HK1170631B - Novel compositions and processes for preparing 5-amino or substituted amino 1,2,3-triazoles and triazole orotate formulations - Google Patents

Description

Novel compositions and methods for preparing 5-amino or substituted amino 1,2, 3-triazoles and triazole orotate formulations

Cross reference to other applications

This application is a continuation-in-part application of U.S. patent application 12/584,448 filed on 9, 4, 2009, which is incorporated herein by reference in its entirety.

1. Field of the invention

The present invention relates to novel compounds of 5-amino or substituted amino 1,2, 3-triazoles and substituted derivatives thereof (herein referred to as Carboxyamidotriazoles (CAIs) or CAIs), to formulations of 5-amino or substituted amino 1,2, 3-triazoles orotate and substituted derivatives thereof, (having a specific base structure (base): acid ratio, CTOs), to formulations of 5-amino or substituted amino 1,2, 3-triazoles orotate and substituted derivatives thereof and orotic acid, (having a specific base structure: acid ratio, CAOs), and to a safer process for preparing the same, the method synthesizes intermediate azide materials by using stable, more efficient and safer raw materials, the azide material is essential in the synthetic pathway of CAI, and orotate agents CTO and CAO. More particularly, the present invention relates to novel polymorphs (polymorphs) of 5-amino or substituted amino 1,2, 3-triazoles and substituted derivatives thereof. Even more particularly, the present invention relates to novel 5-amino or substituted amino 1,2, 3-triazole orotate (CTOs having optimal base structure: acid ratio ranging from 1:1 to 1: 4), and formulations of 5-amino or substituted amino 1,2, 3-triazole and substituted derivatives thereof and orotic acid (CAO) in optimal base structure: acid ratio ranging from 1:1 to 1:4, and to the use of the same for the control and treatment of diseases including but not limited to solid cancer, macular degeneration, retinopathy, chronic myeloid leukemia, AIDS and diseases that rely on aberrant signal transduction and proliferation.

2.Background

The present invention is in the field of developing new polymorphs of 5-amino or substituted amino 1,2, 3-triazole (CAI) and its substituted derivatives, developing orotate salts of 5-amino or substituted amino 1,2, 3-triazole and its substituted derivatives, and developing formulations of 5-amino or substituted amino 1,2, 3-triazole and its substituted derivatives and orotic acid (in optimal ratio of basic structure: acid). The present invention aims to develop new polymorphs of 5-amino or substituted amino 1,2, 3-triazole and substituted derivatives thereof to improve chemical, biological, pharmacokinetic and toxicokinetic properties and to improve therapeutic properties, including but not limited to: anti-cancer activity, anti-metastatic activity, calcium-mediated signal transduction, anti-angiogenesis, anti-PI 3, anti-COX 2, apoptosis, down-regulation of BCR-ABL protein in chronic myeloid leukemia, regulation of HIVLTR transcription, or anti-VEGF 1 properties.

In 1986, 5-amino or substituted amino 1,2, 3-triazole compounds and their substituted derivatives were demonstrated to have anticoccidial activity. U.S. Pat. No.4,590,201 (issued to R.J. Bochis et al, 1986) describes a process for the preparation of 5-amino-1- (4- [ 4-chlorobenzoyl ] -3, 5-dichlorobenzyl) -1,2, 3-triazole-4 carboxamide (L651582 or CAI) which involves the use of 3, 5-dichloro-4- (4-chlorobenzoyl) benzyl azide, an important intermediate in the sodium azide synthetic pathway. Subsequently, it was demonstrated that L651582 or CAI inhibits selected signal transduction pathways, including those involving calcium influx, release of arachidonic acid, and production of inositol phosphates. U.S. patent 5,359,078 (issued to e.c. kohn et al, 1994). "L651582" as used herein denotes L6515182, CAI, carboxyamidotriazole, NSC609974 or 99519-84-3 as described in the prior art.

U.S. patent No.5,912,346 (issued to f. wehrmann,1999) subsequently discloses inorganic and organic salts of L651582, and specifically describes a process for the preparation of the orotate salt of L651582. L651582 was prepared by the method described in us patent No.4,590,201. The L651582: orotate ratio was confirmed by proton NMR to be 2:1 (base structure: acid) and to have a melting point of 234 ℃ and 235 ℃. The synthesis of intermediate 3- (4-chlorobenzoyl) -4-chlorobenzyl azide was performed as described above using intermediate 3- (4-chlorobenzoyl) -4-chlorobenzoyl bromide in ethanol and sodium azide. U.S. Pat. No.5,912,346 describes improved antitumor activity of L651582 orotate (CAI orotate, basic structure: acid, 2:1) compared to the equivalent dose of L651582 in the rat androgen-independent Dunning R-3227-AT-1 prostate cancer model.

Carboxyaminotriazole, L651582, CAI, NSC609974, or 99519-84-3, as an inhibitor of calcium-mediated signal transduction, was the first to be discovered as a cell proliferation inhibitory signal-inhibiting anticancer drug. The above drugs were tested in patients with stage I, II, and III solid cancers at the national cancer institute (national cancer institute). However, NCI has terminated the development of L651582 because it has not proven effective in human trials and/or suffers from poor bioavailability, severe gastrointestinal toxicity, neurotoxicity, and tolerability issues have prevented the use of optimal dosages to achieve therapeutic effects. Capsules of micronized formulation of L651582 in PEG-400 were used in clinical studies to improve the bioavailability of the drug. KohneC et al, clinical cancer Res7: 1600-; BauerKS et al, clinical cancer Res5:2324-2329 (1999); BerlinJ et al, JClinOnc15:781-789 (1997); BerlinJ et al, clinical cancer 8:86-94 (2002); YasuiH et al, JBiolChem45:28762-28770 (1997); alessandro et al, JCellPhysiol215: 111-.

Thus, L651582 orotate (basic structure: acid 2:1), described in U.S. Pat. No.5,912,346, represents a potential way to reuse this promising drug, improving the efficiency of L651582 based on preclinical studies. However, according to the process described in U.S. Pat. No.5,912,346, problems were encountered in scaling up the process for preparing L651582 orotate in large quantities (2:1 ratio).

In relation to the use of orotic acid to potentiate the analgesic effect of drugs, U.S. Pat. No.4,061,741 (issued to WawretschekW et al, 1977) describes the use of a combination of dexpropoxyphene-HCl, levopropoxyphene (laevopropyphene) -HCl, or sodium salicylate and choline orotate, showing that pharmaceutical formulations combined with choline orotate provide the best results. It is clear that the prior art shows a reverse teaching regarding the ratio and chemical nature of the binding of orotic acid to the compound.

The synthesis scheme described in the prior art for the L651582 orotate salt is shown in reaction scheme I above. 858 is a product identifier (identifier) and, for example, 858A-858D represents an intermediate. 858E represents Carboxyamidotriazole (CAI). 858F represents carboxyamidotriazole orotic acid or carboxyamidotriazole orotate or CTO as used herein.

The prior art teachings show that the combined use of choline orotate and a pharmaceutical is a preferred embodiment. Unfortunately, however, such methods do not address the problems encountered in the present scale-up process for the preparation of CTO for clinical development. It is not clear that the basic structure to acid ratio (2:1) in L651582 orotate is the optimal chemical structure for the drug. In addition, problems were encountered in scaling up the preparation of L651582 orotate (2:1) for mass production. Few manufacturers have equipment and devices that can handle large amounts of sodium azide, and those contractors with the devices charge a large amount of service.

After the step of protecting the alcohol group in 3, 5-dichlorobenzyl alcohol, i.e., TBDMS ether (step 1), the ether is reacted with 4-chlorobenzoyl chloride to form a substituted benzophenone (step 2). The benzophenone is treated with thionyl chloride (step 3) followed by sodium azide (step 4) to form 3, 5-dichloro-4- (4-chlorobenzoyl) benzylazide. Reaction of the azide with cyanoacetamide yielded L651582 (step 5). Reaction of L651582 with orotic acid formed L651582 orotate (2:1) (step 6).

The use of sodium azide in step 4 of the above process is a serious obstacle in scaling up the production of L6515182 orotate for mass production. Handling large amounts of sodium azide requires carrying out in dedicated pressure sensitive reactors, since sodium azide is a high internal energy (highernergycontent) toxic substance. Such dedicated containment devices for handling sodium azide generally increase the cost of manufacture because fewer pharmaceutical manufacturers have the ability to scale up the production of large quantities of the drug. This is because sodium azide is a fast-reacting, potentially lethal chemical constituent in the form of an odorless white solid. When mixed with water or acid, sodium azide rapidly changes to a toxic gas with an irritating odor. Sodium azide also changes to a toxic gas when it comes into contact with solid metals. Survivors of severe sodium azide poisoning may have cardiac and brain damage, and the centers for disease prevention control (center) advises victims of immediate hot-line contact. (CDC-an issue with sodium azide (FactsAbout SodiumAzide), 2009. apparently, there is a need to develop a safer, new, affordable, and efficient method for preparing L651582 orotate without the use of sodium azide.A competitive bid on affordable costs is not possible because, as shown above, sodium azide is required in the preparation of 3, 5-dichloro-4- (4-chlorobenzoyl) benzyl azide (which is an intermediate in the L651582 orotate synthesis pathway). therefore, there is a need to develop an alternative, safer and more efficient method for preparing orotate drugs with optimal chemical configuration and basic structure: acid ratio.

Even though L651582 orotate has been demonstrated to have significantly higher antitumor activity in rat models of prostate cancer (U.S. patent No.5,912,346), there is no teaching or suggestion as to whether the chemical, pharmacological and biological properties of L651582 orotate are optimal in the 2:1 ratio of basic structure to acid. Clearly, there is a need to develop new polymorphic forms of CAI and orotate compounds of CAI that provide optimal chemical, biological, pharmacological, therapeutic and toxicokinetics to prove useful for clinical development.

It is therefore a main object of the present invention to develop an orotate formulation of CAI (where the ratio of basic structure: acid ranges from 1:1 to 1: 4) with improved efficacy related to bioavailability, which is dependent on its solubility in human body fluids.

It is a further object of the present invention to develop a safer, more cost-effective method for the mass production of CAI, CTO (orotate salt of CAI) and CAO (formulation of CAI mixed with orotic acid).

An important object of the present invention is to produce safer CAI using safer and less toxic ingredients instead of using sodium or potassium azide (which is highly toxic at very low concentrations) to produce intermediates. CAI prepared by the methods described in the prior art has been found to cause severe neurotoxicity and stomach toxicity in patients. It is therefore important to use safer ingredients and to use an improved process for the production of new polymorphic forms of CAI and its orotate formulations.

The relevant subject matter of the above references is specifically incorporated herein by reference in its entirety.

3. Summary of the invention

The present invention seeks to overcome the disadvantages inherent in the prior art by providing compositions of new polymorphs of 5-amino or substituted amino 1,2, 3-triazole and its substituted derivatives (referred to herein as carboxyamidotriazole or CAI), formulations of 5-amino or substituted amino 1,2, 3-triazole orotate and its substituted derivatives, (with specific base structures: acid ratios, CTOs); and 5-amino or substituted amino 1,2, 3-triazole orotate and its substituted derivatives and preparations of orotic acid, (with specific basic structure: acid ratio, CAOs).

The present invention provides a safer process for the preparation of the above by using stable, more efficient and safer starting materials for the synthesis of intermediate azide materials, which are essential in the synthetic pathways of CAI and orotate formulations CTO and CAO.

More particularly, the present invention relates to novel polymorphs of 5-amino or substituted amino 1,2, 3-triazole (CAI) and substituted derivatives thereof. CAI exists in several polymorphic forms (polymorphicheomorphs) including, but not limited to, Form 1(Form1) or Form 2(Form 2).

Further, the present invention more specifically relates to novel 5-amino or substituted amino 1,2, 3-triazole orotate salts (CTOs having optimal base structure: acid ratio ranging from 1:1 to 1: 4), and preparations of 5-amino or substituted amino 1,2, 3-triazole and substituted derivatives thereof and orotic acid (CAO), Orotate Salts (CTOs) and substituted derivatives thereof in optimal base structure: acid ratio.

In a further aspect, the present invention relates to a process for the preparation of the necessary intermediate azide materials in the synthetic route by using stable, safer and affordable raw materials instead of sodium azide or potassium azide, including but not limited to diphenylphosphoryl azide or trimethylsilyl azide, TMSN 3.

More particularly, the present invention relates to novel polymorphs of 5-amino or substituted amino 1,2, 3-triazoles and their substituted derivatives, their orotate derivatives (CTO) (basic structure: acid ratio in the range of 1:1 to 1: 4), and the use thereof to treat diseases including, but not limited to: solid cancers, macular degeneration, retinopathy, chronic myeloid leukemia, AIDS, and diseases that rely on aberrant signaling and proliferation pathways (such as stress-independent calcium channel blockers, PI3, COX2, BCR-ABL, apoptosis, HIVLTR transcription, or VEGF 1).

In view of the foregoing state of the art, the present invention provides novel orotate derivatives of 5-amino or substituted amino 1,2, 3-triazole or carboxyamidotriazole orotate (CTO) wherein said substances contain chemical organic moieties that increase their bioavailability, increase delivery to targets, increase antitumor efficacy and reduce toxicity. Specifically, carboxyamidotriazole orotate (CTOs) of a class having ionic bonds in a ratio ranging from about 1:1 to 1:4 (triazole: orotic acid) constitute the novel compounds of the present invention.

In addition, formulations of 5-amino or substituted amino 1,2, 3-triazoles (CAI) and substituted derivatives thereof and orotic acid (with specific base structure: acid ratio, 1: 1-1: 4, CAOs).

In a further aspect, the present invention provides for the use of diphenyl phosphorazidate (DPPA) or TMN instead of sodium azide₃Or a safer azide equivalent, to the azide intermediate 3, 5-dichloro-4- (4-chlorobenzoyl) benzyl azide. DPPA is significantly safer than sodium azide and has been used to convert alcohols directly to azides, thus, the step in the synthetic pathway of CTO (step 3 of the above-listed scheme) was omitted.

It is a further object of the invention to improve the bioavailability of CTO by oral or other routes in humans and other mammals and to improve the delivery of CTO to targets, e.g. by improving absorption, transport and transport across tissues, blood brain barrier and chorioretinal complex.

In addition, it is an additional object of the present invention to reduce the toxicity of CTO and related compounds by enhancing the clearance of the drug from blood, tissues and organs when administered in the form of orotate salts.

The present invention can be further used to reduce drug interactions and side effects when CTO or CAI and orotic acid (CAO) are administered in combination as a formulation.

It is a further object of the present invention to provide compositions of CTO for the treatment of human neoplasms (humanneoplasts) and in particular primary or metastatic tumors, diseases involving neovascularization such as macular degeneration, retinopathy, diabetic retinopathy, chronic myeloid leukemia, AIDS and diseases that rely on aberrant signaling and proliferation pathways such as stress-independent calcium channel blockers, PI3, COX2, BCR-ABL, apoptosis, HIVLTR transcription or VEGF1, and to reduce the toxic side effects of drugs by reducing the level of drugs in noncancerous tissues that are targets susceptible to drug toxicity by 10% to 100% compared to L651582 or CAI.

Preferred embodiments of the present invention include a base structure to CTO in a 1:1 acid ratio, more preferred embodiments in a 1:2 ratio, and most preferred embodiments of the present invention include CTO compositions (in a ratio of about 0.7:1.3) prepared by the novel process of the present invention for use in the treatment of diseases including, but not limited to: solid cancers, macular degeneration, retinopathy, chronic myeloid leukemia, and modulation of signal transduction pathways such as PI3, COX2, BCR-ABL, STATS, CrkL, apoptosis, HIVLTR transcription, VEGF1, or others.

4. Brief description of the drawings

Fig. 1 shows by NMR the structure of CTO, in the form of CAI: orotic acid or CAI: orotate, of CTO sample J02642 having form1 or form1 polymorph of CAI.

Fig. 2 shows by NMR the structure of CTO, in the form of CAI: orotic acid or CAI: orotate, of CTO sample J02643 having form2 or form2 polymorph of CAI.

Fig. 3 shows a high resolution diffractogram of CTO sample J02642 having the CAI form1 or form1 polymorph.

Fig. 4 shows a high resolution diffractogram of CTO sample J02643 having the CAI form2 or form2 polymorph.

Fig. 5 shows FT-IR of CTO sample J02642 having CAI form1 or form1 polymorph.

Fig. 6 shows FT-IR of CTO sample J02643 having CAI form2 or form2 polymorph.

5. Detailed description of the invention

The present invention provides novel polymorphs of 5-amino or substituted amino 1,2, 3-triazoles or their substituted amino 1,2, 3-triazoles (CAIs) of the class of compounds of formula I, prepared by a novel process. Novel polymorphs of CAI include, but are not limited to: form1 or form2 as confirmed by techniques such as NMR, DSC, FT-IR and XRDP.

Formula I

Wherein R is₁Has the formula (II) in which,

R₁is that

Wherein p is 0 to 2; m is 0 to 4; and n is 0 to 5; x is O, S, SO₂、CO、CHCN、CH₂Or C = NR₆Wherein R is₆Is hydrogen, lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino or cyano; and R₄And R₅Independently a halogen atom (F, Br, Cl), cyano, trifluoromethyl, lower alkanoyl, nitro, lower alkyl, lower alkoxy, carboxy, lower alkoxycarbonyl, trifluoromethoxy, acetylamino, lower alkylthio, lower alkylsulfinyl, lower alkylsulfonyl, trichloroethyl, trifluoromethylthio, trifluoromethylsulfinyl, or trifluoromethylsulfonyl; r₂Is amino, mono-or di-lower alkylamino, acetamido, acetylimino, ureido, carboxamido, iminomethyl or guanidino; and R₃Is carbamoyl, cyano, carbazolyl, amidino or N-hydroxycarbamoyl; wherein said lower alkyl group containsThe lower alkyl, lower alkoxy and lower alkanoyl groups contain 1 to3 carbon atoms. 5-amino or substituted amino 1,2, 3-triazole compounds are reacted with orotic acid to form orotate compounds of the 5-amino or substituted amino 1,2, 3-triazole compounds in a ratio ranging from 1:1 to 1:4 (base structure: acid), using the improved and safer process of the invention to form CTOs using the process according to the invention.

The novel polymorphic form of CAI is further reacted with orotic acid to form an orotate compound of the class of compounds of formula II:

formula II

Wherein the orotic acid is ionically bound to R2,

R₁is that

Wherein p is 0 to 2; m is 0 to 4; and n is 0 to 5; x is O, S, SO₂、CO、CHCN、CH₂Or C = NR₆Wherein R is₆Is hydrogen, lower alkyl, hydroxy, lower alkoxy, amino, lower alkylamino, di-lower alkylamino or cyano; and R₄And R₅Independently a halogen atom (F, Br, Cl), cyano, trifluoromethyl, lower alkanoyl, nitro, lower alkyl, lower alkoxy, carboxy, lower alkoxycarbonyl, trifluoromethoxy, acetylamino, lower alkylthio, lower alkylsulfinyl, lower alkylsulfonyl, trichloroethyl, trifluoromethylthio, trifluoromethylsulfinyl, or trifluoromethylsulfonyl; r₂Is amino, mono-or di-lower alkylamino, acetamido, acetylimino, ureido, carboxamido, iminomethyl or guanidino; and R₃Is carbamoyl, cyano, carbazolyl, amidinoOr N-hydroxycarbamoyl; wherein the lower alkyl, lower alkoxy and lower alkanoyl groups contain 1-3 carbon atoms.

Preferred embodiments of "CTO" as defined herein have empirical formula C₂₂H₁₆Cl₃N₇O₆Molecular weight 580.76, and two transition melting points (transitionMeltintPoint) at 201 ℃ and 236 ℃. CTO includes novel polymorphic forms of CAI ionically bound to orotate. CAI has multiple polymorphs, including but not limited to form1 (type 1) or form2 (type 2). Two embodiments of CTO have different transition melting points, e.g., CTO (type 1) has melting points at about 136 ℃, 194 ℃ and 235 ℃; and CTO (type 2) has melting points at about 137 ℃ and 234 ℃. Two embodiments of the CTO have an orotic acid structure consistent with CAI¹HNMR spectra (fig. 1 and 2, respectively), and FT-IR spectra consistent with form1 and form2 (fig. 3 and 4, respectively). CTO is crystalline as shown by x-ray powder diffraction patterns for form1 and form2 (fig. 5 and 6, respectively).

Chemical names for preferred embodiments of CTO include:

5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide compound with orotic acid; 5-amino-1- (3, 5-dichloro-4- (4-chlorobenzoyl) benzyl) -1H-1,2, 3-triazole-4-carboxamide compounds with orotic acid; and 5-amino-1- { [3, 5-dichloro-4- (4-chlorobenzoyl) phenyl ] methyl } -1H,1,2, 3-triazole-4-carboxamide compounds with orotic acid.

More specifically, the chemical structure of a CTO polymorph is:

CAI: orotic acid

Alternative embodiments include formulations of different polymorphs of CAI and orotic acid (CAO). Using the method according to the invention, 5-amino or substituted amino 1,2, 3-triazole (CAI) or a novel polymorph of 5-amino or substituted amino 1,2, 3-triazole is mixed with orotic acid in a ratio of 1:1 to 1:4 (basic structure: acid) to prepare a CAO formulation.

The new method comprises the following steps:

in the novel process of the present invention, the compounds of the present invention can be prepared by reaction scheme II shown below in five (5) steps. More specifically, in step 3, the novel process uses diphenyl phosphorazidate to react with intermediate 858.B instead of sodium azide. This process omits step 3 of the prior art, which forms intermediate 858. C. Refer to scheme I above (six steps). The method will be described in detail in examples. 858. A-858. F represent intermediate products and CTO as summarized below:

858.A represents tert-butyldimethylsilyl-3, 5-dichlorobenzyl ether.

858.B represents 3, 5-dichloro-4- (4-chlorobenzoyl) benzyl alcohol.

858.C represents 3, 5-dichloro-4- (4' -chlorobenzoyl) benzyl chloride.

858.D represents 3, 5-dichloro-4- (4' -chlorobenzoyl) benzyl azide.

858.E represents 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide.

858.F shows 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide compound with orotic acid, (CAI: orotic acid) (CAI: orotate) (CTO).

Importantly, it has been observed that the different CAI polymorphs, CTO and CAO prepared by the above methods show less gastric damage and toxicity in rodents when compared to CAI synthesized by the methods described in the prior art. This may be associated with the lack of use of toxic components such as sodium or potassium azide.

The novel process also enables the production of novel CAI polymorphs, CTO and CAO. Thus, the compounds of the present invention include molecules that crystallize in more than one different crystal structure, and the compounds exhibit different chemical properties of different polymorphs of CAI as determined by techniques such as NMR, DSC, FT-IR and XRDP (fig. 1-6).

Dosage and formulation

5-amino or substituted amino 1,2, 3-triazoles and substituted derivatives thereof, prepared as different polymorphs of chemical and biological nature, overcome the disadvantages of CAI produced by the processes described in the prior art.

In addition, 5-amino or substituted amino 1,2, 3-triazoles and substituted derivatives thereof have been chemically reacted with orotic acid to form orotate (CTO) salts in a ratio of 1:1 to 1:4 (basic structure: acid) with unique bioavailability, pharmacokinetic properties, safety and efficacy.

An alternative embodiment comprises combining polymorphs of 5-amino or substituted amino 1,2, 3-triazole and substituted derivatives thereof with orotic acid in a ratio of 1:1 to 1:4 (base: acid) to form a formulation of CAI and orotic acid (CAO).

The pharmaceutical compositions and formulations described above may be formulated as pharmaceutical preparations for administration to a mammal for the prevention and treatment of primary and metastatic neoplasms, chronic myeloid leukemia, macular degeneration, retinopathy and other cell proliferative disorders. The various triazole orotate compounds may be provided directly as organic acid salts or with pharmaceutically compatible counter ions, as long as the form is water soluble. Salts are more readily soluble in aqueous or other protic solvents than the corresponding free base structure form. The therapeutic compound or pharmaceutical composition may be administered intravenously, intraperitoneally, subcutaneously, intramuscularly, intrathecally, orally, rectally, topically, or by aerosol administration.

Formulations suitable for oral administration include solid powder formulations, liquid solutions of the active compound dissolved in a diluent such as saline, water or PEG 400; capsules or tablets, each containing a predetermined amount of the active agent in solid, powder, granule or gelatin form; suspensions in similar media; and an emulsion.

Formulations suitable for parenteral administration include aqueous or non-aqueous isotonic sterile solutions containing buffers, antioxidants and preservatives. The formulation may be sealed within the container in a single dose or in multiple doses.

The patient dose for oral administration of CTO ranges from 0.25 to 500 mg/day, usually 25 to 100 mg/day, and typically 50 to 400 mg/day. A typical dosage range is 0.005-10 mg/kg/day, usually 0.5-2.0 mg/kg/day, typically 1.0-8.0 mg/kg/day, expressed in terms of patient weight. A typical dosage range is from 0.1 to 300mg/m, described in terms of the patient's body surface area²A daily dose of 20 to 250mg/m²A day, typically 25-50 mg/m²The day is. The amount and spacing of the doses can be adjusted individually to provide plasma levels of the active moiety that are sufficient to maintain an anti-proliferative, anti-metastatic, anti-angiogenic effect or other therapeutic effect in diseases that depend on aberrant signal transduction and proliferation.

The dosage may be adjusted depending on the route of administration, e.g. intravenous, inhalation/aerosol, direct intraperitoneal or subcutaneous, topical or intrathecal administration.

A wide variety of delivery systems for pharmacological compounds may be utilized, including but not limited to: liposomes, nanoparticles, suspensions, and emulsions. The pharmaceutical compositions may also include suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include, but are not limited to: calcium carbonate, calcium phosphate, various sugars, starch, cellulose derivatives, gelatin, and polymers such as polyethylene glycol.

Furthermore, the drug may be administered in a targeted drug delivery system, e.g., in a liposome coated with a tumor-specific antibody, in a nanoparticle or other form. The liposomes or nanoparticles can target and be selectively taken up by a tumor or other disease target.

One of the most difficult properties to build into the newly discovered representative molecules (leadmolecules) is the required pharmacokinetic profile, especially in the case of orally administered compounds. The "most experienced pharmacist would prefer to start with a series of structures that have inherently good pharmacokinetic properties but have poor potency for the target receptor and then start to increase potency against the target rather than working in the other direction", "organic chemistry drug discovery, drug discovery", Science303: 1810-.

Enhancing bioavailability of orally administered CTO

The present invention relates generally to methods of improving oral bioavailability, delivery, and CTO (unique orotate salt of L651582 in a ratio of 1:1 to 1:4 (base structure: acid)) clearance. The present invention provides a method for preparing orotate salts of water-insoluble drugs having ionizable centers to improve the drugs' oral bioavailability, toxicological profile and efficacy. Preferably, the ratio of CTO is 1:1, and more preferably the ratio thereof is 1:2, and most preferably the ratio thereof is 0.7: 1.3.

The absorption of drugs by the oral route is the subject of intense research in the pharmaceutical industry, since good bioavailability means that the drug can reach the systemic circulation through the mouth. Oral absorption is influenced by drug properties and the physiology of the gastrointestinal tract, including dissolution of the drug from the dosage form, the mode of action of the drug with the aqueous environment and membrane, permeation through the membrane, and irreversible removal by first-pass organs such as the intestine, liver and lungs. Some drugs that exhibit low solubility exhibit poor bioavailability or irregular absorption, the degree of irregularity being affected by various factors, such as dose level, dietary status of the patient, and physicochemical properties of the drug.

Due to the large surface area, drug absorption mostly occurs in the small intestine because of the increased absorption area caused by the villi and microvilli. The intestinal circulation is unique in that the intestine is the anterior or entry tissue that regulates the flow of material to the liver. The intestinal venous blood constitutes about 75% of the blood supplied to the liver. Thus, for drugs that are cleared by the gut, the effect of the liver, kidney or lung on drug metabolism will be reduced. In contrast, for poorly extracted drugs from the intestine, the material is able to reach the next organ, the liver and lungs responsible for removal. Thus, the concentration of drug entering the intestine and intestinal flux alter the rate of drug delivery and affect the proportion in the intestine, and clearance via hepatic first-pass metabolism (hepacicfirst-passmetollism).

"drug bioavailability" is defined herein as the amount of drug available systemically over time. The present invention increases the bioavailability of a drug by converting the drug into an orotate salt. This can be achieved by modifying the hydrophilic and lipophilic properties of the drug so that the drug permeates well through the membrane and the blood flow rate becomes the overall rate-limiting step for absorption, or by inhibiting the biotransformation of the drug in the gut and/or by inhibiting the active reverse transport system (activbacktranspartstein) in the gut which reduces the net transport of the drug through the gut membrane into the blood stream. In each case, the composition that results in improved bioavailability of the drug is the orotate salt of the pharmaceutical agent. For several reasons that are not clear, it was found that the conversion of water-insoluble L651582 to CTO (basic structure: acid, 0.5: 1-1: 2) provides a means to increase the bioavailability of pharmaceutical agents orally administered to a mammal in need of treatment.

Change in integrated systemic concentration over time by area under the curve (AUC) or C_maxBoth parameters are shown as known in the art.

The present invention provides methods wherein the composition provides an increased bioavailability of the orotate salt of the pharmaceutical agent as measured by AUC of at least 25% to 100% relative to dosing of the pharmaceutical agent.

The present invention provides compositions that increase the bioavailability of orotate salts of pharmaceutical agents by at least 50% -100% as measured by Cmax.

"side effects" or "toxicity" or "adverse drug reactions" of chemotherapeutic agents are observed in the acute phase of chemotherapy administration and in patients cured of cancer with subclinical tissue damage. Potentially quite serious, disabling and irreversible drug-related tissue side effects are more highly recognized. The clinician must be aware of the potential tissue/organ complications of the chemotherapeutic agent and know to conduct an appropriate baseline tissue examination before starting treatment.

"purging" of the drug is performed by perfusing the blood to the extraction organ. "extraction" refers to the irreversible removal (excretion) or change in the ratio of the drug supplied to the organ to a different chemical form (metabolism).

The present invention provides methods for increasing the clearance of CTO orotate derivatives from non-cancerous or normal tissues by at least 25% to 100% relative to dosing with a pharmaceutical agent as measured by pharmacological studies.

By "bioavailability" of an orally administered drug is meant the degree or proportion of the active portion of the drug or metabolite that enters the systemic circulation and thereby reaches the site of action. The physicochemical properties of the drug determine its absorption capacity, as well as binding to serum proteins. The efficacy of a drug depends on its interaction with a molecular target. Thus, the nature of its dosage form depends in part on its chemical nature and the method of bulk preparation of the drug. Differences in bioavailability, potency, transport and clearance of chemical formulations of a given drug can be clinically significant.

The rate of "absorption" is important because even when the drug is completely absorbed, it may be absorbed too slowly to produce therapeutic blood levels quickly enough, or too quickly to be toxic to achieve high drug concentrations at therapeutic levels after each administration. Absorption is carried out by one of three methods, passive diffusion, active transport or assisted active transport. Passive diffusion is the simple passage of molecules across a mucosal barrier until the concentration of molecules on both sides of the membrane reaches osmotic equilibrium. In active transport, molecules are actively pumped through the mucosal layer. In assisted transport, a carrier (usually a protein) is required to transport molecules across the membrane for uptake. The present invention provides CTO compounds in a chemical configuration that allows successful drug delivery to different tissues and organs, even across the blood brain barrier to the brain.

Orotic acid, a free pyrimidine, is important in the synthesis of uridylic acid (UPP), the major pyrimidine nucleic acid. Pyrimidines play an important role in cellular regulation and metabolism. They are substrates for DNA/RNA biosynthesis, regulators of the biosynthesis of some amino acids, and cofactors in the biosynthesis of phospholipids, glycolipids, sugars, and polysaccharides. The classical pathway of nascent pyrimidine biosynthesis ends with the synthesis of UMP. Biochemistry, ed.by Lubert Stryer, W.H.Freeman & CoNY, 4 th edition, 739-762 (1995). The present invention provides a class of CTOs that undergo dissociation (dissolution) to release the drug as a charged molecule and free orotate, which can prevent binding of the drug and protein and facilitate delivery of the drug to the target, as well as rapid clearance.

Embodiments provided herein show an increase in CTO potency as measured by the following improvements: 1) the efficacy of CTO compared to an equivalent dose formulation of CAI + orotic acid, 2) the bioavailability and clearance of CTO when dosed as a solid encapsulated CTO compared to CTO in PEG-400, 3) delivery of orally administered CTO across the blood brain barrier to the brain, 4) delivery of orally administered CTO to different ocular tissues including choroid-retina complex and vitreous humor in dogs.

Importantly, pre-clinical toxicity of CTO was determined in dogs at doses of 175, 350, 1025 mg/kg/day via the PO route, and no mortality occurred after 28 days.

6. Examples of the embodiments

Example 1.

4-chlorobenzoyl chloride

3, 5-dichlorobenzyl alcohol (1 mole) was treated with tert-butyldimethylsilyl chloride (1.05 mole), imidazole, 99% (2.44 mole), 4-dimethylaminopyridine in N, N-dimethylformamide at cold temperature to produce tert-butyldimethylsilyl-3, 5-dichlorobenzyl ether (858.A1) using extraction.

Example 2

3, 5-dichloro-4- (4-chlorobenzoyl) benzyl alcohol

Tert-butyldimethylsilyl-3, 5-dichlorobenzyl ether (858.a1) (1 mole) was reacted with a 1.6M solution of n-butyllithium in hexane, then with 4-chlorobenzoyl chloride, (1.01 mole) in tetrahydrofuran, kept cold, and the intermediate was treated with aqueous hydrochloric acid to give 3, 5-dichloro-4- (4-chlorobenzoyl) benzyl alcohol (858. B).

Example 3

3, 5-dichloro-4- (4-chlorobenzoyl) benzyl azide

3, 5-dichloro-4- (4 '-chlorobenzoyl) benzyl alcohol (858.B) (1 mole) was reacted with diphenylphosphoryl azide (DPPA) (1.2 moles, and 1, 8-diazabicyclo [5.4.0] undec-7-ene, (alias: DBU) (1.2 moles)) in toluene at cold temperature followed by aqueous workup and alcohol titration to give 3, 5-dichloro-4- (4' -chlorobenzoyl) benzyl azide (858. D). DPPA is an organic compound that is used in the synthesis of other organic compounds. Aust.J.Chem26:1591-1593 (1973). DPPA was distilled at 157 ℃, and no significant nitrogen production was observed up to 175 ℃ indicating its stability to heat.

Example 4

5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-Carboxamide (CAI)

Reaction of 3, 5-dichloro-4- (4' -chlorobenzoyl) benzyl azide (858.D) (1 mole) with cyanoacetamide in hot acetonitrile (1.69 moles), and potassium carbonate, (6.2 moles) afforded 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide (858. E).

Example 5

5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxylic acid with orotic acid Amine compound, (CAI: orotic acid)

5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide (858.E) (1 mole) was reacted with orotic acid (1.03 mole) and methanol/water mixture to give a solid compound of 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide with orotic acid, (CAI: orotic acid; 1:1) (CTO) (858.F), MW580.76g, having a transition melting point at about 151 ℃, 238 ℃ and 332 ℃ as determined by differential scanning calorimetry. XRPD spectra show CTO including crystalline and amorphous (polymorphic) materials.

Example 6

Comparison of anticancer Activity of CTO (858.F) and CAI + Orotic acid (1:1)

The efficacy of CTO of molecular weight 580.8 and CAI + orotic acid of molecular weight 156.1 of molecular weight 424.6 was studied using s.c. -transplanted HT29 human colon tumor xenografts into which HT29 fragments were transplanted in male, athymic NCr-nu/nu6 week old mice and which were divided into 3 groups, ten groups, after 13 days. For the next 14 days (13-26 days), group 1 control (C) received vehicle; group 2=343 mg/kg/dose;group 3=240 mg/kg/day CAI +103 mg/kg/day orotic acid. Mean tumor volume (mm) measured on day 41³) As follows:

group 1 (control) =1436mm³

Group 2(CTO343 mg/kg/day) =864mm³(p =0.0050, Gr2 vs. Gr1)

Group 3(CAI250 mg/kg/day + orotic acid 103 mg/kg/day) =1268mm³(p =0.2706, Gr3 vs Gr 1). These results indicate that CTO is more effective in inhibiting tumor growth than comparable amounts of CAI and orotic acid, which do not chemically react between them. The CAI + orotic acid formulation, however, showed some tumor suppression.

Example 7

Comparison of orally administered CTO as a solid in a capsule or as a liquid in PEG-400

The bioavailability of CTO (basic structure: acid, 0.7:1.3) was determined by administration of a single dose of 685mg/kg with capsules (group 1) or oral gavage (group 2) in PEG 400. Two dogs (1 female/1 male) were used in each group. Blood samples were taken at 0, 1,2, 4, 8, 12, 24, 48, 72 and 92 hours. CAI was determined by HPLC/MS.

Group 1 receiving capsules: plasma concentrations after 1 hour were 155 and 174ng/ml for male and female dogs. At 12 hours, the Cmax for males was 5800ng/ml, and at 24 hours, the Cmax for females was 7950 ng/ml. Half-lives were 18 hours and 22.7 hours for males and females, respectively, and AUC values were 326 and 277 μ g/mL, respectively.

Group 2 received a gavage in PEG400: for male and female dogs, plasma concentrations after 1 hour were 511 and 570 ng/ml. At 24 hours, the Cmax for males was 6634ng/ml, and at 24 hours, the Cmax for females was 5350 ng/ml. The bioavailability was 81.8% of group 1 (100%).

These results show that administration of CTO in solid form in capsules has a better absorption pattern (absorptionpotern) and bioavailability than CTO in PEG 400. Based on these and additional results, CTO will be administered to the patient in solid form in capsules.

Example 8

CTO crossing the blood brain barrier in orally administered mice

CTO (in PEG400) was administered orally to 6 week old mice divided into two groups. Two doses were administered-group 1=513 mg/kg; group 2=342 mg/kg. After 8 hours of treatment with CTO, mice were euthanized to measure CTO concentration in brain tissue (as CAI).

The results obtained were as follows: group 1-CAI levels were 15167 ± 2372ng/g tissue; group 2 had CAI levels of 10950. + -.1704 ng/g tissue, both within the therapeutic range (6000 ng/mL). Because CTO is administered orally, these results show that CTO crosses the blood brain barrier and reaches the target organ, i.e. the brain.

The present invention is not to be limited in scope by the embodiments disclosed in the examples which are intended as illustrations of one aspect of the invention and any method which is functionally equivalent is within the scope of the invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to be included within the scope of the appended claims.

Those skilled in the art will know, or be able to ascertain using no more than routine experimentation, equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims (6)

1. A form1 polymorph of 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide bound to orotic acid having an X-ray powder diffraction pattern substantially as shown in figure 3 wherein the ratio of base structure to acid is from 1:1 to 1: 4.

2. A form2 polymorph of 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide bound to orotic acid having an X-ray powder diffraction pattern substantially as shown in figure 4, wherein the ratio of base structure to acid is from 1:1 to 1: 4.

3. A process for the manufacture of form1 polymorph of orotic acid bonded 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide as claimed in claim 1 comprising:

4. a process for the manufacture of form2 polymorph of orotate-bound 5-amino-1- (4- (4-chlorobenzoyl) -3, 5-dichlorobenzyl) -1,2, 3-triazole-4-carboxamide of claim 2, comprising:

5. use of the polymorph of claim 1 or 2 or manufactured by the process of claim 3 or 4 for the preparation of a medicament.

6. Use of the polymorph of claim 1 or 2 or manufactured by the process of claim 3 or 4 for the preparation of a medicament for the treatment of solid cancer, macular degeneration, retinopathy, chronic myeloid leukemia or AIDS.