TWI686197B - Solid pharmaceutical compositions comprising biopterin derivatives and uses of such compositions - Google Patents

Abstract

The present invention relates to solid pharmaceutical compositions comprising biopterin derivatives as well as methods for obtaining such solid pharmaceutical compositions. The invention also relates to the solid pharmaceutical compositions of the invention for treating diseases.

Description

Solid pharmaceutical compositions containing biological pterin derivatives and these combinations Use of things

The present invention relates to solid pharmaceutical compositions containing biopterin derivatives and methods for obtaining such solid pharmaceutical compositions. The invention also relates to the treatment of diseases with the solid pharmaceutical composition of the invention.

Biopterines and their derivatives are molecules of medical interest. For example, tetrahydrobiopterin (BH4, sapropterin) has recently been approved for the treatment of hyperphenylalanineemia (HPA) in adults and four-year-old children, as well as phenketonuria ( phenylketonuria; PKU). For this purpose, tetrahydrobiopterin is formulated as a soluble tablet commercial drug (sold under the name Kuvan), which contains mannitol (E421), anhydrous calcium hydrogen phosphate, and crospovidone type A (Crospovidone type A ), ascorbic acid (E300), sodium stearyl fumarate, and riboflavin (E101). See also the full text of WO 2006/055511.

Other therapeutically promising biological pterin compounds are 4-amino-5,6,7,8-tetrahydro-L-biopterin and 4-amino-7,8-dihydro-L-biopterin . Both compounds show properties that are different from other nitric oxide inhibitors, making these compounds more suitable than "typical" arginine analogs (Werner et al al., (1996). Biochemical Journal 320,93-6 or US Patent No. 5,922,713). Two compounds have shown utility in experimental TBI (see for example, WO 2004/084906, US Patent No. 8,222,828, European Patent No. 0 906 913, or Terpolilli et al., J Neurotrauma. 2009; 26(11): 1963-75 ). However, their biological pterin derivatives have not been approved for medical treatment. Therefore, it is necessary to provide pharmaceutical compositions suitable for human therapeutic applications. Ideally, their pharmaceutical compositions should be easy to prepare, easy to use, and stable. It should be noted that when stored for a long time or supplied as a solution, the hydrogenated biopterin derivatives are sensitive to oxidation.

The object of the present invention is therefore to provide a pharmaceutical composition comprising a bioptopterin derivative which meets these needs.

This problem is solved by specific examples of the present invention, such as the definition of the scope of patent application, the description of the implementation, and the description of the examples and drawings.

及/或式(II)化合物：

以及 b)至少一磷酸鹽。 A specific embodiment of the present invention relates to a solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate.

The solid composition of the present invention may be "suitable for intravenous administration". This refers to the combination of a solid composition and a pharmaceutically acceptable carrier, preferably a pharmaceutically acceptable fluid, such as water or buffer or any reconstitution fluid, to obtain the composition, which is then used directly Intravenous application. Therefore, after mixing the solid composition with a pharmaceutically acceptable carrier, a ready-made composition is obtained, which is suitable for intravenous application.

In a specific embodiment of the pharmaceutical composition of the present invention, the at least one phosphate is sodium phosphate, potassium phosphate, or ammonium phosphate. Phosphate can be selected from Na ₂ HPO ₄ (anhydrous), Na ₂ HPO ₄ ˙2 H ₂ O, Na ₂ HPO ₄ ˙7 H ₂ O, Na ₂ HPO ₄ ˙12 H ₂ O, NaH ₂ PO ₄ ( Anhydrous), NaH ₂ PO ₄ ˙H ₂ O, NaH ₂ PO ₄ ˙2 H ₂ O, K ₂ HPO ₄ (anhydrous), K ₂ HPO ₄ ˙3 H ₂ O, KH ₂ PO ₄ (anhydrous), and Group of mixtures.

In some embodiments of the pharmaceutical composition of the present invention, the phosphate is Na ₂ HPO ₄ ˙2 H ₂ O, and the amount of Na ₂ HPO ₄ ˙2 H ₂ O present in the composition is selected so that Na ₂ The molar ratio of HPO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.04 to 0.4.

In a further embodiment of the pharmaceutical composition of the present invention, sodium phosphate is NaH ₂ PO ₄ ˙ ₂ H ₂ O, and the amount of NaH ₂ PO ₄ ˙ ₂ H ₂ O present in the composition is selected so that NaH ₂ The range of the molar ratio of PO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.01 to 0.09.

In other specific embodiments, the pharmaceutical composition of the present invention includes two different sodium phosphate salts. Arbitrarily, the two different sodium phosphate salts are NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O.

In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O present in the composition is selected so that NaH ₂ PO ₄ 2 The molar ratio of H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O to compound (I) or compound (II) is in the range of 0.02 to 0.5.

In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O present in the composition is selected so that NaH ₂ PO ₄ 2 The range of the molar ratio of each of H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.02 to 0.5.

In a further specific embodiment of the pharmaceutical composition of the present invention, compound (I) and/or compound (II) are present as free bases.

In some embodiments of the pharmaceutical composition of the present invention, the pharmaceutical composition is a freeze-dried pharmaceutical composition.

In another specific embodiment of the pharmaceutical composition of the present invention, compound (I) is a compound of formula (Ia):

In another specific embodiment of the pharmaceutical composition of the present invention, compound (II) is a compound of formula (IIa):

In some embodiments of the pharmaceutical composition of the present invention, the pharmaceutical composition includes additional pharmaceutical excipients.

In another specific embodiment of the pharmaceutical composition of the present invention, the additional pharmaceutical excipient is an inorganic salt. The inorganic salt may be selected from MgCl ₂ , CaCl ₂ , NH ₄ Cl, KCl, or NaCl, preferably the inorganic salt is NaCl.

In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaCl present in the composition of the present invention is selected so that the molar ratio of NaCl to compound (I) or compound (II) ranges from 1.5 to 4, Preferably 1.8 to 3.7.

In some embodiments of the pharmaceutical composition of the present invention, it further comprises crystal water.

In other specific embodiments, the pharmaceutical composition of the present invention is suitable for reconstitution in water. "Suitable for reconstitution in water" means that the composition (after dehydration or concentration) can be restored to a liquid state by adding water.

In a further specific embodiment, the pharmaceutical composition of the present invention is suitable for administration by infusion or injection.

In some specific embodiments of the pharmaceutical composition of the present invention, compound (I) is (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

In a further specific embodiment of the pharmaceutical composition of the present invention, compound (I) is (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

In other specific embodiments of the pharmaceutical composition of the present invention, compound (I) is a mixture of diastereomers, which contains (6R)-4-amino-5,6,7,8-tetrahydro-L -Biopterin is more than (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

In some specific embodiments of the pharmaceutical composition of the present invention, (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino-5, The amount of 6,7,8-tetrahydro-L-biopterin is selected so that (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S )-4-Amino-5,6,7,8-tetrahydro-L-biological butterfly, the ratio of the ratio is 0.5 to 2, preferably about 1.3.

In a further embodiment of the pharmaceutical composition of the present invention, the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biological butterfly Free base, 140±30 mg of crystal water, 70±7 mg of disodium hydrogen phosphate dihydrate (Na ₂ HPO ₄ .2 H ₂ O), 16.5±2 mg of sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ .2 H ₂ O), and 350±30mg of sodium chloride (NaCl).

In other specific embodiments of the pharmaceutical composition of the present invention, the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin the free base of 60 ± 50mg crystalline water, disodium hydrogen phosphate dihydrate of 70 ± 7mg _{(Na 2 HPO 4 .2 H 2} O), 12 ± 2.5mg of sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ .2 H ₂ O), and 350 ± 30mg of sodium chloride (NaCl).

In some embodiments of the pharmaceutical composition of the present invention, the composition comprises one, two, three, four, five, six, seven or more additional compounds, wherein the additional compound is selected from one of the compounds Or a group of multiples, the compound is selected from 4-amino-L-biopterin, (6R,S)-5,6,7,8-tetrahydro-L-biopterin, 1 -[(6R,S)-2,4-diamino-5,6,7,8-tetrahydropyridin-6-yl]propanol, 1-[(6R,S)-2,4-di Amino-5,6,7,8-tetrahydropyridin-6-yl]propane, (1R,2S)-1-[(6R,S)-2-(ethylaminoamino)-4-amine Group-5,6,7,8-tetrahydropterin-6-yl)-1,2-acetoxypropane, 2,4-diamino-7,8-dihydropteridine, 2,4 -A group consisting of diaminopyridines.

The invention also relates to the use of the freeze-dried pharmaceutical composition of the invention to treat diseases. In some embodiments, the disease is selected from the group consisting of traumatic brain injury, non-traumatic brain injury, preferably stroke or meningitis, elevated cerebral pressure, and secondary brain injury.

The invention also relates to freeze-dried pharmaceutical compositions for the treatment of diseases. In some embodiments, the disease is selected from the group consisting of traumatic brain injury, non-traumatic brain injury, preferably stroke or meningitis, elevated cerebral pressure, and secondary brain injury.

及/或式(II)化合物：

以及b)至少一磷酸鹽，及任意地NaCl；該方法包含：aa)以緩衝液溶解式(III)化合物及/或式(II)化合物：

其中較佳地緩衝液包含磷酸鹽；bb)凍乾aa)取得之溶液。 The present invention further relates to a method for preparing a lyophilized solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate, and optionally NaCl; the method comprises: aa) dissolving the compound of formula (III) and/or the compound of formula (II) in a buffer:

Preferably, the buffer solution contains phosphate; bb) lyophilized aa) the obtained solution.

In some embodiments, the method of the present invention further comprises the step of dissolving the lyophilizate obtained in bb) in a pharmaceutically acceptable fluid to prepare an injection solution.

及/或式(II)化合物：

以及b)至少一磷酸鹽，及任意地NaCl；該方法包含：aa)以緩衝液溶解式(III)及/或(II)化合物：

其中較佳地緩衝液包含磷酸鹽；bb)凍乾aa)取得之溶液；cc)將bb)取得之凍乾物重組於醫藥上可接受之流體，以製備注射液，其中bb)取得之凍乾物係填充至小瓶中。 Accordingly, the present invention also relates to a method for preparing an injection, which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate, and optionally NaCl; the method comprises: aa) dissolving the compound of formula (III) and/or (II) with a buffer:

Preferably the buffer contains phosphate; bb) lyophilized aa) obtained solution; cc) reconstituted bb) obtained lyophilisate into a pharmaceutically acceptable fluid to prepare an injection solution, wherein bb) obtained lyophilisate Fill into vials.

In a specific embodiment of the method of the present invention, bb) the obtained lyophilisate is filled into a 50ml vial, (i) preferably a solid formulation with an amount of about 1-1.5g, preferably 1.25g; or (ii) Preferably its amount is about 0.9-1.4g, preferably 1.15g of solid formula.

In other specific embodiments of the method of the present invention, the buffer of aa) is a sodium hydrogen phosphate buffer, which includes at least one phosphate.

In other specific embodiments of the method of the present invention, the buffer of aa) includes NaOH, sodium hydrogen phosphate buffer, and water. Arbitrarily, NaOH is a 5N NaOH solution.

In another specific embodiment of the method of the present invention, the sodium hydrogen phosphate buffer is prepared by dissolving NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O, respectively.

In a further specific embodiment of the method of the present invention, the sodium hydrogen phosphate buffer is pH 7.4, which is obtained by dissolving NaH ₂ PO ₄ ˙ ₂ H ₂ O in Na ₂ HPO ₄ ˙ ₂ H ₂ O solution.

In some embodiments of the method of the present invention, the buffer solution includes 12-16% (w/w) 5N NaOH, 8-12% (w/w) sodium hydrogen phosphate buffer, and 74-78% (w/w) Water for Injection.

In another embodiment of the method of the present invention, the solution obtained by aa) is sterile filtered, preferably using a 0.22 μm filter membrane.

In a further embodiment of the method of the present invention, the pH of the buffer is about 8, 9, 10, 11, 12, 13, or 14.

In yet another specific embodiment of the method of the present invention, the pH of the solution in step aa) is about 4, 5, 6, 7, 8, 9, 10, or 11, preferably between 6.5 and 7.6, the most Goodland 7.4.

In other specific embodiments of the method of the present invention, bb) the obtained lyophilisate is filled into a vial, preferably a solid formulation with an amount of about 1-1.5 g, preferably 1.25 g; or preferably an amount of about 0.9- 1.4g, preferably 1.15g of solid formula.

In a further specific embodiment of the method of the present invention, the buffer is prepared with degassed buffer. In some embodiments, the buffer solution is degassed with nitrogen until the oxygen content is less than 1.0 ppm.

In another specific embodiment of the method of the present invention, after the solution is prepared, the lyophilization method starts at most 2 hours later.

The invention also relates to the pharmaceutical composition obtained by the method of the invention.

The present invention further relates to the use of the lyophilized pharmaceutical composition of the present invention as a medicament for the treatment of traumatic brain injury such as intracranial injury, increased cerebral pressure and secondary brain injury, or non-traumatic Individuals with brain damage such as stroke or meningitis.

The present invention also relates to a method of treating a disease of an individual, which includes the step of administering the lyophilized pharmaceutical composition of the present invention to an individual in need. One of the methods for treating individual diseases In some specific embodiments, the maximum daily dose is 20 mg/kg body weight/day, preferably, 17.5, 15.0 or 12.5, 10, 8.5, 7.5, 5.0, or 2.5 mg/kg body weight/day.

The present invention also relates to the use of a lyophilized pharmaceutical composition to treat an individual's disease. The use includes the step of administering the lyophilized pharmaceutical composition of the present invention to an individual in need. In some embodiments, the maximum daily dose is 20 mg/kg body weight/day, preferably 17.5, 15.0 or 12.5, 10, 8.5, 7.5, 5.0, or 2.5 mg/kg body weight/day.

及/或式(II)化合物：

以及b)至少一無機鹽，較佳地NaCl。 In yet another specific embodiment, the present invention relates to a solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one inorganic salt, preferably NaCl.

In this specific embodiment, the amount of NaCl present in the composition of the present invention is selected so that the molar ratio of NaCl to compound (I) or compound (II) is 1.5 to 4, preferably 1.8 to 3.7 , 1.85 to 3.6, 1.9 to 3.4, optimally 1.9 to 2.5.

In a specific embodiment, the molar ratio of NaCl to compound (I) or compound (II) is about 2.2.

Figure 1 shows the preparation of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride dihydrate (VAS203) drug substance.

Figure 2 shows the manufacturing process of the VAS203 drug product of the present invention.

Figure 3 shows the storage stability of 1 g VAS203 vial (Lot No. 928606) at 2-8°C (Table 5).

The object of the present invention is to provide a pharmaceutical composition. In addition, these compositions should be easy to use and stable. It is found in this article that by using solid phosphates (alone or in combination with inorganic salts such as NaCl or KCl), and in combination with solid biological pterin derivatives such as compounds of formula (I) or (II), pharmaceutical compositions (formulations) are produced, which are extremely It is suitable for a certain pH value or pH range and osmotic pressure, and when dissolved, it is directly suitable for therapeutic administration. The solid pharmaceutical composition is particularly suitable and easy to use because it is suitable for intravenous administration, thereby ensuring safe use. In addition, by providing a solid composition, the problem of oxidation of biological pterin derivatives in the liquid is solved.

及/或式(II)化合物：

以及b)至少一磷酸鹽。 Therefore, the present invention relates to a solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate.

及/或式(II)化合物：

以及b)至少一無機鹽，具體地NaCl。 In addition, the present invention also relates to a solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one inorganic salt, specifically NaCl.

The words "solid" or "solid composition" as used herein refer to particles (ions, atoms or molecules, or compounds), which are closely packed together. The force between the particles is so strong that the particles cannot move freely and can only vibrate. As a result, the solid has stability, exact shape, and exact volume. A solid can only change its shape with external force, such as breaking or cutting. For crystalline solids, the particles (atoms, molecules, or ions) are arranged in a regular sequence and repeating patterns. There are various crystalline structures, and the same substance can have more than one structure (or solid phase). The term "solid" also covers amorphous or amorphous compositions/substances/solids. Usually, the agglutination state is measured at room temperature and ambient pressure. Room temperature often represents a small temperature range, where the air feels neither cold nor hot, often indicating a temperature range between 20 and 23.5°C, with an average of 21°C (70°F). Ambient pressure means that the pressure is between 900 and 1200hPa (HPa), preferably about 1000hPa. The solid can be transformed into a liquid by melting, and the liquid can be transformed into a solid by freezing. Solids can also be directly converted into gases through the sublimation process.

The solid composition of the present invention can be administered (usually dissolved in a liquid such as water) to an individual ("administered"). This is to provide an individual with a therapeutically effective dose of the solid composition of the present invention.

"Therapeutically effective amount" is the dose of the compound of formula (I) and/or formula (II), which produces an effect when administered. The precise dosage depends on the purpose of the treatment and can be ascertained by those skilled in the art using conventional techniques. As is known in the art and described above, it may be necessary to adjust systemic and local delivery, age, weight, overall health, gender, diet, time of administration, drug interaction, and severity of the condition, and those skilled in the art can routinely The experiment confirmed. In this article, it should be noted again that the two compounds, namely 4-amino-5,6,7,8-tetrahydro-L-biopterin and 4-amino-7,8-dihydro-L-biological Pterin is active in medicine. It should also be noted that 4-amino-7,8-dihydro-L-biopterin can be oxidized by 4-amino-5,6,7,8-tetrahydro-L-biopterin (and spontaneously Sexual oxidation). Therefore, the composition of the present invention may contain only 4-amino-5,6,7,8-tetrahydro-L-biopterin or 4-amino-7,8-dihydro-L-biopterin, Or a mixture of the two compounds in any ratio.

The solid composition of the present invention can be applied to human treatment and veterinary applications. The compounds described herein having the desired therapeutic activity can be administered to the patient/individual in a pharmaceutically acceptable carrier, as described herein. Depending on the method of introduction, the compound can be formulated in a variety of ways, as described below. The solid composition of the present invention can be administered alone or in combination with other treatments.

The solid composition of the present invention can be further "suitable for intravenous administration". This means that the solid composition is preferably mixed with a pharmaceutically acceptable carrier, preferably a pharmaceutically acceptable fluid, such as water or buffer, or any of the aforementioned reconstituted fluids, to obtain the composition after mixing. It is therefore directly used for intravenous injection applications. Therefore, after mixing the solid composition with a pharmaceutically acceptable carrier, a ready-made composition is obtained, which is suitable for intravenous application. Intravenous fluid The substance is directly infused or injected into a vein, often using a syringe and hollow needle, which penetrates the skin to a sufficient depth to administer the material into the individual.

Therefore, in a further specific embodiment, the pharmaceutical composition of the present invention is suitable for administration by infusion or injection. "Infusion" refers to continuous administration for a specific period of time. For example, this administration can take between 10 minutes and 4 days. Therefore, it lasts at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 , 24, 32, 40, 48, 56, 68, 72, 86, or 96 hours. "Injection" refers to a short infusion method of putting fluid into an individual. This administration usually takes less than 10 minutes. However, the injection can be repeated multiple times a day. For example, the injection can be performed 1, 2, 3, 4, 5, 6, 7, 8, or 9 times a day. In addition, injections can be administered for 1, 2, 3, or 4 days. However, if necessary, administration via injection or infusion may take longer. Obviously, the exact time depends on many factors.

Typically, the composition for intravenous administration is a solid pharmaceutical composition of the present invention mixed with a pharmaceutically acceptable carrier, for example, a sterile isotonic aqueous buffer is mixed to form a pharmaceutical solution. If necessary, this composition/solution may also include a solubilizer and a local anesthetic such as lignocaine to relieve pain at the injection site. In general, the ingredients are supplied individually or mixed together in unit dosage form, for example, to dry lyophilized powder or anhydrous concentrate, which is placed in airtight containers such as vials, ampoules, or sachets, Also indicate the amount of compound (I) and/or compound (II). When the composition/solution is to be administered by infusion, it can be prepared with an infusion bottle containing sterile pharmaceutical grade water or saline. When the composition is to be administered by injection or infusion, an ampoule of sterile water for injection or saline solution can be provided so that the ingredients can be mixed before administration.

The term "pharmaceutically acceptable" means approved by a regulatory agency or other generally recognized pharmacopoeia for use in animals, and more specifically in humans. Generally speaking, "medically acceptable" also means "physiologically acceptable", meaning that the fluid/carrier system conforms or is characterized to function normally in vivo, It does not cause toxicity or any other adverse effects. This fluid/carrier normally has about the same pH and/or osmotic pressure as a fluid such as animal blood.

"Pharmaceutically acceptable fluid" can be one of the following non-limiting lists of non-aqueous or aqueous solvents. Non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous solvents include water, alcoholic/aqueous solutions, emulsions, or suspensions, including saline and buffered media, sodium ion solutions, Ringer's dextrose, glucose and sodium ion solutions, lactated Ringer's solution, or not Fixed oils. Intravenous injection vehicles include fluid and nutritional supplements, electrolyte supplements (such as Ringer's glucose-based fluids), and the like. Preferred aqueous solvents are pharmaceutically acceptable flow systems, such as sterile water for injection.

In some embodiments, the solid pharmaceutical composition of the present invention is reconstituted before administration. In other specific embodiments, the pharmaceutical composition of the present invention is suitable for reconstitution in water. "Recombination in water" refers to returning the composition (dehydrated or concentrated) to a liquid state by adding the aforementioned pharmaceutically acceptable fluid. Preferably, the solid composition of the present invention is reconstituted in water.

The present invention may include different solid compositions. For example, the solid composition may include compound (I) and/or compound (II) in addition to phosphate. Usable "phosphate" refers to any conventional phosphate. Phosphate refers to many different combinations of chemical phosphates with salts and minerals. In a specific embodiment of the pharmaceutical composition of the present invention, the at least one phosphate is sodium phosphate, potassium phosphate, or ammonium phosphate. Phosphate can be selected from Na ₂ HPO ₄ (anhydrous), Na ₂ HPO ₄ ˙2 H ₂ O, Na ₂ HPO ₄ ˙7H ₂ O, Na ₂ HPO ₄ ˙12 H ₂ O, NaH ₂ PO ₄ (anhydrous ), NaH ₂ PO ₄ ˙H ₂ O, NaH ₂ PO ₄ ˙2 H ₂ O, K ₂ HPO ₄ (anhydrous), K ₂ HPO ₄ ˙3 H ₂ O, KH ₂ PO ₄ (anhydrous), and mixtures thereof Formed into groups.

In some embodiments of the pharmaceutical composition of the present invention, the phosphate is Na ₂ HPO ₄ ˙2 H ₂ O, and the amount of Na ₂ HPO ₄ ˙2 H ₂ O present in the composition is selected so that Na ₂ The range of the molar ratio of HPO ₄ 2H ₂ O to compound (I) or compound (II) is 0.04 to 0.4, preferably 0.05 to 0.35, 0.075 to 0.25, 0.09 to 0.2. In a specific embodiment, the molar ratio of Na ₂ HPO ₄ 2H ₂ O to compound (I) or compound (II) is about 0.144.

In a further embodiment of the pharmaceutical composition of the present invention, sodium phosphate is NaH ₂ PO ₄ ˙ ₂ H ₂ O, and the amount of NaH ₂ PO ₄ ˙ ₂ H ₂ O present in the composition is selected so that NaH ₂ The range of the molar ratio of PO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.01 to 0.09, preferably 0.015 to 0.07, 0.02 to 0.05, 0.025 to 0.04, 0.035 to 0.05. In a specific embodiment, the molar ratio of NaH ₂ PO ₄ ˙2 H ₂ O to compound (I) or compound (II) is about 0.038.

In other specific embodiments, the pharmaceutical composition of the present invention includes two different sodium phosphate salts. Arbitrarily, two different sodium phosphate salts are NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O.

In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O present in the composition is selected so that NaH ₂ PO ₄ 2 The molar ratio of H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.02 to 0.5, preferably 0.03 to 0.45, 0.04 to 0.3, 0.05 to 0.25 , 0.06 to 0.2, 0.07 to 0.15. In a specific embodiment, the molar ratio of both NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O to compound (I) or compound (II) is about 0.18.

In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaH ₂ PO _{4 2} H ₂ O and Na ₂ HPO _{4 2} H ₂ O present in the composition is selected so that NaH ₂ PO ₄ 2 The range of the molar ratio of each of H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.02 to 0.5, preferably 0.025 to 0.4, 0.025 to 0.3, 0.025 To 0.2, 0.03 to 0.1. In some embodiments, the amount of NaH ₂ PO ₄ ˙ ₂ H ₂ O present in the composition is selected so that the molar ratio of NaH ₂ PO ₄ ˙ ₂ H ₂ O to compound (I) or compound (II) The range is 0.01 to 0.09, and the amount of Na ₂ HPO ₄ ˙2 H ₂ O present in the composition is selected so that the difference between Na ₂ HPO ₄ ˙ ₂ H ₂ O and compound (I) or compound (II) The ear ratio ranges from 0.04 to 0.4. Here, other amounts of the aforementioned phosphates Na ₂ HPO ₄ ˙2 H ₂ O and NaH ₂ PO ₄ ˙2 H ₂ O applied separately are also applicable. In some embodiments, the solid pharmaceutical composition comprises at least one, two, three, four, five, six or more phosphates. In a specific embodiment, the solid pharmaceutical composition of the present invention comprises bis (different) phosphate.

The term "about" as used herein is understood to mean that individual values or ranges (such as pH, concentration, percentage, molar concentration, time, etc.) are variable and can reach 5%, up to 10%, up to 15%, Or up to and including the specified value of 20%. For example, if the formulation contains about 5 mg/ml of compound, it is understood that the formulation may have between 4 and 6 mg/ml, preferably between 4.25 and 5.75 mg/ml, more preferably Between 4.5 and 5.5 mg/ml, and even more preferably between 4.75 and 5.25 mg/ml, with the best being 5 mg/ml. The same applies to the molar concentration. For example, a molar concentration of about 1 can be understood as a molar concentration between 0.8 and 1.2, preferably between 0.85 and 1.15, more preferably between 0.9 and 1.1, and even better Between 0.95 and 1.05, of which the best is 1.

Alternatively, the present invention relates to a solid composition comprising compound (I) and/or compound (II), and an inorganic salt other than phosphate. In these compositions, the inorganic salt may be only components other than compound (I) or (II) (in this case, no phosphate is present). Alternatively, in the composition of the present invention, the inorganic salt may exist in combination with at least one phosphate. The term "inorganic salt" as used herein refers to each applicable inorganic salt. Optionally, the inorganic salt is selected from MgCl ₂ , CaCl ₂ , NH ₄ Cl, KCl, or NaCl. In some preferred embodiments, the inorganic salt is NaCl. In some specific embodiments of the pharmaceutical composition of the present invention, the amount of NaCl present in the composition of the present invention (alone or together with at least one phosphate) is selected so that NaCl and compound (I) or compound (II) The molar ratio ranges from 1.5 to 4, preferably 1.8 to 3.7, 1.85 to 3.6, 1.9 to 3.4, and most preferably 1.9 to 2.5. In one of these specific embodiments, the molar ratio of NaCl to compound (I) or compound (II) is about 2.2.

In the composition of the present invention, compound (I) and/or compound (II) may be a mixture of diastereomers or one, two, three, four, five, six, seven, or eight stereos of compound (I) A mixture of isomers and one, two, three, four, five, or six stereoisomers of compound (II), preferably one or two stereoisomers.

The compound of formula (I) may thus comprise a mixture of diastereomers of the compound of formula (I) or a mixture of one or more stereoisomers of the compound of formula (I). Compound (I) may be (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin. In a further specific embodiment, compound (I) is (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin. In other specific embodiments, compound (I) is a mixture of diastereomers, which contains more (6R)-4-amino-5,6,7,8-tetrahydro-L-biological butterfly than ( 6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin. In some specific embodiments, (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino-5,6,7,8-tetra The amount of hydrogen-L-biopterin is selected so that (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino- The ratio of the amounts of 5,6,7,8-tetrahydro-L-biopterin is 0.5 to 2, preferably 0.5 to 1.9, 0.7 to 1.8, 0.8 to 1.7, 0.9 to 1.6, 1 to 1.5, Optimally 1.1 to 1.4. In a specific embodiment, (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino-5,6,7,8-tetra The amount of hydrogen-L-biopterin is selected so that (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino- The ratio of the amounts of 5,6,7,8-tetrahydro-L-biopterin is about 1.3. In some specific embodiments, the pharmaceutical composition of the present invention includes only compound (I).

Compound (I) may also be a compound of formula (Ia):

Similarly, the compound of formula (II) referred to herein may include a mixture of stereoisomers of the compound of formula (II) or a mixture of one or more stereoisomers of the compound of formula (II). In some specific embodiments of the pharmaceutical composition of the present invention, compound (II) is 4-amino-7,8-dihydro-L-biopterin. In another specific embodiment, compound (II) is a compound of formula (IIa):

In some specific embodiments, the pharmaceutical composition of the present invention comprises compound (II) only.

In the solid pharmaceutical composition, the compound (I) and/or the compound (II) may also exist as a free base. The "free base" referred to herein refers to the pure base form of the amine, which is relative to its salt form. In addition, the term is used to describe the deprotonated amine form of the compound. Common relative ions are inorganic acid ions, such as negatively charged chloride. For example, when HCl is added, the free base amine (NH ₂ ) is compared with the amine hydrochloride (NH ₃ ⁺ Cl ⁻ ).

As mentioned previously, the bioptopterin derivative is sensitive to liquid oxidation. Therefore, the solid pharmaceutical composition can also be supplied as a lyophilized pharmaceutical composition. The term "lyophilized" used in this article refers to freeze-drying, which is a dehydration process. Freeze-drying is by freezing the material and then reducing the surrounding pressure, so that the frozen water in the material directly sublimates from the solid phase to the gas phase. In some embodiments, the final residual water content of the lyophilized product is between 0% and 15%, preferably between 0% and 12% (w/w). The freeze-drying method is known to those skilled in the art.

As is apparent to those skilled in the art, the pharmaceutical composition may further include one, two, three, four, five, six, seven, eight, nine, ten, or more additional pharmaceutical excipients. "Pharmaceutical excipients" or additives are compounds added to compounds of formula (I) and/or (II). Such additives can serve specific functions. Its addition can increase the capacity, assist the manufacturing process, improve stability, improve drug delivery and targeting, and improve drug stability or pharmacokinetic properties. Ingredients that are used during the manufacture of pharmaceutical products but may not be present in the solid composition of the present invention are also considered as excipients (examples include water used to freeze-dry the product, and inert gas in the headspace of the container). Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, trehalose, mannitol, sorbitol, glycine, Histidine, raffinose, gelatin, malt, rice, flour, chalk, silicone, sodium stearate, glyceryl monostearate, talc, sodium ions, skimmed milk powder, glycerin, propylene, ethylenedioxide Alcohol, glucose, dextran, water, ethanol, and the like. If desired, the composition may also contain a small amount of wetting or emulsifying agents, or pH buffering agents. Such compositions can take the form of solutions, suspensions, emulsions, and the like.

For example, if the solid composition of the invention comprises a compound of formula (I) and/or formula (II) and a phosphate, a possible additional pharmaceutical excipient may be an inorganic salt. The inorganic salt may be any of the aforementioned inorganic salts. On the other hand, if the pharmaceutical composition of the present invention comprises a compound of formula (I) and/or formula (II) and an inorganic salt, the additional pharmaceutical excipient may be phosphate. The phosphate can be any of the aforementioned phosphates.

In addition to the fact that pharmaceutical excipients may be present in the solid pharmaceutical composition of the present invention, in some specific embodiments of the pharmaceutical composition of the present invention, crystal water may be included, and of course additional pharmaceutical excipients may also be present. Used herein, according to its conventional meaning in the art, "crystal water" or "hydrated water" or "crystal water" in the present invention means water that appears inside the crystal, inorganic salts (such as Na ₂ HPO ₄ or Both NaH ₂ PO ₄ ) and the biological pterin derivative of compound (I) or compound (II) may have crystal water. Although for solid phosphates such as Na ₂ HPO ₄ . 2 H ₂ O or NaH ₂ PO ₄ . In 2 H ₂ O, the crystalline water exists in a defined stoichiometric amount and the amount of crystalline water present in the solid form of compound (I) or compound (II) is variable and depends on the solid used to prepare the invention The synthesis and/or crystallization conditions of the compound of the formula. Without wishing to be bound by theory, it is believed that the crystal water present in the solid form of compound (I) or compound (II) may, for example, be bonded to the diamine group of compound (I) or (II) via hydrogen bonding, which is Free base exists. For illustrative purposes, the unit dose of the composition throughout the text contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin free base, 140±30 mg Crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 16.5±2mg NaH ₂ PO ₄ . 2 H ₂ O, and 350±30mg sodium chloride (NaCl). In this unit dose, the sodium diphosphate salt is present as a dihydrate, and the crystal water present in an amount of 140 ± 30 mg means only the water system present and the compound (I) and/or compound (II) Free base binding. Another exemplary unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin free base, 60±50 mg of crystals Water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 12±2.5mg NaH ₂ PO ₄ ． 2 H ₂ O, and 350±30mg sodium chloride (NaCl).

As mentioned, the pharmaceutical composition of the present invention can be supplied in unit doses. The "unit dose" of the solid pharmaceutical composition of the present invention means that the ingredients are mixed together in a unit dose, which typically involves active pharmaceutical components such as compound (I) and/or compound (II) and non-pharmaceutical components (excipients) Agent) such as a mixture of at least one phosphate and/or at least one inorganic salt. In addition, the unit dose may contain further excipients, such as at least one phosphate or at least one inorganic salt, and/or other non-reusable materials, which may not be considered as ingredients or packaging (such as eg capsule shells) . The term "unit dose" can also cover non-reusable packaging (especially when each drug product is packaged separately). The unit dose may also contain the reconstituted solid pharmaceutical composition of the present invention.

For an exemplary unit dose, the calculation of the molar ratio of this unit dose component is summarized below.

n Molar ratio between _NACl /n _{compound (i)} =0,005989mol/0,0027053mol~2.214

the molar ratio between the n _NACl / n _{compound (ii) = 0,005989mol / 0,0027282mol ~} 2.195

n _{Na2HPO4˙2 H2O} +n _{NaH2PO4˙2 H2O} /n mole ratio between _{compound (i)} =0.0003932mol+0.0001057mol/0,0027053mol~0.18

n _{Na2HPO4˙2 H2O} +n _{NaH2PO4˙2 H2O} /n molar ratio between _{compound (ii)} =0.0003932mol+0.0001057mol/0,0027282mol~0.18

n _{Na2HPO4˙2 H2O} /n mole ratio between _{compound (i)} =0.0003932mol/0,0027053mol~0.145

n _{Na2HPO4˙2 H2O} /n molar ratio between _{compound (ii)} =0.0003932mol/0,0027282mol~0.144

n _{NaH2PO4˙2 H2O} /n molar ratio between _{compound (i)} =0.0001057mol/0,0027053mol~0.038

n _MoH ratio of _{NaH2PO4˙2 H2O} /n _{compound (ii)} =0.0001057mol/0,0027282mol~0.038

n _{compound (i)} = m _{compound (i)} M _{compound (i)} =0,710g/240.27g mol ^-1 =0,002955mol

n _{Compound (i)} = m _{Compound (i)} / M _{Compound (i)} = 0.590g/240.27g mol ^-1 = 0.0024556mol

n _{compound (i)} = m _{compound (i)} /M _{compound (i)} = 0.650g/240.27g mol ^-1 = 0.00270553mol

n _{compound (ii)} = m _{compound (ii)} /M _{compound (ii)} = 0.710 g/238.25 g mol ^-1 = 0.0000 mol

n _{compound (ii)} = m _{compound (ii)} /M _{compound (ii)} = 0.590g/238.25g mol ^-1 = 0.00247663mol

n _{compound (ii)} = m _{compound (ii)} /M _{compound (ii)} = 0.650 g/238.25 g mol ^-1 = 0.002272282 mol

n _NACl =m _NACl /M _NACl =0,380g/58.44g mol ^-1 =0,0065023mol

n _NACl =m _NACl /M _NACl =0,320g/58.44gmol ^-1 =0,0054757mol

n _NACl =m _NACl /M _NACl =0,350g/58.44g mol ^-1 =0,005989mol

n _{Na2HPO4˙2 H2O} =m _{Na2HPO4˙2 H2O} /M _{Na2HPO4˙2 H2O} =0,077g/177.99g mol ^-1 =0.0004326mol

n _{Na2HPO4˙2 H2O} =m _{Na2HPO4˙2 H2O} /M _{Na2HPO4˙2 H2O} =0,063g/177.99g mol ^-1 =0.0003539mol

n _{Na2HPO4˙2 H2O} =m _{Na2HPO4˙2 H2O} /M _{Na2HPO4˙2 H2O} =0,07g/177.99g mol ^-1 =0.0003932mol

n _{NaH2PO4˙2 H2O} =m _{NaH2PO4˙2 H2O} /M _{NaH2PO4˙2 H2O} =0,0185g/156,01g mol ^-1 =0.0001185mol

n _{NaH2PO4˙2 H2O} =m _{NaH2PO4˙2 H2O} /M _{NaH2PO4˙2 H2O} =0,0145g/156,01g mol ^-1 =0.0000929mol

n _{NaH2PO4˙2 H2O} =m _{NaH2PO4˙2 H2O} /M _{NaH2PO4˙2 H2O} =0,0165g/156,01g mol ^-1 =0.0001057mol

n _H2O =m _H2O /M _H2O =0,17g/18,02g mol ^-1 =0.0094339mol

n _H2O =m _H2O /M _H2O =0,11g/18,02g mol ^-1 =0.0061043mol

n _H2O =m _H2O /M _H2O =0,14g/18,02g mol ^-1 =0.0077691mol

In summary, the unit dose described herein may contain 650 ± 60 mg of 4-amino-7,8-dihydro-L-biopterin and/or 4-amino-(6R,S)-5,6,7 , Free base of 8-tetrahydro-L-biopterin, 140±30mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 16.5±2mg NaH ₂ PO ₄ . 2 H ₂ O, and 350±30mg NaCl. It is also possible that the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin free base, 60±50 mg of crystals Water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 12±2.5mg NaH ₂ PO ₄ ． 2 H ₂ O, and 350±30 mg of sodium chloride (NaCl). In a typical embodiment, this unit dose refers to and is packaged in a 50 ml vial for administration, because an infusion or injection solution with a water-assisted injection of a 50 ml volume of a pharmaceutically acceptable carrier can be administered immediately after reconstitution. Therefore, if, for example, different unit doses are used, such as a 30 ml vial, the amounts of all components of the composition are adjusted accordingly. In the example of a 30ml vial, the amount of each component will be reduced to 3/5 (60%). This means that if the unit dose of a 50ml vial contains 650±60mg of 4-amino-7,8-dihydro-L-biopterin and/or 4-amino-(6R,S)-5,6, 7. Free base of 7,8-tetrahydro-L-biopterin, 140±30mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 16.5±2mg NaH ₂ PO ₄ . 2 H ₂ O, and 350±30mg NaCl, the unit dose of 30ml will contain 390±36mg of 4-amino-7,8-dihydro-L-biopterin and/or 4-amino-(6R, S)-5,6,7,8-Tetrahydro-L-biopterin free base, 84±18mg crystal water, 42±4.2mg Na ₂ HPO ₄ . 2 H ₂ O, 9.9±1.2mg NaH ₂ PO ₄ ． 2 H ₂ O, and 210±18mg NaCl. Accordingly, if the unit dose of a 50 ml vial contains 650 ± 60 mg of 4-amino-7,8-dihydro-L-biopterin and/or 4-amino-(6R,S)-5,6,7 , Free base of 8-tetrahydro-L-biopterin, 60±50mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 12±2.5mg NaH ₂ PO ₄ ． 2 H ₂ O, and 350±30mg NaCl, the unit dose of 30ml will contain 390±36mg of 4-amino-7,8-dihydro-L-biopterin and/or 4-amino-(6R, S)-5,6,7,8-Tetrahydro-L-biological butterfly free base, 36±18mg crystal water, 42±4.2mg Na ₂ HPO ₄ . 2 H ₂ O, 7.2±1.2mg NaH ₂ PO ₄ ． 2 H ₂ O, and 210±18mg NaCl.

The pharmaceutical composition of the present invention may also contain further molecules, for example, related compounds produced in the production process of the pharmaceutical composition of the present invention. Therefore, the pharmaceutical composition of the present invention may contain one, two, three, four, five, six, seven or more additional compounds. Such additional compounds may be selected from the group consisting of, but not limited to, 4-amino-7,8-dihydro-L-biopterin (which may be composed of 4-amino-5,6,7,8- Spontaneous oxidation of tetrahydro-L-biopterin, see above), 4-amino-L-biopterin, (6R,S)-5,6,7,8-tetrahydro-L-biopterin Purine, 1-[(6R,S)-2,4-diamino-5,6,7,8-tetrahydropteridine-6-yl]propanol, 1-[(6R,S)-2, 4-diamino-5,6,7,8-tetrahydropyridin-6-yl]propane, (1R,2S)-1-[(6R,S)-2-(acetylamino)- 4-amino-5,6,7,8-tetrahydropterin-6-yl)-1,2-acetoxy-propane, 2,4-diamino-7,8-dihydropteridine , Or one or more of the compounds in the group consisting of 2,4-diaminopteridine, only a few illustrative compounds are listed.

As outlined previously, pteridine derivatives show properties that are different from other nitric oxide inhibitors, making compounds likely to be more suitable than "typical" arginine analogs. Therefore, the present invention also relates to the use of the freeze-dried pharmaceutical composition of the present invention to treat diseases. In some embodiments, the disease is selected from the group consisting of traumatic brain injury, non-traumatic brain injury, preferably stroke or meningitis, elevated cerebral pressure, and secondary brain injury.

The words "traumatic brain injury" or "brain trauma" occur when the brain is damaged by external force. TBI can be classified according to severity, mechanism (closed or penetrating head injury), or other characteristics (eg, occurring in a specific location or a wide area). Traumatic brain injury can occur due to local head shock, which is the sudden acceleration/deceleration in the skull, or a complex combination of movement and sudden shock, as well as shock waves, or projectile penetration. Glasgow Coma Scale; (GCS), the most commonly used TBI severity classification system, divides the individual's consciousness into 3 to 15 scales, which is based on the response to stimulus speech, movement, and eye opening. Generally speaking, it is agreed that GCS with a TBI of 13 or above is mild, 9 to 12 is moderate, and 8 or below is severe. Similar systems also exist in young children. From a diagnostic point of view, the TBI system between open and closed types is further differentiated. The open TBI system is regarded as an injury, the meninges (dura mater) are mechanically destroyed, and the brain contacts the environment through this opening. Generally, the open TBI system is associated with brain plasma and brain tissue fragments. In closed TBI, the skull or skull is still intact, and the main brain damage (trauma) is characterized by localized lesions, such as contusions or hematomas and/or diffuse cerebral tissue damage. The term "crane" as used herein refers to a collection of nerve skulls (brains) and facial skulls (craniofacial) of bones of vertebrates and cartilage heads. "Intracranial" means located inside the skull.

Conversely, "non-traumatic brain injury" does not involve brain damage caused by mechanical external forces. The causes of non-traumatic brain injury may include lack of oxygen, glucose, or blood. Infection can cause encephalitis (brain edema), meningitis (meningema), or cytotoxicity, for example, due to blastic liver failure and tumors or poisons. These infections can be caused by stroke, heart attack, drowning, suffocation, or diabetic coma, poisoning, or other chemical causes such as alcohol abuse or overdose, infections or tumors, and degenerative conditions such as Alzheimer's and Parkinson's It's happening. The representative of acute neurodegenerative diseases is "stroke", which refers to the loss of brain function. It is due to the disorder of blood supply to the brain. When it occurs rapidly, it is often related to cerebrovascular diseases. This can occur after ischemia (lack of blood flow) caused by obstruction (thrombosis, arterial embolism), or bleeding from the central nervous system (CNS) or intracranial blood vessels. As a result, the affected area of the brain cannot function properly.

Another treatable disease is "meningitis", which is an acute inflammation of the membrane covering the brain and spinal cord, collectively called the meninges. The inflammation may be caused by infection with viruses, bacteria, or other microorganisms, and is less likely to be caused by specific drugs.

In addition to the injury caused by the injury, brain trauma (non-traumatic or traumatic brain injury) causes "secondary injury" or "secondary brain injury", which refers to the occurrence of minutes and days after injury Events. These processes, including changes in intracranial blood flow and intracranial pressure, actually contributed to the initial injury. Secondary injury events may include, for example, injury to the blood-brain barrier, release of inflammatory factors, free radical overload, neurotransmitter glutamate excessive release (excitotoxicity), calcium and sodium ion influx into neurons, and granules Gland disability. After the white matter neurons in the brain are injured, they can detach from their cell bodies due to secondary damage, which may poison their neurons. Other factors of secondary injury are changes in cerebral blood flow; repetitive transient incompleteness of the blood-brain barrier; ischemia (insufficient blood flow); hypoxia in the brain (insufficient oxygen in the brain); cerebral edema (swelling of the brain) ); and increased intracranial pressure (intracranial pressure).

It may also be that intracranial pressure may be increased due to significant effects of edema or lesions (eg bleeding). As a result, cerebral perfusion pressure (cerebral blood flow pressure) decreases and ischemia. When the intracranial pressure rises too high, it can lead to brain death or cerebral hernia, in which part of the brain is compressed by intracranial structures. The term "intracranial pressure (ICP)" refers to the internal pressure of the skull, which is located in the brain tissue and cerebrospinal fluid (CSF). The body has various mechanisms that can maintain ICP stability. The CSF pressure of normal adults changes by about 1 mmHg, which is through CSF Due to changes in production and absorption. The unit of measurement of ICP is millimeters of mercury (mmHg), and the normal value of an adult lying on his back when resting is 7 to 15 mmHg ( http://en.wikipedia.org/wiki/Intracranial_pressure-cite_note-Steiner-1 ). The change in ICP is due to the change in volume of one or more of the chambers contained in the skull. "Elevated cranial pressure" or "increased intracranial pressure" means that the pressure of the skull in an individual is increased compared to a normal, healthy individual. Since intracranial pressure is normally between 7 and 15 mm Hg; therefore, at 20 to 25 mm Hg, the upper limit of the normal value is regarded as an increase in intracranial pressure, and treatment may be required to reduce this pressure. Therefore, the increase of intracranial pressure can be regarded as any pressure of the skull pressure higher than 20mmHg when the individual is supine. , Higher than 30mm Hg, higher than 31mm Hg, higher than 32mm Hg, higher than 33mm Hg, higher than 34mm Hg, or higher than 35mm Hg.

Along this axis, the present invention further relates to the use of the freeze-dried pharmaceutical composition of the present invention for the manufacture of medicaments for the treatment of individuals with intracranial injuries, increased cerebral pressure, and secondary brain injury. As used herein, "intracranial injury" means a type of traumatic brain injury in which the skull and dura mater are still intact (see also above).

The present invention also relates to a method of treating an individual's disease, which includes the step of administering the lyophilized pharmaceutical composition of the present invention to an individual in need.

In terms of administration, the attending physician and clinical factors determine the dosage regimen. As is known in the medical field, the dose of any patient depends on many factors, including the patient's weight, body surface area, age, specific compound to be administered, sex, time, and route of administration, overall health, and simultaneous administration To other drugs. A typical daily dose range can be, for example, 2.5 mg/kg to 12.5 mg/kg body weight; however, doses below or above this exemplary range can also be envisaged, especially considering the above factors.

The dose is preferably given once a day for up to 4 days, however, during the course of treatment, the dose can be given at longer time intervals and, if necessary, at shorter time intervals, such as daily. In the preferred case, the immune response is monitored using the methods described herein and further methods known to those skilled in the art, and the dosage is optimized, such as time, amount, and/or composition. The dosage is variable, but the preferred dose for intravenous administration of Compound (I) and/or Compound (II) is about 2.5 mg/kg to 12.5 mg/kg body weight/day.

If the treatment regimen is injection or continuous short-term infusion, the range should also be approximately 1 μg to approximately 1 mg unit/kg body weight/min. Progress can be monitored by periodic assessment.

In vitro tests can be used arbitrarily to help confirm the optimal dose range. The precise dosage to be used in the formulation also depends on the route of administration and the severity of the disease, and should be based on the judgment of the physician and each The patient's condition is determined. The effective dose can be extrapolated from the dose response curve, which is derived from an in vitro or animal model test system. Preferably, the pharmaceutical composition is administered directly or in combination with an adjuvant.

The maximum daily dose (DDD) is based on the amount of compound (I) and/or (II), which is typically used as the main daily medication indication for an individual (such as an adult). However, the dosage regimen for injection and infusion may be different, or even multiple injections and/or infusions per day. In some embodiments, in the method for treating individual diseases, the maximum daily dose is 12.5 mg/kg body weight, preferably 12.5, 11.5, 10.5, 9.5, 8.5, 7.5, 6.5, 5.5, 4.5, 3.5, 2.5 mg /kg body weight. In some embodiments, the maximum daily dose is 10.0, 9.0, or preferably 8.5 mg/kg body weight, or even lower, such as 7.5, 5.0, 2.5 mg/kg body weight.

The "interval" used in this article is defined as "(from) X to Y", which is equivalent to the interval defined between "X and Y". The two intervals specifically include an upper limit and a lower limit. This means, for example, the interval of "2.5mg/kg to 12.5mg/kg" or "between 2.5mg/kg and 12.5mg/kg", including 2.5, 3.5, 4.5, 5.5, 6.5, 7.5, 8.5, 9.5 , 10.5, 11.5, and 12.5, and any given intermediate value.

It is also envisaged that the pharmaceutical composition is used for co-therapy, that is, co-administered with other agents or drugs, for example, other drugs used to prevent, treat, or ameliorate diseases such as traumatic or non-traumatic brain injury.

The "individual" to be treated by the present invention is preferably a mammal, such as a human, monkey, cat, dog, horse, pig, cow, mouse, or rat, among which humans are preferred.

及/或式(II)化合物：

以及b)至少一磷酸鹽，及任意地NaCl；該方法包含：aa)以緩衝液溶解式(III)及/或(II)化合物：

其中較佳地緩衝液包含磷酸鹽；以及bb)凍乾aa)取得之溶液。 The invention also contemplates a method for preparing a lyophilized solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate, and optionally NaCl; the method comprises: aa) dissolving the compound of formula (III) and/or (II) with a buffer:

Preferably, the buffer solution contains phosphate; and bb) lyophilized solution aa).

Here, it should be noted that although the compound of formula (II) is easily obtained synthetically, a typical synthesis product of the compound of formula (I) is its dihydrochloride dihydrate, as shown in formula (III). For this reason, in preparing the solid group of the present invention containing the compound of formula (I) 4-amino-5,6,7,8-tetrahydro-L-biopterin In the method of the compound, the starting material is typically 4-amino-5,6,7,8-tetrahydro-L-biopterin dihydrochloride dihydrate, that is, the compound of formula (III).

及/或式(II)化合物：

以及b)至少一無機酸，較佳地NaCl；該方法包含：aa)以緩衝液溶解式(III)及/或式(II)化合物：

bb)凍乾aa)取得之溶液。 The invention further relates to a method for preparing a lyophilized solid pharmaceutical composition (suitable for intravenous administration), which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one inorganic acid, preferably NaCl; the method comprises: aa) dissolving the compound of formula (III) and/or formula (II) with a buffer solution:

bb) Lyophilized aa) the solution obtained.

In aa), the buffer is used to dissolve the compound of formula (III) and/or formula (II) to obtain a compound-buffer solution. Generally speaking, a buffer is a solution that contains a weak acid (or base) and a salt of the weak acid (or base). In addition, when a small amount of acid or alkali is added, it resists pH changes. Each phosphate can form several different buffer conjugates. The three possible sodium phosphates may include NaH ₂ PO ₄ -sodium dihydrogen phosphate, Na ₂ HPO ₄ -disodium hydrogen phosphate, and Na ₃ PO ₄ -sodium phosphate.

In aa), the buffer may also be a sodium hydrogen phosphate buffer, which contains at least one phosphate. "Sodium hydrogen phosphate buffer" is a buffer containing sodium phosphate salt. In some embodiments, the sodium phosphate buffer contains at least the sodium monophosphate salt described above. In other specific embodiments, the sodium phosphate buffer contains two, three, four, five, or more sodium phosphate salts, preferably two different phosphates as described above. In another specific embodiment of the method of the present invention, the sodium hydrogen phosphate buffer is prepared by separately dissolving NaH ₂ PO ₄ ˙ ₂ H ₂ O and Na ₂ HPO ₄ ˙ ₂ H ₂ O.

The term "dissolve separately" means that the dissolution of each sodium phosphate occurs separately from each other, as in different glassware. In a further specific embodiment of the method of the present invention, the sodium hydrogen phosphate buffer is pH 7.4, which is obtained by adding NaH ₂ PO ₄ ˙ ₂ H ₂ O solution to Na ₂ HPO ₄ ˙ ₂ H ₂ O solution.

In addition, the buffer of aa) may include NaOH, sodium hydrogen phosphate buffer, and water. Arbitrarily, NaOH is a 5N NaOH solution. "5N NaOH solution" means the equivalent concentration of NaOH (sodium hydroxide) solution is 5 (5N). If the concentration of sodium hydroxide solution is c (NaOH)=5mol/L, the equivalent concentration is 5N.

In some embodiments of the method of the present invention, the buffer solution includes 12-16% (w/w) 5N NaOH, 8-12% (w/w) sodium hydrogen phosphate buffer, and 74-78% (w/w) Water for Injection.

In a further embodiment of the method of the present invention, the pH of the buffer is about 8, 9, 10, 11, 12, 13, or 14.

In yet another specific embodiment of the method of the present invention, the pH of the solution in step aa) is about 4, 5, 6, 7, 8, 9, 10, or 11, preferably between 6.5-7.6, the most Goodland about 7.4.

The buffer can be prepared in different ways. For example, it can be prepared under degassing to obtain degassed buffer. "Degassed buffer" refers to the removal of dissolved gases such as oxygen from liquids (especially water or aqueous solutions). Foaming the solution with an inert gas can replace dissolved harmful and reactive gases, such as oxygen and carbon dioxide. Nitrogen, argon, helium, and other inert gases commonly used. To complete the substitution reaction, the solution should be stirred vigorously and foamed for a long time. In some embodiments, the buffer solution is degassed with nitrogen until the oxygen content is less than 1.0 ppm (parts per million).

In a further additional step of the method of the invention, the solution obtained by aa) can be sterile filtered, preferably with a 0.22 μm filter membrane. The sterile filtration method covers any elimination (removal) or killing of all microbial life forms present on the surface, contained in fluids, medicaments, or compositions (solids or fluids), including infectious agents (such as fungi, bacteria, viruses, Spore form, etc.), the process. Here, sterilization is achieved by filtration. Sterilization can also be achieved by applying heat, chemicals, radiation, high pressure, or a combination thereof, along with the filtration step. The filter membrane can have different pore sizes. Generally, a filter membrane (microfiltration) with a pore size of 0.2 μm will effectively remove microorganisms. When processing biological agents, the virus must be removed or deactivated by heating, glutaraldehyde, or the like. Use a nanofiltration membrane with a pore size of less than 20-50 nm (nanometer filtration). The smaller the pore size, the slower the flow rate. To achieve a higher total flux or to avoid premature clogging, a pre-filtration membrane can be used to protect the small-pore filter membrane.

In another specific embodiment of the method of the present invention, after the pharmaceutical composition is prepared, the lyophilization method is at most 2 hours, preferably at most 1.5 hours, after 1 hour, after 30 minutes, most preferably at most 15 minutes Start later.

It is worth noting that the present invention also relates to the pharmaceutical composition obtained by the method of the present invention.

For better handling of the solid pharmaceutical composition of the present invention, bb) the obtained lyophilisate or the pharmaceutical composition of the present invention is filled into a vial, preferably in an amount of about 1-1.5 g, preferably about 1.25 g of solid formula. Alternatively, bb) the obtained lyophilisate or pharmaceutical composition can be filled into vials, the amount of which is A solid formulation of about 0.9-1.4g, preferably 1.15g. When used as a "unit dose", a solid formulation of about 1-1.5g, or preferably about 1.25g, or a solid formulation of about 0.9-1.4g, preferably 1.15g, is filled into a 50ml vial. The term "vial" refers to a (small) glass or plastic container or bottle, commonly used to hold liquid or solid pharmaceutical compositions. Today, vials are also often made of plastic, such as polypropylene or polystyrene. The solid composition of the present invention can also be stored in any other suitable container.

To obtain a ready-to-use injection, the method of the present invention also includes the step of reconstituting the lyophilisate obtained in bb) into a pharmaceutically acceptable fluid to prepare an injection. The term "injection solution" refers to a solution that can be administered for intravenous injection, preferably for injection or infusion (see also the injections disclosed above). Injections contain pharmaceutically acceptable fluids.

Further features of the invention are listed below:

及/或式(II)化合物：

以及b)至少一磷酸鹽。 Item 1. A solid pharmaceutical composition suitable for intravenous administration, comprising: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate.

Item 2. The pharmaceutical composition of Item 1, wherein the at least one phosphate is sodium phosphate, potassium phosphate, or ammonium phosphate.

Item 3. The pharmaceutical composition according to item 2, wherein the phosphate is selected from the group consisting of Na ₂ HPO ₄ (anhydrous), Na ₂ HPO ₄ ˙2 H ₂ O, Na ₂ HPO ₄ ˙7 H ₂ O, Na ₂ HPO ₄ ˙12 H ₂ O, NaH ₂ PO ₄ (anhydrous), NaH ₂ PO ₄ ˙H ₂ O, NaH ₂ PO ₄ ˙2 H ₂ O, K ₂ HPO ₄ (anhydrous), K ₂ HPO ₄ ˙3 H ₂ O, KH ₂ PO ₄ (anhydrous), and mixtures of them.

Item 4. The pharmaceutical composition of item 2, wherein the phosphate is Na ₂ HPO ₄ ˙ ₂ H ₂ O, and the amount of Na ₂ HPO ₄ ˙ ₂ H ₂ O present in the composition is selected so that The molar ratio of Na ₂ HPO ₄ 2H ₂ O to compound (I) or compound (II) is 0.04 to 0.4.

Item 5. The pharmaceutical composition of item 2, wherein the sodium phosphate is NaH ₂ PO ₄ ˙ ₂ H ₂ O, and the amount of NaH ₂ PO ₄ ˙ ₂ H ₂ O present in the composition is selected so that The range of the molar ratio of NaH ₂ PO ₄ ˙2 H ₂ O to compound (I) or compound (II) is 0.01 to 0.09.

Item 6. The pharmaceutical composition according to any one of Items 1 to 5, which contains two different sodium phosphate salts.

Item 7. The pharmaceutical composition of item 6, wherein the two different sodium phosphate salts are NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O.

Item 8. The pharmaceutical composition as in Item 7, wherein NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O are present in the composition in an amount selected so that NaH ₂ PO ₄ ˙2 H The molar ratio of ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O to compound (I) or compound (ii) ranges from 0.02 to 0.5.

Item 9. The pharmaceutical composition according to any one of Items 1 to 8, wherein Compound (I) and/or Compound (II) are present as a free base.

Item 10. The pharmaceutical composition according to any one of items 1 to 9, wherein the pharmaceutical composition is a lyophilized pharmaceutical composition.

Item 11. The pharmaceutical composition according to any one of items 1 to 10, wherein compound (I) is a compound of formula (Ia):

Item 12. The pharmaceutical composition according to any one of items 1 to 11, wherein compound (II) is a compound of formula (IIa):

Item 13. The pharmaceutical composition according to any one of items 1 to 12, wherein the pharmaceutical composition contains additional pharmaceutical excipients.

Item 14. The pharmaceutical composition of item 13, wherein the additional pharmaceutical excipient is an inorganic salt.

Item 15. The pharmaceutical composition according to Item 14, wherein the inorganic salt is selected from MgCl ₂ , CaCl ₂ , NH ₄ Cl, KCl, or NaCl.

Item 16. The pharmaceutical composition according to Item 15, wherein the inorganic salt is NaCl.

Item 17. The pharmaceutical composition according to Item 16, wherein the amount of NaCl present in the composition is selected so that the molar ratio of NaCl to Compound (I) or Compound (II) is 1.5 to 4, preferably Ground 1.8 to 3.7.

Item 18. The pharmaceutical composition according to any one of Items 1 to 17, wherein the composition further comprises crystal water.

Item 19. The pharmaceutical composition according to any one of items 1 to 18, wherein the composition is suitable for reconstitution in water.

Item 20. The pharmaceutical composition according to any one of items 1 to 19, wherein the composition is suitable for administration by infusion or injection.

Item 21. The pharmaceutical composition according to any one of items 1 to 20, wherein the compound (I) is (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

Item 22. The pharmaceutical composition according to any one of items 1 to 21, wherein the compound (I) is (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

Item 23. The pharmaceutical composition according to any one of items 1 to 22, wherein the compound (I) is a mixture of diastereoisomers, which contains (6R)-4-amino-5,6,7,8 -Tetrahydro-L-biopterin is more than (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin.

Item 24. The pharmaceutical composition according to item 23, wherein (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino-5,6 The amount of ,7,8-tetrahydro-L-biopterin is selected so that (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S) The ratio of the amount of -4-amino-5,6,7,8-tetrahydro-L-biopterin ranges from 0.5 to 2, preferably about 1.3.

Item 25. The pharmaceutical composition according to any one of items 1 to 24, wherein the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro -Free base of L-biopterin, 140±30mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 16.5±2mg NaH ₂ PO ₄ . 2 H ₂ O, and 350±30mg NaCl.

Item 26. The composition of any one of items 1 to 25, wherein the composition comprises one, two, three, four, five, six, seven, or more additional compounds, wherein the additional compounds are selected from A group consisting of one or more of the compounds selected from 4-amino-L-biopterin, (6R,S)-5,6,7,8-tetrahydro-L- Biological butterfly, 1-[(6R,S)-2,4-diamino-5,6,7,8-tetrahydropteridine-6-yl]propanol, 1-[(6R,S)- 2,4-diamino-5,6,7,8-tetrahydropyridin-6-yl]propane, (1R,2S)-1-[(6R,S)-2-(acetylamide )-4-Amino-5,6,7,8-tetrahydropyridin-6-yl)-1,2-acetoxypropane, 2,4-diamino-7,8-dihydro butterfly A group consisting of pyridine and 2,4-diaminopteridine.

Item 27. Use of a lyophilized pharmaceutical composition according to any one of items 1 to 26 to treat diseases.

Item 28. The use according to item 27, wherein the disease is selected from the group consisting of traumatic brain injury, non-traumatic brain injury, preferably stroke or meningitis, elevated cerebral pressure, secondary brain injury.

及/或式(II)化合物：

以及b)至少一磷酸鹽；該方法包含：aa)以緩衝液溶解式(III)及/或(II)化合物：

bb)凍乾aa)取得之溶液。 Item 29. A method for preparing a lyophilized solid pharmaceutical composition suitable for intravenous administration, which comprises: a) a compound of formula (I):

And/or compound of formula (II):

And b) at least one phosphate; the method comprises: aa) dissolving the compound of formula (III) and/or (II) in a buffer solution:

bb) Lyophilized aa) the solution obtained.

Item 30. The method according to item 29, wherein the method further comprises the step of reconstituting the lyophilizate obtained in bb) into a pharmaceutically acceptable fluid to prepare an injection solution.

Item 31. The method according to item 29 or 30, wherein the buffer of aa) is a sodium hydrogen phosphate buffer, which contains at least one phosphate.

Item 32. The method according to any one of items 29 to 31, wherein the buffer of aa) includes NaOH, sodium hydrogen phosphate buffer, and water.

Item 33. The method of item 31, wherein NaOH is a 5N NaOH solution.

Item 34. The method as in item 31, wherein the sodium biphosphate buffer is prepared by dissolving NaH ₂ PO ₄ ˙ ₂ H ₂ O and Na ₂ HPO ₄ ˙ ₂ H ₂ O, respectively.

Item 35. The method according to item 33, wherein the sodium hydrogen phosphate buffer is pH 7.4, which is obtained by adding NaH ₂ PO ₄ ˙ ₂ H ₂ O solution to Na ₂ HPO ₄ ˙ ₂ H ₂ O solution.

Item 36. The method according to any one of items 29 to 35, wherein the buffer contains 12-16% (w/w) 5N NaOH, 8-12% (w/w) sodium hydrogen phosphate buffer, and 74-78 %(w/w) water for injection.

Item 37. The method according to any one of items 29 to 36, wherein the solution obtained by aa) is sterile filtered, preferably using a 0.22 μm filter membrane.

Item 38. The method of any one of items 29 to 37, wherein the pH of the buffer is about 8, 9, 10, 11, 12, 13, or 14.

Item 39. The method according to any one of items 29 to 38, wherein the pH of the solution of step aa) is about 4, 5, 6, 7, 8, 9, 10, or 11, preferably between 6.5 and Between 7.6, optimally 7.4.

Item 40. The method according to any one of items 29 to 39, wherein the lyophilisate obtained in bb) is filled into a vial, preferably in an amount of about 1-1.5 g, preferably 1.25 g of solid formulation.

Item 41. The method of any one of items 29 to 40, wherein the buffer is prepared with degassed buffer.

Item 42. The method of any one of items 29 to 41, wherein the buffer is degassed with nitrogen until the oxygen content is less than 1.0 ppm.

Item 43. The method according to any one of items 29 to 42, wherein after the solution is prepared, the lyophilization method starts at most 2 hours.

Item 44. A pharmaceutical composition obtained by the method as in any one of items 29 to 43.

Item 45. The use of the lyophilized pharmaceutical composition according to any one of items 1 to 28 to prepare a medicament for the treatment of individuals with intracranial injuries, elevated cerebral pressure, and secondary brain injury.

Item 46. A method of treating an individual's disease, which includes the step of administering a lyophilized pharmaceutical composition according to any one of items 1 to 28 to an individual in need.

Item 47. The method of item 46, wherein the maximum daily dose is 20 mg/kg body weight, preferably, 17.5, 15.0 or 12.5, 10, 8.5, 7.5, 5.0, 2.5 mg/kg body weight.

Examples

The following examples further illustrate the invention. However, these embodiments should not be construed as limiting the scope of the present invention. The covered embodiments are intended to illustrate, and the present invention is limited only by the scope of patent application.

Example 1-Manufacturing process description

VAS203 is prepared in a multi-step synthesis, which starts with the commercially available L-biopterin. Figure 1 shows the flow chart. The synthesis of VAS203 is based on the literature by W. Pfleiderer et al. in Pteridines 1989, 1, 199-210 and Pteridines 1995, 6, 1-7.

The purpose of the first stage of development is to reproduce the documentation process and assess the feasibility of its multi-gram scale. Exclude intermediate purification by flash chromatography and replace with precipitation or crystallization steps. In addition, the molar ratio of the collected reagents and the reaction conditions were carried out so as to achieve a sufficiently large intermediate MAE 119 (4-amino-L-biopterin) treatment capacity.

PtO2 is used as a catalyst to establish a sufficiently large and reliable way to hydrogenate MAE 119 to VAS203. Test aqueous media with different solvents and different pH values, but only when diluted hydrochloric acid is used to achieve fast and reproducible results, especially in the ratio of diastereomers of drug substances. The total residual platinum load can be greatly reduced by the prehydrogenation of the platinum(IV) oxide catalyst, so that the ratio of diastereomers slightly changes. Post-treatment of VAS203 with an acidic solution proves to be difficult, because the simple evaporation of water will leave a glassy residue under reduced pressure. When 2-propanol is used as the co-solvent in the evaporation process, a breakthrough is possible. This procedure produced a solid, but unfortunately, 2-propanol penetrated the solid. With conventional drying conditions, the amount of residual 2-propanol cannot be reduced to less than 5% (w/w). To overcome this problem, a final freeze-drying step can be included to produce VAS203. The individual batches can be pooled to obtain larger quantities.

Evidence of structural assignment of VAS203 was provided by a synthetic route, which was supported by elemental analysis, nuclear magnetic resonance ( ¹ H-NMR and ¹³ C-NMR), ultraviolet, and infrared spectroscopy (data not shown).

Example 2

Description and composition of drug products

VAS203 will be supplied as sterile, white to light red or brown lyophilized powder, which is a 50mL glass vial filled with nitrogen to protect the environment. Each vial contains 650±60mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin free base and 140±30mg of crystal water. In addition, a vial containing 350 ± 30mg of sodium chloride (NaCI), sodium phosphate dibasic dihydrate of 70 ± 7mg _{(Na 2 HPO 4 .2 H 2} O), and 16.5 ± 2mg of sodium dihydrogen phosphate dihydrate (NaH ₂ PO _4. 2 H ₂ O). The tolerance range of the drug product composition is quite high (±10%). The reason is that the hydrochloric acid content of VAS203 changes. The hydrochloric acid content of VAS203 varies from batch to batch, up to 10% (2.03 HCl to 2.24 HCl). During the preparation of the drug product, the hydrochloric acid of the present invention is neutralized by adding sodium hydroxide and sodium phosphate buffer to obtain an isotonic solution with a physiological pH. Therefore, the molecular contents (sodium chloride, disodium hydrogen phosphate, and sodium dihydrogen phosphate) generated during neutralization vary according to the hydrochloric acid content of each batch of VAS203. The given tolerance range is required to meet the specifications of quality-related parameters pH and osmotic pressure. The qualitative composition of one gram of VAS203 vials is shown in Table 2 .

Pharmaceutical development

This article is to develop a solid lyophilized dosage form of VAS203 to prepare an infusion solution.

Add 1g of VAS 203 and 10g of sodium hydroxide to the sodium hydrogen phosphate solution buffer to make the final pH 7.4, and take it for aseptic processing, membrane filtration and sterilization, and fill to a 50mL glass vial. Subsequently, according to the selected lyophilization procedure, the solution was freeze-dried to produce a freeze-dried product with excellent stability. In this solid composition, VAS 203 is present as the free base of 4-amino-5,6,7,8-tetrahydro-L-biopterin. The vial was closed under nitrogen, sealed with a freeze-dried stopper, and closed with a white vacuum bottle cap. To provide an isotonic solution, an excipient is added, which has a physiological pH after being reconstituted with 50 mL of water for injection. The pH of the final isotonic solution is 6.5 to 7.6. The final concentration of the drug substance VAS203 is 20 mg/mL.

Description of manufacturing process and process control

At pH 7.4, 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin dihydrochloride dihydrate (VAS203) dissolved in solution buffer is not Stable and oxidized to its two metabolites 4-amino-7,8-dihydro-L-biopterin and 4-amino-L-biopterin. Therefore, the important point is that the VAS203 solution should be prepared with degassed buffer, and after the VAS203 solution is prepared, the lyophilization method should be started without delay.

To lyophilize a 50mL vial, a concentrated VAS203 solution buffer (1g VAS203 plus 10g of buffer) must be prepared. Prepare a buffer solution by mixing stock solutions:

14% (w/w) 5N sodium hydroxide (NaOH) solution

10% (w/w) sodium hydrogen phosphate buffer (NaPB) solution

500mmol/L, pH 7.4

76% (w/w) water for injection

The sterile solution buffer is then degassed with nitrogen until the oxygen content is less than 1.0 ppm. For each vial, 1.0 g of VAS203 was dissolved in 9.0 g of this solution buffer. The solution buffer was filled into a bottle flushed with nitrogen, and the solid (lyophilized) VAS203 was added to it carefully within 15 minutes under a nitrogen protective environment. After checking the pH value, the VAS203 solution was sterile filtered with a 0.22 μm Millipak 200 filter membrane. After use, the integrity of the 0.22 μm filter membrane was tested. Take a 100mL sample to test the bioburden. The bioburden limit value of the drug product solution before filtration and sterilization is defined as 10cfu/100mL. The residual solution was filtered a second time and dispensed into 50 mL vials, 10 g per vial. The vial is sterilized before dry heat filling, according to section 5.1.1. of the European Pharmacopoeia. After the freeze-drying method, the vial was closed under nitrogen, sealed with a freeze-dried stopper, and closed with a white vacuum bottle cap. VAS203 solution was prepared quickly before lyophilization. Verify the aseptic filling process with media fill. The flow chart of the feasible manufacturing process steps and the components used in each step are shown in Figure 2 . The result is a solid composition in which VAS203 is present as the free base of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L-biopterin.

Stability

The stability monitoring system is conducted for 24 months. The samples are stored "long-term" at 5°C (verum). The accelerated stability study was conducted at 40°C/75% relative humidity for 6 months (verum). Analytical procedures used for stability procedures include testing, purity, and testing of related substances. For the release of drug products, the application specifications and methods of use are the same.

The stability monitoring department is conducted for at least 60 months. Table 3 shows the conditions and storage time of the stability investigation. The samples are stored "long-term" at 5°C (verum). The accelerated stability study was conducted at 40°C/75% relative humidity for 6 months (verum). Table 4 provides test schedules at different time points. Analytical procedures used for stability procedures include testing, purity, and testing of related substances. to For drug product release, the application specifications and methods of use are the same. Table 5 shows exemplary results of stability studies (which are illustrated in Figure 4).

discuss

The stability data is taken from the batch stored at 5°C and 40°C/75% relative humidity, which is carried out in the closed light. Samples stored at two temperatures for up to 24 months observed slight changes, but they showed no consistent trend and are believed to fall within the range of analytical variability. No degradation products were formed. The first metabolite (oxidation product) of the drug substance (4-amino-7,8-dihydro-L-biopterin) can be observed To a slight increase. After 24 months of storage at 5°C, 7 out of 10 vials found a total of 15 visible particles.

The stability data is taken from the batch stored at 5°C and 40°C/75% relative humidity, which is carried out in the closed light. No significant changes were detected in samples stored at two temperatures for up to 36 months. No degradation products were formed.

in conclusion

When stored in the closed light at 5 ℃, the stability of the VAS203 drug product vial is not less than 36 months. When stored in the closed light at 40℃/75% relative humidity, the stability of the vial of VAS203 drug product is not less than 6 months. Considering the above, the shelf life of 1g of filled lyophilized VAS203 powder in a 50mL glass vial under a nitrogen protective environment is 42 months. The storage instructions will recommend vials stored at 2-8°C, and the vials must be wrapped in aluminum foil and closed in a carton. All reconstituted drug products should be visually inspected for specific substances. The storage instructions will recommend storing vials at 2-8°C.

If the stability study of 1g VAS203 vial and 50mL placebo vial can be carried out at the same time to obtain appropriate long-term and accelerated stability data, the shelf life can be extended, and the results meet the existing specifications. The shelf life is based on the principle described in the ICH Q1E guidelines: if no significant changes are detected within 6 months under accelerated conditions, and long-term data shows little or no change over time and low variability, the actual time data will be used. Double the time as the extended warranty period, but the warranty period should not be greater than 12 months of the length of time that long-term data can be obtained.

Example 3-Stability of administration preparation and infusion process

A stability study of the drug substance VAS203 at room temperature was conducted to test any degradation of the recombinant solution in a transparent 50 mL polypropylene (PP) syringe, which was used in clinical research. During the stability test, the PP syringe was exposed to normal sunlight. Solution contents, purity, related substances, and pH were monitored for 48 hours (after preparation, starting, 6, 24, 48 hours). Table 6 shows the stability test results.

discuss

Within 48 hours, the content (test) of the solution within the specified range (650±60 mg) remained unchanged. During the test period, the chromatographic purity of the two non-image isomers fell within the specified range, and no obvious changes were observed. Relevant contents of related substances remained within the specified range within 48 hours. No degradation products formed. A slight increase in the first metabolite of VAS203 (4-amino-7,8-dihydro-L-biopterin) can be observed. However, this oxidation may be caused by oxygen remaining in the syringe, and the change is not obvious between 6 and 48 hours. The pH of the recombinant VAS203 solution remained within the specified range within 48 hours. No change was observed. No photosensitivity was observed for up to 48 hours.

in conclusion

The investigation conducted confirmed that the VAS203 reconstitution solution was stable in its application device (50 mL PP syringe) when exposed to normal sunlight for 48 hours at room temperature. However, use-by date should be reduced to avoid microbial contamination. "Daily use" is defined as the time of preparation of the VAS203 solution plus 27 hours.

Table 6. Stability of VAS203 recombinant solution in 50mL PP syringe (applied device) when stored at room temperature

As used herein, it should be noted that unless otherwise specified, the singular forms "a", "one", and "the" include plural forms. Thus, for example, reference to "a reagent" includes one or more such different reagents, and reference to "the method" includes reference to equivalent steps and methods known to those of ordinary skill in the art, which can be improved by the methods described herein Or replace.

All publications and patents cited by the present invention are hereby incorporated into this case as reference materials. To the extent that the incorporated reference materials are inconsistent or inconsistent with this specification, this specification will be used as the basis.

Unless otherwise specified, the word "at least" before a series of elements is understood to mean each element of the series. Those skilled in the art should understand or be able to use experiments that do not exceed routines, many of which are equivalent to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the present invention.

Throughout this specification and the scope of patent applications disclosed, unless the full text requires otherwise, the word "include" and its similar words such as "include" and "include" will be understood to imply the inclusion of the stated integers or steps, Or groups of integers or steps, and does not exclude any other integers or steps, or groups of integers or steps. As used herein, the word "contained" may be replaced by the word "contained", or sometimes used in this article with "to".

As used herein, "consisting of" excludes any elements, steps, or ingredients not specified in the scope of the patent application. As used herein, "basically composed of" does not exclude materials or steps that do not substantially affect the basis and novel features of the patent scope of this application. In each case of this article, any of the words "including", "basically composed of ~", and "consisting of ~" may be replaced by one of the other two words.

Several documents are cited throughout this specification. Each of the documents cited here (including all patents, patent applications, scientific publications, manufacturer's instructions, instructions for use, etc.), whether above or below, is hereby incorporated by reference in this case. This article cannot be interpreted as agreeing that the present invention has no right to be disclosed prior to the previous invention.

Claims (32)

A solid pharmaceutical composition comprising: a) a compound of formula (I):

And/or compound of formula (II):

And b) two different sodium phosphate salts, wherein the two different sodium phosphate salts are NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O, and among them NaH ₂ PO ₄ ˙2 H ₂ O The amount of Na ₂ HPO ₄ ˙2 H ₂ O present in the composition is selected so that both NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O are related to compound (I) or compound (ii) The molar ratio ranges from 0.02 to 0.5. The pharmaceutical composition according to claim 1, wherein the compound (I) and/or the compound (II) is present as a free base. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is a freeze-dried pharmaceutical composition. The pharmaceutical composition according to claim 1, wherein compound (I) is a compound of formula (Ia):

The pharmaceutical composition according to claim 1, wherein compound (II) is a compound of formula (IIa):

The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises additional pharmaceutical excipients. The pharmaceutical composition according to claim 6, wherein the additional pharmaceutical excipient is an inorganic salt. The pharmaceutical composition according to claim 7, wherein the inorganic salt is selected from MgCl ₂ , CaCl ₂ , NH ₄ Cl, KCl, or NaCl. The pharmaceutical composition according to claim 8, wherein the inorganic salt is NaCl. The pharmaceutical composition according to claim 9, wherein the amount of NaCl present in the composition is selected so that the molar ratio of NaCl to compound (I) or compound (II) ranges from 1.5 to 4. The pharmaceutical composition according to any one of claims 1 to 10, wherein the composition further comprises crystal water. The pharmaceutical composition according to any one of claims 1 to 4, wherein the compound (I) is (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin. The pharmaceutical composition according to any one of claims 1 to 4, wherein the compound (I) is (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin. The pharmaceutical composition according to any one of claims 1 to 4, wherein the compound (I) is a mixture of diastereomers, which contains (6R)-4-amino-5,6,7,8-tetra There are more hydrogen-L-biopterin than (6S)-4-amino-5,6,7,8-tetrahydro-L-biopterin. The pharmaceutical composition according to claim 14, wherein (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4-amino-5,6,7 The amount of ,8-tetrahydro-L-biopterin is selected so that (6R)-4-amino-5,6,7,8-tetrahydro-L-biopterin and (6S)-4 -The ratio of the amount of amino-5,6,7,8-tetrahydro-L-biopterin ranges from 1.1 to 2. The pharmaceutical composition according to any one of claims 1 to 4, wherein the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L -Free base of biological pterin, 140±30mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 16.5±2mg NaH ₂ PO ₄ . 2 H ₂ O, and 350±30mg NaCl. The pharmaceutical composition according to any one of claims 1 to 4, wherein the unit dose of the composition contains 650±60 mg of 4-amino-(6R,S)-5,6,7,8-tetrahydro-L -Free base of Pteropterin, 60±50mg crystal water, 70±7mg Na ₂ HPO ₄ . 2 H ₂ O, 12±2.5mg NaH ₂ PO ₄ ． 2 H ₂ O, and 350±30mg NaCl. The pharmaceutical composition according to any one of claims 1 to 4, wherein the composition comprises one, two, three, four, five, six, seven or more additional compounds, wherein the additional compounds are selected from A group consisting of one or more of the compounds selected from 4-amino-L-biopterin, (6R,S)-5,6,7,8-tetrahydro-L-biological Pterin, 1-[(6R,S)-2,4-diamino-5,6,7,8-tetrahydropteridine-6-yl]propanol, 1-[(6R,S)-2 ,4-diamino-5,6,7,8-tetrahydropyridin-6-yl]propane, (1R,2S)-1-[(6R,S)-2-(acetylamino) -4-amino-5,6,7,8-tetrahydropterin-6- Group]-1,2-acetoxypropane, 2,4-diamino-7,8-dihydropteridine, 2,4-diaminopteridine group. A method for preparing a lyophilized solid pharmaceutical composition, comprising: a) a compound of formula (I):

And/or compound of formula (II):

And b) two different sodium phosphate salts, wherein the two different sodium phosphate salts are NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O, and among them NaH ₂ PO ₄ ˙2 H ₂ O The amount of Na ₂ HPO ₄ ˙ ₂ H ₂ O present in the composition is selected so that NaH ₂ PO ₄ ˙ ₂ H ₂ O and Na ₂ HPO ₄ ˙ ₂ H ₂ O and compound (I) or compound (ii ) The molar ratio ranges from 0.02 to 0.5, and optionally NaCl; the method includes: aa) dissolving the compound of formula (III) and/or (II) in a buffer solution:

Wherein the buffer contains the two different sodium phosphate phosphates; bb) lyophilized aa) the obtained solution. A method for preparing an injection, comprising: a) a compound of formula (I):

And/or compound of formula (II):

And b) two different sodium phosphate salts, wherein the two different sodium phosphate salts are NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O, and among them NaH ₂ PO ₄ ˙2 H ₂ O The amount of Na ₂ HPO ₄ ˙2 H ₂ O present in the composition is selected so that both NaH ₂ PO ₄ ˙2 H ₂ O and Na ₂ HPO ₄ ˙2 H ₂ O are related to compound (I) or compound (ii) The molar ratio ranges from 0.02 to 0.5, and optionally NaCl; the method includes: aa) dissolving the compound of formula (III) and/or (II) with a buffer solution:

Wherein the buffer contains the two different sodium phosphate phosphates; bb) lyophilized aa) the obtained solution; cc) the lyophilized substance obtained from bb) is reconstituted into a pharmaceutically acceptable fluid to prepare an injection solution, wherein the bb ) The obtained lyophilisate is filled into vials. The method according to claim 20, wherein the lyophilisate obtained in bb) is filled into a 50ml vial, (i) a solid formulation with an amount of 1-1.5g; or (ii) a solid formulation with an amount of 0.9-1.4g. The method of claim 19 or 20, wherein NaOH is a 5N NaOH solution. The method of claim 22, wherein the sodium biphosphate buffer is prepared by dissolving NaH ₂ PO ₄ ˙ ₂ H ₂ O and Na ₂ HPO ₄ ˙ ₂ H ₂ O, respectively. The method according to claim 23, wherein the sodium hydrogen phosphate buffer is pH 7.4, which is obtained by adding NaH ₂ PO ₄ ˙ ₂ H ₂ O solution to Na ₂ HPO ₄ ˙ ₂ H ₂ O solution. The method according to any one of claims 19 to 21, wherein the buffer contains 12-16% (w/w) 5N NaOH, 8-12% (w/w) sodium hydrogen phosphate buffer, and 74-78% (w/w) Water for injection. The method according to any one of claims 19 to 21, wherein the solution obtained by aa) is sterile filtered. The method according to any one of claims 19 to 21, wherein the pH of the buffer is 8, 9, 10, 11, 12, 13, or 14. The method according to any one of claims 19 to 21, wherein the pH value of the solution of step aa) is 4, 5, 6, 7, 8, 9, 10, or 11. The method according to any one of claims 19 to 21, wherein the buffer is prepared with a degassed buffer. The method of any one of claims 19 to 21, wherein the buffer is degassed with nitrogen until the oxygen content is less than 1.0 ppm. The method according to any one of claims 19 to 21, wherein after the solution is prepared, the lyophilization method is started at most 2 hours later. Use of the pharmaceutical composition according to any one of claims 1 to 18 to prepare a medicament for the treatment of individuals with intracranial injuries, increased cerebral pressure, and secondary brain injury.

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