will be packaged in a kit providing 2 study drugs for each treatment group. These kits will be labeled with clear labels to indicate tablets/capsules for daily use. Each kit will provide a supply of sufficient 28 days of dosing, sufficient to provide an additional 4 days of dosing if the participant needs to delay the next visit. The shelf life will be specified based on the stability of the individual study drug and will not exceed the shelf life of the ezetimibe tablet dosed. The kit will be stored below 25 ℃.

Physical, chemical and pharmaceutical formulation properties and characteristics of the obiprift tablet are set forth in the researchers' manual. All study drugs will be labeled according to all applicable local regulatory requirements.

5.5.3 Study of drug administration

Study drugs (1 tablet and 1 capsule) will be administered orally with water by the participants, once a day at about the same time in the morning on days 1 through 84. On the day of the scheduled visit, study medication should be administered with water after all fasted blood samples are collected. At visit 3 and 4, the participants dosed the kits received from the previous visit (visit 2 and visit 3, respectively).

If a participant forgets to take a study medication on a certain day, he should take the next dose normally, rather than double the dose to compensate for the forgotten dose.

5.5.4 Treatment compliance

Compliance with the study drug regimen will be assessed by counting unused tablets and capsules. Participants will be instructed to bring all unused study medication to the study site at visit 3 and 4. During the treatment period, if compliance is not between 80% and 120% (inclusive), the participants will be counseled about the importance of compliance versus the regimen. If 2 consecutive visits exceed the limit, the researcher and sponsor will decide whether the participant should exit the study.

5.5.5 Storage and liability

All study medications must be stored in a safe area where only researchers and authorized study site personnel enter at temperatures below 25 ℃ (77°f). According to regulatory requirements, a researcher or designated research site personnel must record the amount of research medication dispensed and/or administered to the participant, the amount returned by the participant, and the amount received at and returned to the sponsor (or representative), if applicable. Throughout the course of the study, a study drug liability record must be kept. The responsibility of this study was in the form of tablets or capsules. The disagreement value (discrepancy) will be mediated or resolved. The final treatment procedure without study drug will be provided in the appropriate study manual.

5.6 Previous and concomitant drugs and/or procedures

5.6.1 Excluded drugs and/or procedures

Within 30 days or 5 half-lives (whichever is longer) prior to screening, subjects must not receive treatment with other study products or devices. The participants had to stop taking any PCSK9 inhibitor within 10 weeks prior to the random grouping, or to stop taking bevacizidine within 2 weeks prior to the random grouping.

5.6.2 Drugs and/or procedures allowed

Prior to screening, subjects must currently be receiving high-intensity statin therapy (atorvastatin 40mg or 80mg or rosuvastatin 20mg or 40 mg) at a stable dose for 8 weeks and intend to maintain the same stable dose throughout the duration of the study.

5.6.3 Record past and concomitant drug use

The drug used within 28 days prior to the screening visit will be recorded. Whether or not permitted according to the regimen, any drug administered in addition to the study drug must be recorded in the concomitant drug eCRF.

7. Efficacy and pharmacokinetic assessment

7.1 Efficacy assessment

The primary efficacy endpoint was the percent change in LDL-C from day1 to day 84 for the 10mg combination treatment of obrisedrotrapib 10mg + ezetimibe compared to the placebo group.

The secondary efficacy endpoints are arranged in rank order including the following:

● Percentage change in LDL-C from day 1 to day 84 for 10mg monotherapy with obiprifepin

● The treatment group was compared to the placebo group;

● Percentage of ApoB change of 10mg of obrisetrapib +10 mg of ezetimibe from day 1 to day 84

● Comparing the combined treatment group with the placebo group, and

● Percentage of ApoB change from day 1 to day 84 in the 10mg monotherapy treatment of obrisetrapib compared to the placebo group.

The exploratory efficacy endpoints included the following:

● Percentage of change in the non-HDL-C, VLDL-C, HDL-C, TG, apoE, lp (a) and HDL-ApoE of the 10mg combination treatment of olanzapine 10mg + ezetimibe 10mg compared to the placebo and the 10mg monotherapy treatment of olanzapine compared to the placebo, and

● The proportion of participants of LDL-C <2.6mmol/l (< 100 mg/dL), LDL-C <1.8mmol/l (< 70 mg/dL) and LDL-C <1.3mmol/l (< 50 mg/dL) was achieved on day 84 for the combined treatment group of 10mg of obreplacement for obreplacement and 10mg of ezetimibe for the monotherapy treatment group compared to the placebo group.

Blood samples of the lipid profile must be obtained under fasted conditions (i.e., after the participants have fasted for about 10 hours). For the purposes of this study, fasting will be defined as not feeding any substance via the mouth, except for water and any necessary medications. If the participants are not fasted, the investigator should reschedule the visit as soon as possible. Unless TG is 400mg/dL or LDL-C is 50mg/dL, the LDL-C level will be calculated using the Friedel's formula (FRIEDEWALD EQUATION), in both cases the LDL-C level will be measured directly by preparative ultracentrifugation (PREPARATIVE ULTRACENTRIFUGATION), also known as beta quantification (beta quantification). In addition, LDL-C will be measured for all participants at baseline (visit 2) and at the end of the 12 week treatment period (visit 4) by preparative ultracentrifugation, also known as beta quantification.

7.2 Pharmacokinetic assessment

Plasma obbesitrapine concentrations used in combination with ezetimibe and as monotherapy will be assessed at the scheduled PK collection times shown in appendix a. At the visit where dosing was scheduled (i.e., visit 2, visit 3, and visit 4), PK samples were collected using fasted laboratory samples (prior to dosing). At visit 3 and 4, the time between the last dose of study drug and PK sampling will be >24 hours. PK samples should be collected at about the same time as the previous visit following subsequent quantitative administration at visit 5.

8 Safety assessment

Safety and tolerability characteristics of 10mg of obbestrepide used in combination with 10mg of ezetimibe and as monotherapy will be assessed by clinical laboratory assessment (chemistry and hematology), vital signs, physical examination, and incidence of AE.

8.1 Adverse events

AE is defined as any unfortunate treatment event that occurs after administration of a drug product by a clinical study participant, which is not necessarily causally related to this treatment. Thus, an AE may be any adverse and/or unplanned sign (including abnormal laboratory findings), symptom, or disease that is temporally associated with (whether or not associated with) a pharmaceutical product under investigation. All AEs (including observed or proactively posed problems, complaints or symptoms) were recorded on the appropriate eCRF.

AE, including clinical laboratory rated variables, will be monitored and recorded starting from the time of the first dose of study treatment until the end of visit 5. The participants should be instructed to report any AEs they experience to the investigator, whether or not they consider the event to be caused by the study drug. Beginning on the day of the first dose receiving study treatment, the investigator should evaluate AE at each visit and record the event in the appropriate AE eCRF.

The researcher should identify as much as possible a specific disease or syndrome rather than individual related signs and symptoms and record it on eCFs. However, if the observed or reported signs or symptoms are not considered by the researcher as part of a particular disease or syndrome, they should be recorded in the eCRF as a separate AE. Furthermore, the situation that leads to the performance of a medical or surgical operation (e.g., surgery, endoscopy, tooth extraction or blood transfusion) should be recorded as AE rather than the operation itself.

Any treatment condition that has occurred on the day of the first dose of study treatment should be recorded as a medical history and not reported as AE unless the treatment condition or sign or symptom that occurred at baseline changes in severity, frequency or severity at any time during the study. In this case, it should be reported as AE.

As described below, clinically significant abnormal laboratory or other examination findings (e.g., ECG) detected during the study or occurring on the day of the first dose receiving study treatment and significantly worsening during the study should be reported as AEs. Researchers will use their medical and scientific judgment to decide whether or not abnormal laboratory findings or other abnormal assessments are of clinical significance. Abnormal laboratory values of clinical significance that occur during the study will be monitored until the repeated test returns to normal, stable or no longer of clinical significance. The abnormal test result determined to be erroneous should not be reported as AE. Laboratory abnormalities or other abnormal clinical findings (e.g., ECG abnormalities) should be reported as AEs if any of the following applies:

● If intervention is required due to an anomaly;

● If the study drug is required to take measures due to abnormality, or

● Based on the clinical judgment of the investigator.

8.1.1 Adverse (drug) reactions

All adverse and unintended reactions to the pharmaceutical product associated with any dose should be considered adverse drug reactions. "response" to a pharmaceutical product means the possibility that the causal relationship between the pharmaceutical product and AE is at least reasonable, i.e. such relationship cannot be excluded.

8.1.2 Unexpected adverse drug reactions

Unexpected adverse drug reactions are defined as adverse reactions that are inconsistent in nature or severity with applicable product information.

8.1.3 Assessment of adverse events by researchers

Researchers rate the severity (intensity) of each AE as mild, moderate or severe, and will also classify each AE with a "yes" or "no" category for its potential relationship to study medication.

Severity assessment

Mild-events that are easily tolerated and do not normally interfere with normal daily activities.

Moderate-an uncomfortable event sufficient to interfere with normal daily activities.

Severe-an event that disables work or performs the ability of normal daily activities.

Causal relationship assessment

The relationship of AE to study drug administration will be assessed according to the following definition:

Whether (irrelevant, unlikely relevant) -the time course between administration of study drug and AE occurrence or exacerbation excludes causal relationships and suspects the existence of other causes (concomitant drugs, therapies, complications, etc.).

It is (likely, probably or clearly relevant) -the time course between administration of study drug and occurrence or exacerbation of AE is causally related and other causes (concomitant drugs, therapies, complications, etc.) cannot be identified.

The definition implies a reasonable probability of causal relationships between events and study drugs. This means that there is a fact (evidence) or a arguments to indicate causal relationships.

The following factors should also be considered:

● Time sequence since study drug administration

The event should occur after administration of the study drug. The length of time from exposure to study drug to event should be assessed according to the clinical condition of the event.

● Potential, concomitant, intermittent disease

Each report should be evaluated based on the natural history and course of the disease being treated and any other disease that the participant may have.

● Concomitant drug-

Other medications that the participant is taking or treatments that the participant receives should be examined to determine if any of them can be considered to cause a related event.

● Known reaction forms of such research drugs

Clinical and/or preclinical data may indicate whether a particular response is likely to be a class effect (CLASS EFFECT).

● Exposure to physical and/or mental stress

Exposure to pressure may induce adverse changes in the recipient and provide a logical and better explanation for the event.

● Study of pharmacology and PK of drugs

Consideration should be given to studying the known pharmacological properties (absorption, distribution, metabolism and excretion) of the drug.

8.2 Serious adverse events

An AE or adverse reaction is considered to be a severe AE or adverse reaction if the researcher or sponsor deems it to lead to any of the following:

● Death;

● AE, which is dangerous and life-threatening;

Note that an AE or adverse reaction is considered "dangerous and life" if it is considered by the researcher or sponsor that it would be the occurrence of the AE or adverse reaction that would expose the participant to a direct risk of death. It does not include events that could lead to death if they occur in a more severe form.

● Hospitalization is required or the existing hospitalization time is prolonged;

note that any admission stay for at least 1 night will be considered a hospitalization (INPATIENT HOSPITALIZATION). According to this standard, an emergency room or emergency care visit that is not admitted will not be recorded as an SAE, a scheduled or planned surgical hospitalization prior to the informed consent being signed, or a chronotherapy (ELECTIVE TREATMENT) for an existing condition that is not worsened from baseline will not be recorded as an SAE. However, the unexpected complications and/or prolonged hospitalization occurring during the perioperative procedure should be recorded as AE and assessed for severity. Admission for social or situational reasons (i.e., no place to live, too far to get to hospital for a doctor's care, wheezing) will not be considered hospitalized.

● Sustained or severe disability/disability or ability to perform normal life functions is severely compromised;

● Congenital anomalies/birth defects, or

● Important treatment events.

Note that an important therapeutic event that does not meet any of the criteria described above may be considered an SAE when, based on appropriate medical judgment, it may be dangerous to the participants and may require medical or surgical intervention to prevent 1 of the results listed above. Examples of such therapeutic events include allergic bronchospasms requiring intensive treatment in an emergency room or home, blood cachexia or tics that do not result in hospitalization, or the development of drug dependence.

9. Statistical information

9.1 Analysis population

The Intent-to-Treat (ITT) population will include all participants randomly assigned to the study. The treatment classification will be based on randomized treatments.

The Modified ITT (mtt) population will include all participants in the ITT population who received at least 1 dose of any study drug and have baseline values for LDL-C assessment. Any efficacy measurements obtained during a safety follow-up (visit 5) performed after the participants permanently stopped study drug or after the participants received an excluded drug and/or procedure were removed from the mITT analysis. The treatment classification will be based on randomized treatments. The mtt population will be used for the primary analysis of all efficacy endpoints.

A study protocol (PP) compliant population will include all participants in the mITT population who have baseline values for LDL-C assessment, have day 84 values for LDL-C assessment, and do not develop significant study protocol bias that could affect the primary efficacy endpoint. PP populations and exclusion reasons will be finalized before study blinding.

The PK population will include all participants in the mITT population who collected enough blood samples for effective approximation of PK parameters.

The safety population will include all participants who receive at least 1 dose of any study medication. The treatment classification will be based on the actual treatments received. The security population will be the main population for security analysis.

10 Results

The highest results of the clinical study are summarized in tables 14.1.1.1 to 14.3.1.1 below.

Arrangement of table 14.1.1.1 for all random grouping participants

Table 14.1.4.1 ditt population demographics and baseline characteristics

TABLE 14.2.1.1.1MITT population LDL-C (mg/dL) summary

TABLE 14.2.1.1.1a summary of LDL-C (mg/dL) of exploratory populations

TABLE 14.2.2.2.1 MITT population of ApoB (mg/dL) summaries

Table 14.2.2.1 summary of ApoB (mg/dL) of exploratory population

TABLE 14.2.8.1MITT population summary of Lp (a) (nmol/L)

Table 14.2.8.1 summary of Lp (a) (nmol/L) for exploratory population

Table 14.3.1.1 adverse event overview of security population

Example 11

Synthesis of amorphous obiprift calcium

The examples in this section are provided by way of illustration and not limitation. The described embodiments may represent only some embodiments and it should be understood that the following examples are illustrative and not limiting. Unless otherwise indicated, all substituents are as previously defined. Reagents and starting materials are readily available to one of ordinary skill in the art. The specific synthetic steps of each of the routes may be combined in different ways, or in combination with steps of different schemes, to produce the compounds described herein.

Scheme 1

Referring to scheme 1, amorphous obsemicalcium (compound 3) was prepared by six chemical steps and three mesylate salts isolated from (2 r,4 s) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1A), tert-butyl-4- (2-chloropyrimidin-5-yloxy) -butyrate (compound 1B) and 3, 5-bis (trifluoromethyl) benzyl bromide (compound 1E). Compound 1A was coupled with compound 1B by a palladium catalyzed reaction to produce a solution of (2 r,4 s) -4- [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1C), which was not isolated but reacted directly with excess ethyl chloroformate in the presence of pyridine to produce (2 r,4 s) -4- [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate, which was isolated as crystalline methanesulfonate (compound 1D). The crystalline methanesulfonate salt, compound 1D, was alkylated with 3,5 bis (trifluoromethyl) benzyl bromide (compound 1E) under strongly basic conditions to give a solution of (2 r,4 s) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1F) in toluene. Compound IF is then subjected to acidic cleavage of tert-butyl ester to yield a solution of ethyl (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (Compound 1). Compound 1 is then converted to compound 2, which is a solvate of (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 2). Finally, compound 2 is converted to amorphous hemicalcium salt (compound 3) and milled to the target particle size. Compound 2 is crystalline obiprift HCl and compound 3 is amorphous obiprift hemi-calcium. The FT-IR spectrum of the milled amorphous obiprift hemi-calcium is seen in fig. 52. The solution state ¹ H-NMR spectrum consistent with the chemical structure of the calcium obrisedronate is seen in FIG. 53.

Each step in the preparation of (2 r,4 s) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1), intermediate HCl intermediate (compound 2) and the corresponding amorphous calcium salt (compound 3) will be described in more detail in examples 11.1 to 11.16 below.

Examples 11.1 to 11.3, 11.5, 11.7, 11.9, 11.11 to 11.12 illustrate methods of manufacturing steps in the process of preparing amorphous obiprift semi-calcium (compound 3), and examples 11.4, 11.6, 11.8, 11.10 and 11.13 provide additional methods of preparing the indicated compounds. The methods in these examples sometimes represent more than one batch prepared from the indicated compounds produced.

Examples 11.14 to 11.15 illustrate the milling of amorphous obiprift calcium (compound 3) and example 11.16 illustrates a process for preparing crystalline obiprift calcium.

EXAMPLE 11.1 preparation of (2R, 4S) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (Compound 1A free base)

(2R, 4S) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (Compound 1A) (62 kg,182mol,1.00 eq.) was added together with toluene (375L) to a reaction vessel equipped with a reflux condenser. The resulting slurry was stirred at 52 ℃ and 1M aqueous sodium hydroxide solution (322 l,5.2 vol.) was added. The reaction mixture was stirred until all solids were dissolved and then cooled to 20 ℃. Stirring was stopped and the reaction mixture was allowed to separate into two phases. The bottom aqueous phase was drained and an aqueous sodium chloride solution (310 l,5.0 vol.) was added. The reaction mixture was then stirred at 20 ℃ for 30 minutes. Stirring was again stopped and the reaction mixture was allowed to separate into two phases. The bottom aqueous phase was drained and deionized water (310 l,5.0 vol.) was added. The reaction mixture was then stirred at 20 ℃ for 30 minutes. Stirring was again stopped and the reaction mixture was allowed to separate into two phases. The bottom aqueous phase was separated. The resulting organic solution is then subjected to vacuum distillation at an internal temperature of 65 ℃ or less. Distillation was continued until a final visual volume of 4.0 volumes (250L) was reached. The reaction vessel was then cooled to 20 ℃ to provide a solution of (2 r,4 s) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1A-free base) in toluene, with a small amount of water present. Compound 1A-free base was not isolated but was used directly in example 11.2.

EXAMPLE 11.2 preparation of (2R, 4S) -4- [5- (3-tert-Butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1C)

Additional toluene (107 l,1.5 vol.) was added to a reaction vessel ("vessel a") containing (2 r,4 s) -4-amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1A-free base) in toluene from the previous step, wherein water was <1000ppm. Tert-butyl-4- (2-chloropyrimidin-5-yloxy) -butyrate (compound 1B) (54.6 kg,200mol,1.10 eq.) was then added to vessel a along with tert-butanol (t-BuOH) (122 l,1.55 vol.). The reaction mixture was stirred and nitrogen was vented. Simultaneously, palladium acetate (410 g,1.8mol,1 mol%) was added to a second reaction vessel ("vessel B") under nitrogen. (S) -BINAP (2.48 kg,4.0mol,2.2 mol%) and toluene (107L, 1.5 vol.) were further added to vessel B, and the resulting mixture was stirred to form a red/orange Pd-BINAP solution. The orange/red Pd-BINAP solution of reaction vessel B was transferred to vessel A. K ₃PO₄ (85 kg,400mol,2.20 eq.) was further added to vessel A and the resulting reaction mixture was heated to an internal temperature of 72℃and stirred for at least 2 hours. The mixture was then cooled to 20 ℃, deionized water (124L) was carefully added, and the mixture was stirred for 30 minutes. The stirring was then stopped and the layers allowed to separate into two phases. The bottom aqueous phase was separated and 1M aqueous HCl (123L) was added with stirring. After 30 minutes, stirring was stopped again and the layers allowed to separate into two phases. The bottom aqueous phase was separated off and an aqueous sodium chloride solution (326 kg,5.26 vol.) was added while stirring. After 30 minutes, stirring was stopped again and the layers allowed to separate into two phases. The bottom aqueous phase was separated and deionized water (248 l,4.0 vol.) was added with stirring. after 30 minutes, stirring was stopped again and the layers allowed to separate into two phases. The bottom aqueous phase was separated. The resulting reaction mixture was then treated with ethylenediamine (1.60 kg,0.15 eq.) and stirred at 20 ℃ for 80 minutes. The reaction mixture was then filtered on a charcoal cartridge (charcoal cartridge) and the filtrate was returned to the clean vessel. The mixture is then distilled under partial vacuum (partial vacuum) at an internal temperature of 60 ℃ or less. Distillation was continued until a remaining approximately 2.50 volumes were visually observed in the reactor (155L), then acetonitrile (390L, 5.0 vol.) was added. The mixture is then vacuum distilled at an internal temperature of 60 ℃ or less. Distillation was continued until the presence of approximately 2.50 volumes was visually observed in the reactor (155L), and then the contents were cooled to 20 ℃. The reaction vessel was then filled with acetonitrile (390L, 5.0 vol.) until 11 volumes (about 620L) were reached by visual inspection, thereby obtaining (2 r,4 s) -4- [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1C) dissolved in acetonitrile. compound 1C was not isolated but was used directly in example 11.3.

EXAMPLE 11.3 preparation of (2R, 4S) -4- [5- (3-tert-Butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester in crystalline methanesulfonate form (compound 1D)

(2R, 4S) -4- [5- (3-t-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline (compound 1C) in acetonitrile (about 620L) was cooled to an internal temperature of <10℃and pyridine (72L, 900mol,4.9 eq.) was added. Ethyl chloroformate (136 l,1428mol,7.84 eq) was then added via the addition funnel while maintaining the internal temperature of the reactor contents at <10 ℃. The internal temperature of the reaction mixture was then increased linearly to 20 ℃ over the course of 3.5 hours. The mixture is then vacuum distilled at an internal temperature of 60 ℃ or less. Distillation was continued until approximately 2.50 volumes (155L) were visually present. Isopropyl acetate (471L, 6.6 vol.) is then added to the reaction vessel and distillation is continued under vacuum at an internal temperature of 60 ℃ or less until approximately 2.50 volumes (155L) remain visually. Isopropyl acetate (471 l,6.6 vol.), 1M hydrochloric acid (307 l,5.0 vol.) and 26% aqueous sodium chloride solution (63 l,1.2 vol.). Were then added to the reaction vessel. The resulting mixture was stirred for 30 minutes and then separated into two phases. The bottom aqueous phase was separated and saturated aqueous sodium bicarbonate (132 l,2.3 vol.) was added. The resulting mixture was stirred for 30 minutes and then separated into two phases. The bottom aqueous phase was separated and the remaining mixture was distilled under vacuum at 60 ℃ or less to give a total volume (250L) of approximately 4.0 volumes visually, thereby obtaining ethyl (2 r,4 s) -4- [5- (3-t-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (corresponding to the free base of compound 1D) in isopropyl acetate based on the weight of the solution.

Additional isopropyl acetate (86 l,1.4 vol.) and methyl tert-butyl ether (MTBE, 593l,9.6 vol.) were added to isopropyl acetate (to the free base of the corresponding compound 1D) and the jacket (jack) temperature was set to 20 ℃. Methanesulfonic acid (MsOH, 17.6kg, 1.0 equivalent based on the mmol of the compound (corresponding free base of compound 1D) was then added to the reaction mixture over 60 minutes. The resulting slurry was then stirred for 8 hours. The slurry was then vacuum filtered at 20 ℃. The solid cake was then washed with 75/25v/v isopropyl acetate (78 l,1.1 vol.) and methyl tert-butyl ether solution (236 l,2.8 vol.) and then dried under vacuum at 20 ℃ to obtain isolated ethyl (2 r,4 s) -4- [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (compound 1D) as crystalline methanesulfonate in 74% yield based on the number of moles of compound 1A. The purity of the crystalline compound 1D obtained was >99%.

Example 11.4 additional preparation of Compound 1C and Compound 1D

The following preparation is generally used to prepare multiple batches of compound 1C and compound 1D. For example, in some preparations, seeding was performed with compound 1D, while this was performed without other compounds, as discussed further below.

Pd (OAc) ₂ and (S) -BINAP were dissolved in toluene and stirred to form the corresponding Pd-BINAP complex (color changed to red) ("catalyst solution"). Toluene, compound 1B, compound 1A and K ₃PO₄ were added to the reactor and stirred. The target content of water was about 6%. The catalyst solution was added to the reactor mixture and the reaction mixture was heated to 70 ℃ to 75 ℃ with stirring.

After washing with HCl, brine and water, phase separation was then performed, EDA and toluene were charged, and the solution was stirred for about 90 minutes. The solution was passed through a cartridge containing activated carbon (Bei Geluo company (Begerow), F-9120) to remove palladium. After that, the solvent was switched from toluene to acetonitrile (MeCN) by distillation to obtain compound 1C.

Pyridine was added to the compound 1C solution and cooled to below 10 ℃ prior to the addition of ethyl chloroformate. Ethyl chloroformate was added portionwise to the compound 1C solution in one or two portions while controlling the temperature to NMT 10 ℃. The reaction mixture was then stirred at 17 ℃ to 27 ℃ for about 1 hour, thereby converting compound 1C to the free base of compound 1D (compound 1D-FB). Solvent conversion from acetonitrile to isopropyl acetate (iPrOAc) was accomplished by distillation, and the organic phase was washed with HCl (1M), brine, and aqueous NaHCO ₃ (NaOH may also be used), followed by reduction in volume by distillation.

Methanesulfonic acid (MsOH) and MTBE were added to a solution of compound 1D-FB in iPrOAc, and stirred. In some cases, seed crystals of compound 1D previously produced were added, but this is not essential. Crystallization of compound 1D then occurs, whether or not seed crystals are added. The solid product was filtered, washed with MTBE/iPrOAc (75/25) and dried on the filter.

EXAMPLE 11.5 preparation of (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1F)

(2R, 4S) -4- [5- (3-t-Butoxycarbonylpropoxy) pyrimidin-2-yl) ] amino-2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1D) (42 kg) and toluene (460 kg,12.7 vol.) in the form of crystalline methanesulfonate were added to a reaction vessel at a temperature of 5 ℃. Tetrabutylammonium bisulfate (3.5 kg,0.16 eq) and sodium tert-amyl alcohol (34.5 kg,4.8 eq) were then added and the resulting reaction mixture was stirred for 10 minutes and degassed with nitrogen. 3, 5-bis (trifluoromethyl) benzyl bromide (compound 1E) (28 kg,1.41 eq.) was then added to the reaction mixture and stirring was continued for 6.5 hours at 5 ℃. The reaction mixture was then treated with 1N acetic acid solution (320 kg) and allowed to stir at 20 ℃ for about 30 minutes. After this time, stirring was stopped and the mixture was allowed to separate into two phases. The lower aqueous phase was discarded and the reaction mixture was concentrated in vacuo at an internal temperature of 60 ℃ or less until about 3.3 volumes (137L) remained, thereby obtaining 36.8 weight percent of ethyl (2 r,4 s) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-t-butoxycarbonylpropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (compound 1F) in toluene in 97% yield based on the weight of the solution based on the moles of compound 1D.

Example 11.6 additional preparation of Compound 1F

Compound 1E was charged into a toluene solution containing compound 1D and tetrabutylammonium bisulfate. While cooling, sodium tert-amyl alcohol in toluene was added. The resulting reaction mixture was quenched with diluted acetic acid. The aqueous layer was separated and the product in the toluene layer was treated with charcoal and concentrated in vacuo (Compound 1F.)

EXAMPLE 11.7- (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1)

37Wt.% of ethyl (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-tert-butoxycarbonylpropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (compound 1F) in toluene (128.4 kg of a 37wt.% solution, corresponding to 47.5kg of compound 1F) was diluted to 32wt.% with additional toluene and then mixed with acetic acid (253 kg,5.33 wt.) and 6M HCl (109.9 kg,2.32wt, prepared in-situ with 66.1kg of concentrated HCl and 43.8kg of water). The resulting reaction mixture was stirred vigorously and heated to 48 ℃ for 3 hours. The reaction mixture was then cooled to 21 ℃ and then n-heptane (159.8 kg,3.36 wt.), acetonitrile (73.8 kg,1.55 wt.), and water (170 kg,3.58 wt.) were added. The resulting mixture was stirred for 34 minutes and then allowed to separate into two phases. The lower aqueous phase was then further treated with water (90 kg,1.89 wt.), n-heptane (95 kg,2.00 wt.), acetonitrile (38 kg,0.80 wt.) and toluene (42 kg,0.88 wt.) and stirred for a further 20 minutes before the organic phase was separated and the lower aqueous phase was discharged. the combined organic phases were then treated with water (240 kg,5.05 wt.) and stirred for an additional 30 minutes before separating into two phases. The lower aqueous phase was discarded and the upper organic phase was treated with 5% w/w sodium citrate tribasic dihydrate (34 kg,0.72 wt.) and water (205 kg,4.32 wt.). The resulting mixture was vigorously stirred for 30 minutes and then allowed to separate into two phases before discarding the lower aqueous phase. The remaining organic phase was treated again with water (240 kg,5.05 wt.) and stirred for 30 minutes before separating it into two phases and discharging the lower aqueous phase. The organic phase was then concentrated in vacuo to about 3 volumes (about 149L) with the internal temperature maintained at 50 ℃ or less. The reaction mixture was diluted with cyclopentylmethyl ether (CPME, 250kg,5.26 wt.) and stirred. The solution was then concentrated in vacuo to about 3 volumes (about 165L) with the internal temperature maintained at 50 ℃ or less. CPME (250 kg,5.26 wt.) was then added and the mixture was concentrated in vacuo to about 2.5 volumes (about 124L) with an internal temperature of 50℃or less to obtain 33.7 weight percent of (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1, free base form) in cyclopentylmethyl ether (CMPE) having 1 weight percent toluene based on the weight of the solution Less than 1 weight percent n-heptane.

Example 11.8 additional preparation of Compound 1

Compound 1F (solution in toluene) was mixed with acetic acid and 6M aqueous HCl. The biphasic mixture was vigorously stirred at 45 ℃ to 50 ℃ and then cooled to 20 ℃. After addition of water, acetonitrile and n-heptane, the mixture was extracted and the layers were separated.

The aqueous layer of the first extraction was diluted with water and the second extraction was performed with acetonitrile, n-heptane and toluene. The two organic extracts obtained were combined. The organic phase was washed with water and a 5% sodium citrate solution was added to bring the pH to 3.5 or more. The organic layer was washed with water and treated with activated carbon. Solvent conversion from toluene and n-heptane to CPME was performed by repeating vacuum distillation and packing with CPME, thereby obtaining compound 1.

EXAMPLE 11.9- (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester hydrochloride (compound 2)

33.7 Weight percent of (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1, free base form, 115.6kg,59.2 mol) in cyclopentyl methyl ether (CPME) from the previous step was added under nitrogen to a clean reaction vessel at a jacket temperature of 22 ℃. After dilution with CPME (27.8 kg/0.58 wt.), n-heptane (54.8 kg,1.15 wt.) was then added and the internal reaction temperature was raised to 39 ℃. 3.0M HCl in CPME (17.6 kg,0.37 wt.) was then added at a constant rate while maintaining the internal reaction temperature at 39 ℃. After HCl addition was complete, the internal temperature was raised to 52 ℃. Additional n-heptane (133.2 kg,2.80 wt.) was then added at a constant rate while maintaining the internal reaction temperature at 51 ℃. The reaction mixture was heated to 55 ℃ and then cooled to 49 ℃. An aliquot of the reaction mixture was taken and cooled to 11 ℃ at a linear cooling rate until a slurry (referred to herein as a "seed slurry") was formed containing crystals of compound 2 in CPME/n-heptane. A seed slurry of compound 2 (169 g,0.43 weight percent) in CPME/n-heptane was then added at 49 ℃ and this temperature was maintained for 105 minutes. The opaque reaction mixture was then cooled to 11 ℃ over the course of 12 hours at a linear cooling rate. The reaction mixture was then vacuum filtered at 11 ℃ to collect the solid wet HCl intermediate (compound 2). A mixture of CPME and n-heptane (56.6kg CPME,179kg n-heptane) was then added to the reaction vessel and cooled to 11 ℃. Half of the mixture was then poured through a filter drier and washed as a chromatograph (chromatography wash). The other half was washed as slurry through a filter (slurry wash). Compound 2 was not removed from the filter-drier, but was further purified by recrystallization according to the following procedure.

Compound 2 in cyclopentylmethyl ether (CPME) (77.6 kg) was added to a filter dryer containing compound 2 and heated to 25 ℃. The dissolved compound 2 was then transferred under nitrogen to a reaction vessel set to a reactor jacket temperature of 25 ℃ and the internal temperature was raised to 38 ℃. 3.1M HCl in CPME (6.4 kg) was added such that a total of 1.07 equivalents of HCl was obtained based on the determination of compound 1 in the crude product of compound 2 and the determination of HCl in the crude product of compound 2. N-heptane (139.4 kg) was then added and the internal reaction temperature was raised to 51 ℃. A seed slurry of compound 2 (29 g,0.87 weight percent) in CPME/n-heptane was then added at 50 ℃ and this temperature was maintained for 105 minutes. The opaque reaction slurry was then cooled to 11 ℃ over 12 hours at a linear cooling rate. The slurry was then vacuum filtered using a filter dryer at 9 ℃. 20vol.% CPME in n-heptane (57.4kg CPME,180kg n-heptane) was then added to the reaction vessel and cooled to 11 ℃. Half of the mixture was then poured through a filter dryer and washed as a chromatograph. The other half was washed as slurry through a filter dryer. The wet cake was then vacuum dried in multiple steps of jacket temperature 25 ℃, 35 ℃, 46 ℃, 54 ℃, to provide compound 2 with purity of 99.6 area% in 64% yield (from compound 1F) and residual solvent 0.3% w CPME and <0.1% w n-heptane.

Example 11.10 additional preparation of Compound 2

Compound 1 in solution was further diluted with n-heptane and heated to 40 ℃. At this temperature, about 3M HCl (1.1 meq relative to compound 1F) in CPME was added. The solution was further heated to 48 ℃ to 53 ℃ and charged with a second portion of n-heptane. The clear solution was cooled to 53 ℃ for seeding with compound 2 seed crystals optionally (seeding is optional but preferred in the manufacturing environment). If seeding is performed, the solution is desaturated at 53 ℃ and then cooled to 10 ℃ over 12 hours.

A crystal solidification procedure (repeated heating and cooling cycle procedure) may be performed to improve the color of the resulting filtered compound 2. The product suspension was filtered, washed once with cooled CPME/n-heptane (20:80 vol), and once with cooled n-heptane, and then vacuum dried.

EXAMPLE 11 Ethyl (2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylate (compound 3)

(2R, 4S) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester hydrochloride (Compound 2,35.0kg,48.4 mol) was added to isopropyl acetate (IPAC, 214kg,6.11 wt.) in an inert reactor and stirred at 22℃to achieve dissolution. Deionized water (245 kg,7.00 wt.) was added, the reaction mixture was stirred at 23 ℃ for 35 minutes, then stirring was stopped, the phases separated, and the lower aqueous phase was removed. The process of adding deionized water (245 kg,7 wt.), stirring and removing the lower aqueous phase was repeated 3 more times. The organic phase was then concentrated to about 71L (about 2 vol.) under reduced pressure while maintaining the internal temperature at 55 ℃ or less. Ethanol (115 kg,3.29 wt.) was then added, and the reaction mixture was concentrated to about 78L (about 2 vol.) under reduced pressure while maintaining the internal temperature at 55 ℃ or less. The process of adding ethanol (115 kg,3.29 wt.) and concentrating was repeated two more times. The reaction mixture was then cooled to 25 ℃ and charcoal treated through the cartridge. The cartridge was then rinsed with ethanol (100 kg,2.86 wt.) and concentrated in vacuo to 147L (about 3.8 vol.) at 55 ℃ or less, followed by the addition of 35L of EtOH (1.0 vol.) to afford (2 r,4 s) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 1) as the free base in ethanol. Then, a 14wt.% NaOH solution (15.8 kg,1.13 eq.) was added to the reaction vessel containing compound 1 in ethanol with the reaction temperature maintained at 20 ℃. The reaction mixture was stirred at 20 ℃ for 5 hours to achieve complete conversion.

34% Wt. calcium chloride (aqueous) (10.8 kg) was added to the inert reactor. Deionized water (336L, 9.61wt. with respect to compound 1) and ethyl acetate (15 kg, 0.43wt. with respect to compound 1) were then added and the mixture was stirred for 30 minutes to provide "solution B".

Solution B was then cooled to 9 ℃ while stirring. Solution a (see above) was then added to solution B via the filter over 90 minutes with the temperature maintained at 10 ℃. The solution A vessel was then rinsed forward into solution B with additional ethanol (50 kg, 1.43wt. relative to compound 1). The resulting slurry was stirred at 9 ℃ for 1 hour. The solids were then collected by filtration and rinsed with deionized water (2 x 175kg, 5wt. relative to compound 1). The solid was then dried under vacuum at 50 ℃ for 21 hours to obtain 27.6kg of amorphous obiprift calcium (compound 3) with <1 weight percent water (77% yield based on moles of compound 2). Compound 3 was reprocessed as described below in example 11.12.

EXAMPLE 11.12 reprocessing of Compound 3

Compound 3 (27.6 kg) was dissolved in ethanol (55.2 kg, 2wt. relative to compound 3) at 45 ℃ to 48 ℃ and subsequently cooled to 11 ℃. The solution was filtered into a pre-cooled (about 10 ℃) mixture consisting of CaCl ₂ aqueous solution (8.2 kg of 33 to 35 weight percent, 0.3 wt.), water (262 kg,9.5 wt.) and ethyl acetate (12.6 kg,0.46 wt.). The resulting suspension was filtered off and washed with water (2 x 5wt.,138 kg/washing step) and the solid was dried in vacuo for 23 hours while maintaining the internal temperature at 45 ℃ or less to obtain 24.8kg (91% yield) of amorphous hemicalcium salt with <1 weight percent water and 97.5% wt. pure, and >99.9 area% of (2 r,4 s) -4- { [3, 5-bis (trifluoromethyl) benzyl ] - [5- (3-carboxypropoxy) pyrimidin-2-yl ] amino } -2-ethyl-6-trifluoromethyl-3, 4-dihydro-2H-quinoline-1-carboxylic acid ethyl ester (compound 3).

Example 11.13 additional preparation of Compound 3

Compound 2 was neutralized with aqueous NaOH and dissolved in EtOH. The solution was filtered through activated carbon. Vacuum distillation was performed to concentrate the solution. An aqueous NaOH solution was quantitatively added to obtain a sodium salt of compound 1 in solution, and the ester formed in this and the previous steps was saponified.

Subsequently, a mixture of aqueous CaCl ₂ and EtOAc was prepared in a second vessel. Then the sodium salt of compound 1 from the first vessel was added quantitatively to this mixture, whereby precipitation of compound 3 occurred. Optionally, the suspension may be heated to NMT 25 ℃ and then cooled to 8 ℃. The solid compound 3 was filtered off at 8 ℃, washed with water and dried in vacuo.

EXAMPLE 11.14 grinding of the reprocessed Compound 3

Compound 3 was jet milled using an 8 inch spiral mill. The feed rate, venturi pressure and mill pressure were adjusted within the ranges listed below to enable production of micronized compound 3 meeting the particle size acceptance criteria (d90=6 μm to 15 μm).

The feed rate is 17kg/h to 20kg/h

Grinding pressure 20PSI/1.4 bar

Venturi pressure 100PSI/6.9 bar

Process gas nitrogen

Analysis: mastersizer 3000.

EXAMPLE 11.15 grinding of another preparation of Compound 3

The particle size distribution was adjusted to the target parameters d90:6 to 15 microns by micronization on a screw jet mill, 8 inch jet mill, 8005 and KT4 LIW feeder. The three samples were jet milled with the following results:

d90:8, 8 and 9 microns

D50:4, 3 and 4 microns

D10:2 microns, 1 micron

EXAMPLE 11.16 crystalline calcium obrisedronate

2G of amorphous obiprift-calcium was added to Acetonitrile (ACN)/methyl tert-butyl ketone (MIBK) at a ratio of 6:1 at a concentration of 200mg/ml and the sample was heated to 50 ℃ for 5 minutes until all solids were dissolved. The sample was then placed in a water bath and cooled from 50 ℃ to 5 ℃ at a rate of 0.9 ℃ per minute over 48 hours. The sample was kept at 5 ℃ for 3 days and then transferred to-20 ℃ for 30 minutes before the solids were isolated. The solid was air-dried for 2 hours before further characterization. The process forms crystalline obiprift calcium hemi-hydrate.

EXAMPLE 11.17 Polarized Light Microscopy (PLM)

Polarized light micrographs were taken at room temperature using a Nikon (Nikon) DS-Fi2 upright microscope. Samples (2 mg) were mounted on slides and covered with a drop of silicone oil, with a coverslip on top of the samples for analysis. The samples were not protected from light.

EXAMPLE 11.18-X-ray powder diffraction (XRPD)

XRPD was performed on a silicon zero background mount (silicon zero-background holder) using a PANALYTICAL X' Pert ³ powder diffractometer using an incident beam of Cu radiation generated using a finely focused source epstein tube (Empyran tube). Prior to analysis, a silicon standard (NIST SRM 640 d) was analyzed to verify that the Si 111 peak position was consistent with the NIST certification site. About 5mg to 10mg of the sample was placed on a silicon zero background stand and manually flattened using an aluminum spatula (spatula) to minimize the difference in total height of the sample. The rack was then loaded onto the instrument for analysis. The XRPD parameters used are listed in table 58.

Table 58 parameters for XRPD testing

EXAMPLE 11.19-X-ray powder diffraction pattern

Using PANALYTICAL X-ray powder diffractometer, the measurement conditions were as follows, data were collected by data observer (DataViewer) and data were evaluated by the addition of the exquisiteness fractions (X' Pert High Score Plus):

EXAMPLE 11.20-X-ray powder diffraction pattern

The diffraction pattern of fig. 51 was measured in transmission mode using an Empyrean powder diffractometer from malvern panaceae (MALVERN PANALYTICAL). Samples were prepared as thin layers between two kapton foils and measured in continuous mode. The measuring range of the detector is approximately 2 deg. 2 theta to 40 deg. 2 theta. Peaks are seen at signals at about 3.4 degrees 2 theta, about 7.0 degrees 2 theta, and about 9.2 degrees 2 theta. A peak at about 5.6 ° 2θ is assigned to the kapton foil.

EXAMPLE 11.21X-ray powder diffraction method of crystalline Obiprift HCl/Compound 1D

The diffraction pattern was measured using a siemerzem feier science ARL elkunnox (Thermo FISHER SCIENTIFIC ARL equipenox) 1000 powder diffractometer. The diffractometer is equipped with a copper source and a germanium (111) monochromator providing monochromatic Cu ka 1 radiation, and a position sensitive gas ionization detector.

The samples were measured in reflection mode using an Al sample holder without any further preparation (i.e. grinding). The detector measures the entire angular range of about 2 deg. 2 theta to 120 deg. 2 theta simultaneously, and in the case of HCl obbestrepide, a recognizable signal for phase identification is seen up to about 45 deg. 2 theta. During the measurement, the temperature of the diffractometer is typically around 30 ℃.

EXAMPLE 11.22X-ray powder diffraction method of crystalline Obiprift HCl

The smart laboratory (Rigaku SmartLab) X-ray diffractometer is configured as Bragg-brentgenor reflection geometry (Bragg-Brentano reflection geometry) equipped with beam stop and knife edge (KNIFE EDGE) to reduce incident beam and air scatter. The data collection parameters are shown in table 59.

TABLE 59X-ray powder diffraction parameters

EXAMPLE 11.23 FT-IR Spectroscopy

Fourier transform infrared spectroscopy (FTIR) spectra of samples of amorphous obipratropium calcium are shown in fig. 52. FTIR spectra were obtained using a Bruker Tensor27 spectrometer with Platinum ATR-QL-Diamond units (Platinum ATR-QL-Diamond units). The ground sample was placed on an ATR unit without any pretreatment.

Examples 11.24- ¹ H NMR Spectroscopy

The NMR spectrum of the solution made from the sample of amorphous obiprift halflic calcium is shown in fig. 53. NMR spectra were obtained using 600MHz AVANCE NEO Bruker and in deuterated MeOH as solvent using Tetramethylsilane (TMS) as an internal reference for chemical shift at 0.0 ppm. The spectral shift is consistent with the chemical structure.

EXAMPLE 11.25 modulated differential scanning calorimetry (mDSC)

Samples with the mDSC thermograms shown in fig. 60 and 62 were prepared using a tare (Tzero) aluminum pan with pinholes. The ramp rate is from 25 ℃ to 225 ℃ at a rate of 2 ℃ per minute, with a modulation of ± 0.5 ℃ every 60 seconds. The instrument used was a TA Q2500 DSC from TA Instruments.

EXAMPLE 11.26 modulated differential scanning calorimetry (mDSC)

Using a TA instrument DSC2500, the onset temperature was 25 ℃, and the sample was heated to 225 ℃ at a rate of 2 ℃ per minute per 60 seconds modulation ± 0.5 ℃. For this test Tzero aluminum discs with factory pinholes and Tzero sealing caps were used, which were additionally pierced and enlarged. An integrated thermogram from the sample (showing reverse heat flow) is included in fig. 61. The sample exhibits a glass transition with a Tg of about 111 ℃.

EXAMPLE 11.27 method for assessing stability

Stability studies were performed on crystalline and amorphous forms of obiprifepin at 70 ℃ per 75% Relative Humidity (RH). The solid was placed in 4.0ml uncovered glass bottles (open condition) and stored at 70 ℃ per 75% rh. Samples were pulled from the stabilization chamber at day 1 (24 hours) and day 7 time points. The physical stability of the solid was analyzed by XRPD and the chemical purity was analyzed by HPLC. Samples collected at each time point were dissolved in methanol prior to HPLC analysis. To minimize the effect of potential analyte adsorption onto the filter, the initial 0.5mL supernatant passing through the filter was discarded before the sample was collected for HPLC analysis. The purity of each sample was determined based on the percentage of peak area and compared to the sample at t=0.

EXAMPLE 11.28-method for assessing dynamic solubility in a Biorelevant Medium

Dynamic solubility studies were performed on crystalline and amorphous forms of olanzapine in a biologically relevant medium comprising fed state simulated intestinal fluid (FeSSIF) at pH 5.0 and fasted state simulated intestinal fluid (FaSSIF) at pH 6.5 at 37 ℃. The solids were magnetically stirred in the shaking bath at 600RPM and samples were withdrawn at t=15 minutes, 30 minutes, 60 minutes, 90 minutes and 120 minutes using a 1.0ml syringe. Solubility was measured using the HPLC method provided by the customer. The compound was added to a 4.0ml glass bottle at a concentration of about 20.0 mg/ml. The sample was stirred using a vortex mixer for about 5 minutes to ensure that undissolved excess powder was present. Samples collected at each time point were centrifuged at 1200RPM and filtered using a 0.45 μm Polytetrafluoroethylene (PTFE) filter and diluted with methanol prior to HPLC analysis. To minimize the effect of potential analyte adsorption onto the filter, the initial 0.5mL supernatant passing through the filter was discarded before the sample was collected for HPLC analysis.

While the invention has been particularly shown and described with reference to a preferred embodiment and various alternative embodiments, it will be understood by those skilled in the relevant art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.

List of abbreviations:

API active pharmaceutical ingredient

Degree C

CC compaction compression coefficient (=100deg.X (TBD-IBD)/TBD)

CFM cubic feet per minute

CoA analysis certificate

DBD dynamic total Density (= (TBD-IBD) ²/TBD ] +IBD)

E.g. e.g

EZE/EZE ezetimibe

FDC1 formulation development composition 1

FDC2 formulation development composition 2

G

H Hausner ratio (Hausner ratio) (=TBD/IBD)

HPLC high performance liquid chromatography

IBD initial total density (=mass/initial volume)

KF Karl Fischer

Kp kilopounds

KN kilonewton

LC tag declaration

LLS laser light scattering

Lt liter of

Mg

Min

ML of

Mm millimeter

N.R is not recorded

N.D does not detect

Obi/OBI Obipsilatep (also called TA-8995)

PH-log [ H ⁺ ] or pH= -log a _H ⁺

PSD particle size distribution

RH relative humidity (a _w. Times.100)

Rpm/min

RRT relative retention time

RSD relative standard deviation

Sec seconds

Standard deviation of SD

SLS Kolliphor SLS fines, EP & USP/NF & JP

T temperature (° C)

TBD tap bulk Density (=Mass/tap volume)

Vs relative to

W/w weight/weight

XRPD X-ray powder diffraction

Micron μmDiameter of

% A/a impurity area is the total area of peaks not present in the blank

Claims

1. A fixed-dose pharmaceutical composition, comprising or consisting of:

a. orbicetrapib or a pharmaceutically acceptable salt, solvate or cocrystal thereof,

b. ezetimibe or a pharmaceutically acceptable salt, solvate or cocrystal thereof, and

c. one or more pharmaceutically acceptable excipients.

2. The pharmaceutical composition of claim 1, wherein when the pharmaceutical composition is orally administered to a subject, the 90% confidence interval of the geometric mean of the area under the curve (AUC _0-∞ and/or AUC _0- t ) and/or Cmax of orbicetrapib is within the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t ) and/or Cmax of orbicetrapib obtained when a reference pharmaceutical composition is orally administered to similar subjects, respectively, wherein the reference pharmaceutical composition comprises an equivalent dose of orbicetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone, or co-administered simultaneously or sequentially with another pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, or administered in the form of a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

3. The pharmaceutical composition of any of the preceding claims, wherein when the pharmaceutical composition is orally administered to a subject, the 90% confidence interval for the geometric mean of the area under the curve (AUC _0-∞ and/or AUC _{0-t ) and/or Cmax of ezetimibe and/or ezetimibe glucuronide, respectively, is approximately 0.01% of the area under the curve (AUC 0-} _∞ and/or AUC _0-t ) of ezetimibe and/or ezetimibe glucuronide, respectively, obtained when a reference pharmaceutical composition is orally administered to a similar subject. ) and/or Cmax is within the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110%, wherein the reference pharmaceutical composition comprises an equivalent dose of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone, or co-administered simultaneously or sequentially with another pharmaceutical composition comprising orbicetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, or administered in the form of a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

4. A pharmaceutical composition according to any one of the preceding claims, for use in treating a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, wherein the pharmaceutical composition is deemed suitable for said use in the following circumstances:

a. Orally administering the fixed dose pharmaceutical composition to a subject;

b. measuring the concentration of orbitrap in the subject's plasma at one or more time points after administration to provide a set of orbitrap concentration/time data points to provide an area under the curve (AUC); and

c. the 90% confidence interval for the geometric mean of the area under the curve (AUC _0-∞ and/or AUC 0- _t ) and/or Cmax of orbicetrapib is within the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t ) and/or Cmax of orbicetrapib, respectively, obtained when the reference pharmaceutical composition is orally administered to similar subjects, wherein the reference pharmaceutical composition comprises an equivalent dose of orbicetrapib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone, or co-administered simultaneously or sequentially with another pharmaceutical composition comprising ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, or administered in the form of a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

5. A pharmaceutical composition according to any one of the preceding claims, for use in treating a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, wherein the pharmaceutical composition is deemed suitable for said use in the following circumstances:

a. Orally administering the fixed dose pharmaceutical composition to a subject;

b. measuring the concentration of ezetimibe and/or ezetimibe glucuronide in the subject's plasma at one or more time points after administration to provide a set of ezetimibe and/or ezetimibe glucuronide concentration/time data points, respectively, thereby providing an area under the curve (AUC) for ezetimibe and/or ezetimibe glucuronide, respectively; and

c. the 90% confidence interval for the geometric means of the area under the curve (AUC _0-∞ and/or AUC _0-t ) and/or Cmax for ezetimibe and/or ezetimibe glucuronide is within the range of 75% to 125%, preferably 80% to 125%, and more preferably 90% to 110% of the area under the curve (AUC _0-∞ and/or AUC _0-t ) and/or Cmax for ezetimibe and/or ezetimibe glucuronide, respectively, obtained when the reference pharmaceutical composition is orally administered to similar subjects, wherein the reference pharmaceutical composition comprises an equivalent dose of ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and wherein the reference pharmaceutical composition is administered alone or co-administered simultaneously or sequentially with another pharmaceutical composition comprising orbicetamipib or a pharmaceutically acceptable salt, solvate or co-crystal thereof, or administered in the form of a fixed dose combination with ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof.

6. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the use of the pharmaceutical composition is for lowering LDL cholesterol and/or increasing HDL cholesterol in a person suffering from heterozygous familial hypercholesterolemia (HeFH) and/or diagnosed with atherosclerotic cardiovascular disease (ASCVD).

7. The pharmaceutical composition of any of the preceding claims, wherein t of AUC 0-t is selected from 48 hours (AUC 0-48), 72 hours (AUC 0-72), 96 hours (AUC 0-96), 144 hours (AUC 0-144), 192 hours (AUC 0-192), 240 hours (AUC 0-240), 336 hours (AUC 0-336) or AUC 0-∞, preferably 48 hours (AUC 0-48), and more preferably AUC 0-∞.

8. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the subject is a human, preferably a healthy human, more preferably a human in need of lowering LDL cholesterol and/or increasing HDL cholesterol, a human suffering from heterozygous familial hypercholesterolemia (HeFH) and/or a human diagnosed with atherosclerotic cardiovascular disease (ASCVD).

9. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the subject is a healthy adult man or woman aged 18 to 65 years who does not use tobacco or nicotine, and optionally, the human has a body mass index of 18.5 to 29.9 Kg/ ^m2 .

10. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the LDL cholesterol level of the person in need of lowering LDL cholesterol, and/or the person suffering from heterozygous familial hypercholesterolemia (HeFH), and/or the person diagnosed with atherosclerotic cardiovascular disease (ASCVD) is ≥70 mg/dL, and optionally, the person is not adequately controlled by their current lipid-lowering therapy.

11. A pharmaceutical composition as claimed in any of the preceding claims, wherein at least about 60%, preferably at least about 70%, and more preferably at least about 80% of the ezetimibe is dissolved in about 30 minutes when the pharmaceutical composition is dissolved in 500 ml of a solution comprising 0.45% SLS in 0.05 M sodium acetate buffer, pH 4.5, at about 75 rpm in a USP II type apparatus.

12. The pharmaceutical composition of any of the preceding claims, wherein at least about 70%, preferably at least about 80%, and more preferably at least about 85% of orbicetrapib is dissolved in about 15 minutes when the pharmaceutical composition is dissolved in 1000 ml of a solution comprising a phosphate buffer solution at pH 6.8 + 0.2% w/v polysorbate 80 in a USPI type II apparatus at about 75 rpm at 37±0.5°C.

13. The pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutical composition comprises 1 mg to 20 mg of orbicetrapib and 5 mg to 20 mg of ezetimibe, preferably the pharmaceutical composition comprises 5 mg of orbicetrapib and 10 mg of ezetimibe or 10 mg of orbicetrapib and 10 mg of ezetimibe.

14. The pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutical composition is provided in a unit dosage form comprising 1 mg to 20 mg of orbicetrapib and 5 mg to 20 mg of ezetimibe, preferably the unit dosage form comprises 5 mg of orbicetrapib and 10 mg of ezetimibe, or 10 mg of orbicetrapib and 10 mg of ezetimibe.

15. The pharmaceutical composition of any of the preceding claims, wherein the pharmaceutical composition comprises no more than about 2% (w/w), preferably no more than about 0.5% (w/w), more preferably no more than about 0.3% (w/w), even more preferably no more than about 0.2% (w/w) of an ezetimibe tetrahydropyran analog as an impurity.

16. The pharmaceutical composition of any one of the preceding claims, wherein ezetimibe or orbicetrapib, or both ezetimibe and orbicetrapib, are micronized.

17. A pharmaceutical composition according to any one of the preceding claims, wherein the pharmaceutical composition comprises micronized ezetimibe having a Dv90 of no more than 10 μm, preferably in the range of 4 to 10 μm, more preferably no more than 8.5 μm; _{a Dv50} of no more than 4 μm, preferably in the range of about 1 to 4 μm, more preferably no more than 3.8 μm; and _a _Dv10 of no more than 1 μm.

18. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition comprises micronized orbicetrapib having a Dv90 of no more than 14 μm, preferably in the range of about 5 μm to 14 μm; a _Dv50 of no more than 5 μm, preferably in the range of about 3 μm to 5 μm; and a _Dv10 of no more than ₃ μm.

19. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition comprises ezetimibe in the form of anhydrous ezetimibe, ezetimibe monohydrate, or a mixture thereof.

20. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition comprises orbicetrapib in the form of an alkali metal or alkaline earth metal salt of orbicetrapib, preferably orbicetrapib sodium, orbicetrapib potassium or orbicetrapib calcium, and more preferably orbicetrapib calcium salt.

21. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition is a two-component composition, and wherein one component of the two-component composition comprises ezetimibe and the other component comprises orbicetrapib.

22. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition is a two-component composition, and wherein one component of the two-component composition comprises both ezetimibe and orbicetrapib.

23. The pharmaceutical composition of any one of claims 21 to 22, wherein the two-component composition is a bilayer tablet formulation, a capsule formulation comprising or consisting of two types of granules, or a tablet formulation comprising an extragranular component and an intragranular component.

24. The pharmaceutical composition of claim 22, wherein:

a. the intragranular component comprises ezetimibe, and the extragranular component comprises orbicetrapib; or

The intragranular components include both ezetimibe and orbicetrapib; and the extragranular components include only excipients.

25. The pharmaceutical composition of claim 22, wherein:

a. the intragranular component comprises orbicetrapib, and the extragranular component comprises ezetimibe; or

b. The extragranular components include both ezetimibe and orbicetrapib, and the intragranular components include only excipients.

26. A pharmaceutical composition as claimed in any of the preceding claims, wherein the pharmaceutical composition further comprises one or more binders and surfactants, wherein preferably the ratio of binder:surfactant in the intragranular component is in the range of about 0.05:5.0 to about 5.0:0.05, preferably about 0.5:4.5 to about 4.5:0.5, more preferably about 1:4 to about 4:1, even more preferably about 1:2 to about 2:1, and most preferably about 1:1.

27. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the pharmaceutical composition further comprises one or more binders selected from the group consisting of cellulose derivatives, preferably selected from methylcellulose and carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxypropylcellulose and hydroxyethylcellulose; gelatin, glucose, dextrose, xylitol, polymethacrylates, polyvinylpyrrolidone and copolymers of polyvinylpyrrolidone, starch paste, sucrose, sorbitol, pregelatinized starch, tragacanth gum, alginic acid and salts of alginic acid such as sodium alginate, magnesium aluminum silicate, polyethylene glycol, guar gum, bentonite, preferably the binder is polyvinylpyrrolidone or a copolymer of polyvinylpyrrolidone, more preferably the binder is copovidone, and even more preferably the binder is Colidan 30.

28. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the pharmaceutical composition further comprises one or more surfactants having an HLB value of at least 15, at least 20, at least 30 or at least 40; preferably, the one or more surfactants are selected from lauric acid or a salt of lauric acid, palmitic acid or a salt of palmitic acid, stearic acid or a salt of stearic acid and oleic acid or a salt of oleic acid, polyethylene glycol glycerides, polyoxyethylene monoesters, polyoxyethylene ethylene monostearate, polyoxyethylene monolaurate, polyoxyethylene sorbitan monooleate, polyethoxylated castor oil, polyethylene glycol having a molecular weight in the range of about 2000 to 10000, propylene glycol caprylate, glyceryl oleate and caprylate, esters of glycerol and fatty acids; more preferably, the one or more surfactants are selected from dioctyl sodium sulfosuccinate, Capmul PG-8, Capryol 90, Capmul MCM, polysorbate 20, polysorbate 40 or polysorbate 80 or sodium lauryl sulfate; and even more preferably, the surfactant is sodium lauryl sulfate.

29. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition further comprises one or more disintegrants selected from cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, alginic acid, sodium alginate, microcrystalline cellulose, sodium starch glycolate, pregelatinized starch, preferably the disintegrant is cross-linked sodium carboxymethyl cellulose or sodium starch glycolate, and more preferably the disintegrant is sodium starch glycolate.

30. A pharmaceutical composition as claimed in any of the preceding claims, wherein the pharmaceutical composition remains stable at 40°C/75% relative humidity for at least 1 month, preferably at least 3 months and more preferably at least 6 months, or at 25°C/60% relative humidity for at least 3 months, preferably at least 6 months and more preferably at least 12 months.

31. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the pharmaceutical composition is a tablet formulation comprising or consisting of:

a. Intragranular components, comprising:

i. orbicetrapib calcium equivalent to 10 mg orbicetrapib free acid;

ii. anhydrous ezetimibe or a mixture of anhydrous ezetimibe and ezetimibe hydrate equivalent to 10 mg ezetimibe;

iii. A binder to a surfactant in a ratio of 1:1, preferably the binder and the surfactant each account for about 1±0.5% w/w of the granules of the intragranular component; more preferably, the binder is 1±0.5% w/w of povidone or polyvinylpyrrolidone, and the surfactant is 1±0.5% w/w of sodium lauryl sulfate;

iv. a disintegrant selected from cross-linked sodium carboxymethylcellulose, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate; preferably the disintegrant accounts for about 2% w/w to 8% w/w of the granule of the intragranular component, preferably 3% w/w to 6% w/w, more preferably about 4.5±0.5% w/w;

v. one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably anhydrous lactose or lactose monohydrate, even more preferably lactose monohydrate; polysaccharides, preferably cellulose, more preferably microcrystalline cellulose; sugar alcohols, preferably sorbitol, xylitol or mannitol;

b. Extragranular components, comprising:

i. a disintegrant selected from microcrystalline cellulose, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate;

ii. optionally a lubricant, preferably magnesium stearate,

iii. optionally a glidant, preferably colloidal silicon dioxide or talc or both;

iv. optionally one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably anhydrous lactose or lactose monohydrate, even more preferably lactose monohydrate; polysaccharides, preferably cellulose, more preferably microcrystalline cellulose; sugar alcohols, preferably sorbitol, xylitol or mannitol; more preferably mannitol and microcrystalline cellulose;

c. Optionally, the pharmaceutical composition comprises a film coating, preferably the film coating does not contain primary alcohol, more preferably the film coating does not contain polyethylene glycol.

32. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the pharmaceutical composition comprises a tablet formulation comprising or consisting of:

a. Intragranular components, comprising:

i. anhydrous ezetimibe or a mixture of anhydrous ezetimibe and ezetimibe hydrate equivalent to 10 mg ezetimibe;

ii. A binder to a surfactant in a ratio of 1:1, preferably the binder and the surfactant each account for about 1±0.5% w/w of the granules of the intragranular component; more preferably, the binder is 1±0.5% w/w of povidone or polyvinylpyrrolidone, and the surfactant is 1±0.5% w/w of sodium lauryl sulfate;

iii. a disintegrant selected from cross-linked sodium carboxymethylcellulose, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate; preferably the disintegrant accounts for about 2% w/w to 8% w/w of the granule of the intragranular component, preferably 3% w/w to 6% w/w, more preferably about 4.5±0.5% w/w;

iv. one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably anhydrous lactose or lactose monohydrate, even more preferably lactose monohydrate; polysaccharides, preferably cellulose, more preferably microcrystalline cellulose; sugar alcohols, preferably sorbitol, xylitol or mannitol;

b. Extragranular components, comprising:

i. orbicetrapib calcium equivalent to 10 mg orbicetrapib free acid;

ii. a disintegrant selected from cross-linked sodium carboxymethyl cellulose, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate;

iii. optionally a lubricant, preferably magnesium stearate,

iv. optionally a glidant, preferably colloidal silicon dioxide or talc or both;

v. optionally one or more diluents selected from disaccharides, preferably lactose or sucrose, more preferably anhydrous lactose or lactose monohydrate, even more preferably lactose monohydrate; polysaccharides, preferably cellulose, more preferably microcrystalline cellulose; sugar alcohols, preferably sorbitol, xylitol or mannitol; more preferably mannitol and microcrystalline cellulose;

33. A pharmaceutical composition as claimed in any one of the preceding claims, wherein the pharmaceutical composition comprises a tablet formulation comprising or consisting of:

a. Intragranular components, comprising:

i. orbicetrapib calcium equivalent to 10 mg orbicetrapib free acid;

b. Extragranular components, comprising:

ii. a disintegrant selected from microcrystalline cellulose, pregelatinized starch or sodium starch glycolate, more preferably sodium starch glycolate;

iii. optionally a lubricant, preferably magnesium stearate,

iv. optionally a glidant, preferably colloidal silicon dioxide or talc or both;

34. A pharmaceutical composition according to any one of the preceding claims for use in the treatment of a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, preferably a subject suffering from hyperlipidemia or mixed dyslipidemia.

35. A pharmaceutical composition as claimed in any of the preceding claims, for use in lowering LDL cholesterol in patients in need of lowering LDL cholesterol and/or raising HDL cholesterol, patients with heterozygous familial hypercholesterolemia (HeFH) and/or patients diagnosed with atherosclerotic cardiovascular disease (ASCVD).

36. Use of a pharmaceutical composition as claimed in any one of the preceding claims for the preparation of a medicament for treating a subject suffering from hyperlipidemia or mixed dyslipidemia.

37. Use of a pharmaceutical composition as claimed in any one of the preceding claims in the preparation of a medicament for lowering LDL cholesterol in a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, a subject with heterozygous familial hypercholesterolemia (HeFH) and/or a subject diagnosed with atherosclerotic cardiovascular disease (ASCVD).

38. Use of a pharmaceutical composition as claimed in any one of the preceding claims in the preparation of a medicament for reducing the risk of cardiovascular events.

39. The use of any one of the preceding claims, wherein the subject suffers from mild dyslipidemia.

40. A method for treating a subject in need of lowering LDL cholesterol and/or raising HDL cholesterol, a subject with heterozygous familial hypercholesterolemia (HeFH), and/or a subject diagnosed with atherosclerotic cardiovascular disease (ASCVD), wherein the method comprises administering to a patient in need thereof a therapeutically effective dose of a pharmaceutical composition as claimed in any one of the preceding claims.

41. A method of treating a subject suffering from hyperlipidemia or mixed dyslipidemia, wherein the method comprises administering to a patient in need thereof a pharmaceutical composition as claimed in any one of the preceding claims.

42. The use of a pharmaceutical composition as claimed in any one of the preceding claims in the preparation of a medicament or a method of treatment, wherein the subject's LDL-cholesterol level is ≥50 mg/dL, preferably ≥70 mg/dL, and optionally, the subject is not adequately controlled by their current lipid-lowering therapy.

43. Use of a pharmaceutical composition as claimed in any of the preceding claims, wherein the pharmaceutical composition is administered to the subject in need thereof to deliver a total daily oral dose of 5 mg orbicetrapib and 10 mg ezetimibe, 10 mg orbicetrapib and 10 mg ezetimibe, or 20 mg orbicetrapib and 20 mg ezetimibe, preferably the pharmaceutical composition is administered to the subject to deliver a daily oral dose of 10 mg orbicetrapib and 10 mg ezetimibe.

44. Use of a pharmaceutical composition as claimed in any of the preceding claims, wherein the subject in need is a subject who needs to additionally lower low-density lipoprotein cholesterol as an adjunct to diet and/or maximally tolerated lipid-lowering therapy for the treatment of adults with heterozygous familial hypercholesterolemia (HeFH) or diagnosed with atherosclerotic cardiovascular (CV) disease (ASCVD).

45. A pharmaceutical composition comprising orbicetrapib and ezetimibe or a pharmaceutically acceptable salt, solvate or co-crystal thereof, and a pharmaceutically acceptable carrier, for use in treating a subject who requires additional lowering of low-density lipoprotein cholesterol as an adjunct to diet and/or maximally tolerated lipid-lowering therapy for treating adults with heterozygous familial hypercholesterolemia (HeFH) or diagnosed with atherosclerotic cardiovascular (CV) disease (ASCVD).