CN113214337B

CN113214337B - Pharmaceutical co-crystal of progesterone, pyridine and derivatives thereof

Info

Publication number: CN113214337B
Application number: CN202110545899.5A
Authority: CN
Inventors: 徐娟; 宁丽峰; 马进
Original assignee: Institute Of Science And Technology National Health Commission
Current assignee: Institute Of Science And Technology National Health Commission
Priority date: 2021-05-19
Filing date: 2021-05-19
Publication date: 2022-01-11
Anticipated expiration: 2041-05-19
Also published as: CN113214337A

Abstract

The invention discloses a pharmaceutical co-crystal of progesterone, pyridine and derivatives thereof, wherein the molecular ratio of the progesterone to the co-formed substance is 1: 1. The invention also discloses a preparation method and application of the pharmaceutical co-crystal of progesterone, pyridine and derivatives thereof.

Description

Pharmaceutical co-crystal of progesterone, pyridine and derivatives thereof

Technical Field

The invention belongs to the field of biological medicines, and relates to a pharmaceutical cocrystal of progesterone, pyridine and derivatives thereof.

Background

Progesterone (pregestrone), also known as progesterone, 4-pregnane-3, 20 dione, is a progestin that is secreted by the female ovarian corpus luteum. Researchers successfully synthesize exogenous progesterone for the first time in the 30 th century, and play an important role in the female menstrual period and the female pregnancy (written by the organization of the reasonant expert committee of the Ministry of health, Chinese physicians, clinical medication guide [ M ] Chongqing Press 2014.).

The female ovary of natural conception can generate corpus luteum after ovulation, gonadotropin generated by a human body can correspondingly stimulate the corpus luteum to secrete progesterone after embryo implantation, the main physiological effect of the progesterone is to recognize pregnancy of animals and maintain pregnancy, the acting parts are mostly on uterine muscles and endometrium, and the progesterone has auxiliary effect on implantation of fertilized eggs and is necessary hormone for maintaining pregnancy. Progesterone acts in conjunction with estrogen to regulate various changes in the reproductive phase of the body.

The clinical application of progesterone mainly comprises the following aspects: (1) threatened abortion: the disease is caused by various factors, including embryonic factors, environmental factors, maternal factors and the like, and luteal insufficiency of the body is one of the main reasons, so that the placenta is not developed enough and the fetus is not developed enough and stable. The progesterone can effectively improve the hormone level, improve the physiological environment of the body, facilitate pregnancy, improve the pregnancy rate of women, provide a better environment for the development of embryos and protect the development of the embryos. (2) Premenstrual tension syndrome: premenstrual tension Syndrome (PMS) refers to a state of weakness of a female body in the menstrual cycle, and compared with daily life, the PMS is anxious, easy to fatigue, easy to insomnia and the like, and frequently occurs in the later period of the menstrual cycle. The reduction of progesterone secretion in the later phases of the menstrual cycle is the main cause of PMS, and the reduction of hormone levels leads to a reduction in the content of tetrahydroprogesterone, a decomposition product thereof, and a reduction in hormones exerting a sedative and analgesic effect in the body causes many maladaptive symptoms in the body. The progesterone is administered for hormone therapy, so that symptoms of menstrual tension syndrome can be effectively relieved, and the health of the body can be maintained. (3) Anovulatory dysfunctional uterine bleeding and amenorrhea: anovulatory dysfunctional uterine bleeding is a type of Dysfunctional Uterine Bleeding (DUB), and about 80% of affected people are adolescent or menopausal women (Liuwei. gynecological Yangrong capsule combined with norethindrone for treating anovulatory dysfunctional uterine bleeding with curative effect observation [ J ]. Shanxi Chinese medicine, 2015,36(4): 467-8.). The main pathogenic causes are the disturbance of secretion system, the disturbance of the regulation mechanism of hypothalamus-pituitary-ovary and the irregular bleeding of uterus, if the treatment can not be carried out in time, the secondary anemia, infection and infertility of serious patients can be caused. The pathogenic cause of amenorrhea is the same as that of anovulatory dysfunctional uterine bleeding. The two diseases are common gynecological diseases, are not common, and are often accompanied with symptoms of abdominal pain, abdominal distension, breast distending pain and the like. The progesterone capsule auxiliary medicine estradiol valerate is added for treating anovulatory dysfunctional uterine bleeding and amenorrhea, the thickness of endometrium can be reduced, the menstrual cycle and menstrual period can be effectively shortened, a better treatment effect is obtained, and the safety is higher. (4) Auxiliary reproduction technology: assisted Reproductive Technology (ART) refers to pregnancy of sterile couples by medical assistance, and includes two major categories of Artificial Insemination (AI) and In vitro fertilization-Embryo Transfer (IVF-ET) and derivatives thereof; luteal phase support therapy is an indispensable procedure in vitro fertilization, and the use of progesterone can obviously improve the function of the luteal phase, so that embryos can be smoothly planted, and the success rate of ART is effectively improved. (5) Protective action on the nervous system: progesterone can protect the nervous system and promote its regeneration through a variety of action musculature (Robertson C L, Fidan E, Stanley R M, et al. Progesterone for Neuroprotection in peripheral Traumatic Brain in J. peripheral cardiac Medicine: a journal of the Society of clinical Care Medicine and the World Federation of peripheral Intensive and clinical Care Society, 2015,16(3): 236-44; Singh M, Suc. Progesterone, BDNF and Neuroprotection [ J ] Neuroscience,2013,239: 84.). (6) Cardiovascular protection: the progesterone can inhibit the expression of the vascular cell adhesion factor (VCAM-1), reduce the adhesion among cells, effectively protect arterial blood vessels and avoid the occurrence of AS. (7) Treatment of respiratory diseases: progesterone can regulate the respiratory function of the body by directly improving the self-function of pulmonary alveoli. The progesterone is used for periodic treatment of menstrual asthma, and can directly relieve bronchospasm of patients, enhance the activity of agonist, thereby improving the relief of asthma, relieving edema and eliminating dysnoesia.

Progesterone is poor in solubility, almost insoluble in water, and only 2mg/L in solubility, and is usually administered by injection of progesterone oil injection in clinic. Its advantages are sure curative effect, low cost and the following disadvantages: the pain and stimulation of the injection part are easy to form local induration, and the absorption and recovery of the local induration and the aseptic abscess need a long time.

The development of pharmaceutical crystalline forms is one possibility to increase bioavailability and, in addition to bioavailability, may improve several other properties, such as dissolution, physical stability, mechanical properties, hygroscopicity, chemical stability, flowability and purification processing ability. Therefore, the research of new crystal forms is an important means for drug development.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a combined pharmaceutical co-crystal formed by progesterone, pyridine and derivatives thereof and application thereof, aiming at the problems that the existing progesterone is poor in solubility and the drug effect cannot be well exerted.

In order to solve the technical problems, the invention adopts the following technical scheme:

a first aspect of the invention provides a co-crystal of progesterone and a co-form selected from pyridine and/or derivatives thereof.

Further, progesterone and the conformer are present in the co-crystal in a 1:1 molecular ratio.

Further, the pyridine and/or its derivatives are selected from: 2, 6-diaminopyridine, 4-pyridinecarboxamide, 4-aminopyridine.

Further, the experimental formula of the progesterone-2, 6-diaminopyridine eutectic is C₂₆H₃₇N₃O₂。

The experimental formula of the progesterone-4-pyridine carboxamide eutectic is C₂₇H₃₆N₂O₃；

The experimental formula of the progesterone-4-aminopyridine eutectic is C₂₆H₃₆N₂O₂。

According to the co-crystal of the first aspect of the invention, the co-crystal has an X-ray spectrum with characteristic 2 θ values at 2 θ ± 0.2 ° respectively:

progesterone-2, 6-diaminopyridine co-crystal: 13.402, 16.92, 19.279, 19.957, 25.241;

progesterone-4-pyridinecarboxamide co-crystal: 13.5, 20.421;

progesterone-4-aminopyridine co-crystal: 13.717, 15.899, 16.079, 21.441.

Further, the co-crystal also has an X-ray spectrum with characteristic 2 θ values at 2 θ ± 0.2 ° below, respectively:

progesterone-2, 6-diaminopyridine co-crystal: 10.459, 14.799, 15.078, 18.679, 23.458, 23.879;

progesterone-4-pyridinecarboxamide co-crystal: 13.1, 15.022, 17.401, 18.282, 22.218;

progesterone-4-aminopyridine co-crystal: 14.3, 15.499, 18.499, 19.321, 19.92, 26.041, 29.64.

progesterone-2, 6-diaminopyridine co-crystal: 8.539, 15.693, 20.919, 21.139, 23.138, 26.96, 29.737, 31.602;

progesterone-4-pyridinecarboxamide co-crystal: 21.92, 26.418, 27.516;

progesterone-4-aminopyridine co-crystal: 17.382, 22.619, 25.599, 28.721.

Further, the X-ray powder diffraction pattern is substantially as shown in fig. 6.

According to the co-crystal of the first aspect of the present invention, the co-crystal comprises endothermic peaks at about the following melting point temperatures, respectively:

progesterone-2, 6-diaminopyridine co-crystal: 111.1 ℃;

progesterone-4-pyridinecarboxamide co-crystal: 140.4 ℃;

progesterone-4-aminopyridine co-crystal: 127.4 ℃.

Further, the thermogram is substantially as shown in fig. 1.

According to the eutectic of the first aspect of the present invention, the eutectic respectively comprises the following crystal parameters:

the progesterone-2, 6-diaminopyridine eutectic is monoclinic system, P2₁Space group, cell parameter of

α ═ γ ═ 90.00 °, β ═ 107.375(4) °, Z ═ 8, and the unit cell volume is

The progesterone-4-pyridine carboxamide eutectic is orthorhombic, P2 ₁2₁2₁Space group, cell parameter of

α ═ β ═ γ ═ 90.00 °, Z ═ 8, and unit cell volume

The progesterone-4-aminopyridine eutectic is orthorhombic, P2 ₁2₁2₁Space group, cell parameter of

α ═ β ═ γ ═ 90.00 °, Z ═ 4, and unit cell volume

A second aspect of the present invention provides a method for producing a eutectic crystal according to the first aspect of the present invention, including:

1) the progesterone and the copolymer are crushed to obtain a white powder sample,

2) and adding the white powder sample into an ester solvent for dissolving, and volatilizing to obtain colorless blocky crystals.

Further, the solvent is ethyl acetate.

Further, the molecular ratio of progesterone to co-form is 1: 1.

Further, progesterone and the co-form were pulverized using a ball mill.

In a third aspect, the present invention provides a pharmaceutical composition comprising a co-crystal according to the first aspect of the present invention.

Further, the pharmaceutical composition also comprises a pharmaceutically acceptable carrier.

Further, the administration mode of the pharmaceutical composition comprises oral administration, injection, vaginal administration and transdermal administration.

In a fourth aspect, the invention provides the use of a co-crystal according to the first aspect of the invention or a pharmaceutical composition according to the third aspect of the invention in the manufacture of a medicament for maintaining pregnancy.

In a fifth aspect, the present invention provides the use of a co-crystal according to the first aspect of the present invention or a pharmaceutical composition according to the third aspect of the present invention in the manufacture of a medicament for the prevention and/or treatment of gynaecological disorders.

Further, the gynecological diseases include spontaneous premature birth, corpus luteum insufficiency, secondary amenorrhea, premenstrual syndrome.

According to a sixth aspect of the present invention, there is provided a use of the co-crystal according to the first aspect of the present invention or the pharmaceutical composition according to the third aspect of the present invention for the preparation of a medicament for the prevention and/or treatment of cardiovascular diseases and nervous system diseases.

In the present invention, the relative amounts of the active ingredient, pharmaceutically acceptable carrier and/or any additional ingredients in the pharmaceutical composition will vary depending on the identity, size and/or condition of the subject being treated and further depending on the route of administration of the composition. The composition may comprise from 0.1% to 100% (w/w) of the active ingredient.

Pharmaceutically acceptable carriers used in preparing the provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surfactants and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents and/or oils. Excipients such as cocoa butter and suppository waxes, colorants, coatings, sweeteners, flavoring agents and flavoring agents may also be present in the composition.

Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active agent, the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1, 3-butylene glycol, ethylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, oral compositions can contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In some embodiments for parenteral administration, the complexes of the invention are mixed with solubilizing agents such as alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and combinations thereof.

Injectable preparations, for example, sterile aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable formulation may be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable vehicles and solvents that can be used are water, ringer's solution, u.s.p., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other injectable sterile medium prior to use.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or: a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders, for example carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia, c) humectants, such as glycerol, d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate, e) solution retarding agents, such as paraffin, f) absorption promoters, such as quaternary ammonium compounds, g) wetting agents, such as cetyl monostearate and glycerol monostearate, h) absorbents, such as kaolin and bentonite, and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents.

Solid compositions of a similar type may be employed as fillers in soft and hard-filled capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared using coatings and shells such as enteric coatings and other coatings well known in the pharmacological arts. They may optionally contain opacifying agents and may be of a composition that they release the active ingredient or ingredients in only, or preferably, a part of the intestinal tract, optionally, in a delayed release manner. Examples of encapsulation compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may be employed as fillers in soft and hard-filled capsules using excipients such as lactose, as well as high molecular weight polyethylene glycols and the like.

The active ingredient may be in microencapsulated form with one or more of the excipients mentioned above. The solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings, controlled release coatings and other coatings well known in the art of pharmaceutical formulation. In such solid dosage forms, the active ingredient may be mixed with at least one inert diluent (e.g., sucrose, lactose or starch). Such dosage forms may contain, as is common practice, other substances in addition to inert diluents, e.g., tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, the dosage forms may contain buffering agents. They may optionally comprise opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferably, in a specific part of the intestinal tract, optionally, in a delayed manner. Examples of encapsulants that can be used include, but are not limited to, polymeric substances and waxes.

Dosage forms of the pharmaceutical compositions of the invention for rectal or vaginal administration may be presented as suppositories, which may be prepared by mixing one or more compounds of the invention with one or more suitable non-irritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which are solid at room temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

Dosage forms of the invention suitable for vaginal administration also include vaginal rings, tampons, creams, gels, pastes, foams or sprays containing such carriers as are known in the art to be suitable.

In the present invention, the pharmaceutical composition or medicament may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Oral administration or injection administration is preferred. The pharmaceutical compositions of the present invention may contain any of the usual non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles. In some cases, pharmaceutically acceptable acids, bases or buffers may be used to adjust the pH of the formulation to improve the stability of the formulated compound or its dosage form in which it is administered. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intracolic, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may be administered to a subject by any route as long as the target tissue is reached.

The term "crystalline" or "crystalline form" refers to a solid form that exhibits substantially three-dimensional ordering. In some embodiments, the crystalline form of the solid is a solid form that is not substantially amorphous. In some embodiments, the crystalline form has an X-ray powder diffraction (XRPD) pattern comprising one or more sharp defined peaks. Different crystalline forms typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystal shape, optoelectronic properties, stability and solubility. Recrystallization solvent, crystallization rate, storage temperature, and other factors may cause a crystalline form to dominate. Various polymorphs of a compound may be prepared by crystallization under different conditions.

The term "co-crystal" refers to a crystal structure comprising at least two different components (e.g., progesterone and a co-form), wherein each component is independently an atom, ion, or molecule. In some embodiments, none of the components are solvents. In some embodiments, at least one component is a solvent. The co-crystal of progesterone and the co-form is different from the salt formed by progesterone and the co-form. In salts, progesterone is complexed with the coformer in such a way that protons are easily transferred from the coformer (e.g., intact proton transfer) to the progesterone at room temperature. However, in this co-crystal, progesterone is complexed with the co-former in such a way that protons are not readily transferred from the co-former to the progesterone at room temperature. In some embodiments, in the co-crystal, no protons are transferred from the conformality to the progestin. In some embodiments, in the co-crystal, a portion of the protons are transferred from the conformality to the progestin. Co-crystals can be used to improve the properties of progesterone (e.g., solubility, stability, ease of formulation, or bioavailability).

The terms "treat," "treating," and "treatment" refer to reversing, alleviating, delaying the onset of, or inhibiting the progression of the diseases described herein. In some embodiments, treatment may be administered after the disease has appeared or one or more signs or symptoms have been observed. In other embodiments, the treatment may be administered without signs or symptoms of the disease. For example, treatment can be administered to a susceptible subject prior to the onset of symptoms (e.g., based on a history of symptoms and/or based on exposure to a pathogen) to delay or prevent the onset of disease. Treatment may also be continued after the symptoms have disappeared, e.g., to delay and/or prevent relapse.

In the prevention and/or treatment of a disease, an effective amount of the co-crystals and pharmaceutical compositions of the present invention are administered to a subject. An "effective amount" of a co-crystal as described herein refers to an amount sufficient to elicit a desired biological response (i.e., to treat and/or reduce the risk of a disorder). As understood by one of ordinary skill in the art, the effective amount of the co-crystals described herein may vary depending on the following factors: biological endpoints, pharmacokinetics of the co-crystal, condition being treated, mode of administration, and age and health of the subject are contemplated. In some embodiments, the effective amount is a therapeutically effective amount. In some embodiments, the effective amount is prophylactic treatment. In some embodiments, an effective amount is the amount of the co-crystal described herein in a single dose. In some embodiments, an effective amount is a combined amount of the co-crystals described herein in multiple doses.

As used herein, a "therapeutically effective amount" is an amount sufficient to provide a therapeutic benefit in treating a disorder or to delay or minimize one or more symptoms associated with the disorder. A therapeutically effective amount of the co-crystal refers to the amount of the therapeutic agent alone or in combination with other therapies to provide a therapeutic benefit in treating the disorder. The term "therapeutically effective amount" can include an amount that improves the overall treatment, reduces or eliminates the symptoms, signs, or causes of the disorder, and/or enhances the efficacy of another therapeutic agent.

A "prophylactically effective amount" as used herein is an amount sufficient to prevent a disorder or one or more symptoms associated with the disorder, or prevent the recurrence thereof. A prophylactically effective amount of a co-crystal refers to an amount of a therapeutic agent that alone, or in combination with other drugs, provides a prophylactic benefit in preventing the condition. The term "prophylactically effective amount" can include an amount that increases the overall prophylactic or prophylactic effect of another prophylactic agent.

Other than in the examples, and where otherwise indicated, all numbers expressing quantities of ingredients used in the specification and claims are to be understood as being modified in all instances by the term "about" or "approximately", such that, unless otherwise indicated, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure, at the very least, and are not intended to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter should be construed in light of the number of significant digits and ordinary rounding approaches.

In the present invention, the diffraction peaks shown in FIG. 5 do not represent an exhaustive list of diffraction peaks exhibited by the crystalline forms. The 2 θ values of the X-ray powder diffraction patterns are slightly variable with machine and with variations in sample preparation and lot-to-lot variations, and the values quoted are not considered absolute values. It should also be understood that the relative intensities of the peaks may vary with orientation effects, and thus the intensities shown in the PXRD traces included in the present disclosure are exemplary and not intended for absolute comparison.

The invention has the advantages and beneficial effects that:

the invention synthesizes the pharmaceutical cocrystal of the progesterone and the pyridine derivative for the first time, improves the stability and the solubility of the progesterone, and improves the drug effect and the bioavailability.

Drawings

FIG. 1 is a TGA/DSC scan of a progesterone-pyridine derivative co-crystal; wherein, fig. 1A and 1B are TGA diagram and DSC diagram of progesterone-2, 6-diaminopyridine eutectic, respectively; FIGS. 1C and 1D are a TGA chart and a DSC chart, respectively, of a progesterone-4-pyridinecarboxamide co-crystal; FIGS. 1E and 1F are a TGA chart and a DSC chart, respectively, of a progesterone-4-aminopyridine co-crystal;

FIG. 2 is a chart of the infrared absorption spectrum of a progesterone-pyridine derivative co-crystal; wherein, fig. 2A is a progesterone-2, 6-diaminopyridine co-crystal; figure 2B is a progesterone-4-pyridinecarboxamide co-crystal; figure 2C is a progesterone-4-aminopyridine co-crystal;

FIG. 3 is a 1H-NMR nuclear magnetic resonance spectrum of a progesterone-pyridine derivative co-crystal; wherein, fig. 3A is progesterone-2, 6-diaminopyridine co-crystal; figure 3B is a progesterone-4-pyridinecarboxamide co-crystal; figure 3C is a progesterone-4-aminopyridine co-crystal;

fig. 4 is a molecular structural diagram of a progesterone-pyridine derivative co-crystal; wherein, fig. 4A is progesterone-2, 6-diaminopyridine co-crystal; figure 4B is a progesterone-4-pyridinecarboxamide co-crystal; figure 4C is a progesterone-4-aminopyridine co-crystal;

figure 5 is an X-powder diffraction pattern of a progesterone-pyridine derivative co-crystal; wherein, fig. 5A and 5B are progesterone-2, 6-diaminopyridine co-crystals; FIGS. 5C and 5D are progesterone-4-pyridinecarboxamide co-crystals; FIGS. 5E and 5F are progesterone-4-aminopyridine co-crystals;

fig. 6 is a scanning electron micrograph of progesterone-pyridine derivative co-crystals; wherein, fig. 6A is progesterone-2, 6-diaminopyridine co-crystal; figure 6B is a progesterone-4-pyridinecarboxamide co-crystal; figure 6C is a progesterone-4-aminopyridine co-crystal;

fig. 7 is a graph of stability experiments for progesterone-pyridine derivative co-crystals; wherein, fig. 7A is progesterone-2, 6-diaminopyridine co-crystal; figure 7B is a progesterone-4-pyridinecarboxamide co-crystal; figure 7C is a progesterone-4-aminopyridine co-crystal;

fig. 8 is a solubility experimental plot of a progesterone-pyridine derivative co-crystal; wherein, fig. 8A is progesterone-2, 6-diaminopyridine co-crystal; figure 8B is a progesterone-4-pyridinecarboxamide co-crystal; figure 8C is a progesterone-4-aminopyridine co-crystal;

fig. 9 is a bioavailability graph of a progesterone-pyridine derivative co-crystal; wherein, fig. 9A is progesterone-2, 6-diaminopyridine co-crystal; figure 9B is a progesterone-4-pyridinecarboxamide co-crystal; figure 9C is a progesterone-4-aminopyridine co-crystal.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. The experimental procedures, in which specific conditions are not specified in the examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.

EXAMPLE 1 preparation of drug Co-crystals

1. Material

Progesterone: zhejiang Xianju pharmaceutical Co Ltd (purity > 99%)

2, 6-diaminopyridine, 4-pyridinecarboxamide, 4-aminopyridine: shanghai Mielin Biochemical technology Co., Ltd. (purity > 99%)

Ethyl acetate: beijing chemical plant (analytical grade) ethyl acetate: beijing chemical plant (analytical pure)

2. Experimental methods

1) Preparation of samples

943.4mg (3.0mmol) of progesterone, 327.4mg (3.0mmol) of 2, 6-diaminopyridine, 366.4mg (3.0mmol) of 4-pyridinecarboxamide, and 282.3mg (3.0mmol) of 4-aminopyridine were weighed and added into a ball mill (planetary ball mill, Nanjing Chin scientific and technological development Co., Ltd.) and ball-milled at 28Hz for 40min without solvent, thus obtaining a white powder sample.

2) Cultivation of single crystals

Taking 100mg of the powder sample, adding 15ml of ethyl acetate to dissolve the powder sample, slowly volatilizing the solvent, and obtaining colorless massive crystals after about 1 week.

EXAMPLE 2 TG-DSC analysis of drug cocrystals

1. Conditions of the experiment

The instrument model is as follows: TGA/DSC 3+

The heating rate is as follows: 10 ℃/min

Temperature range: 40-400 deg.C

Gas atmosphere: nitrogen gas

2. Results

The results are shown in fig. 1, and a Differential Scanning Calorimeter (DSC) shows that the melting point of the eutectic of progesterone-2, 6-diaminopyridine is 111.1 ℃ in the test temperature range; the melting point of the progesterone-4-pyridine carboxamide eutectic is 140.4 ℃; the melting point of the progesterone-4-aminopyridine eutectic is 127.4 ℃;

thermogravimetric analysis (TGA) shows: decomposition of the pyridine derivative and decomposition of progesterone were observed for all co-crystals over the temperature range tested.

EXAMPLE 3 Infrared absorption Spectroscopy (IR) of drug cocrystals

1. Experimental Material

The instrument model is as follows: bruker EQUINOX 55FT-IR

2. Experimental methods

Taking a proper amount of sample, placing the sample in the middle of a probe, and testing

3. Results

As a result, as shown in FIG. 2 and Table 1, the drug cocrystals exhibited infrared absorption peaks at the positions shown in Table 1.

TABLE 1 major infrared absorption peaks of drug cocrystals

EXAMPLE 4 Co-crystals of drugs¹H-NMR nuclear magnetic resonance spectrum

1. Experimental Material

The instrument comprises the following steps: AVANCE III HD 500MHz type nuclear magnetic resonance spectrometer.

Solvent: d6-DMSO (TMS internal standard)

2. Results

The results are shown in FIG. 3, where the shifts of the eutectic NMR spectra are as follows:

progesterone-2, 6-diaminopyridine co-crystal:¹H-NMR(400MHz,d⁶-DMSO)δ＝7.01(t,1H),5.63-5.59(t,3H),5.28(s,4H),2.57-2.55(t,1H),2.42-2.33(m,2H),2.26-2.22(m,2H),2.18-2.14(m,6H),1.80-1.64(d,1H),1.60-1.52(m,5H),1.43-1.35(m,2H),1.20-1.14(m,5H),0.99-0.91(m,2H),0.67(S,3H)。

progesterone-4-pyridinecarboxamide co-crystal:¹H-NMR(400MHz,d⁶-DMSO)δ＝8.72(d,2H),8.24(s,1H),7.78(d,2H),7.71(s,1H),5.63(s,1H),2.57-2.55(t,1H),2.42-2.33(m,2H),2.26-2.22(m,2H),2.18-2.14(m,6H),1.80-1.64(d,1H),1.60-1.52(m,5H),1.43-1.35(m,2H),1.20-1.14(m,5H),0.99-0.91(m,2H),0.67(S,3H)。

progesterone-4-aminopyridine co-crystal:¹H-NMR(400MHz,d⁶-DMSO)δ＝7.97(d,2H),6.45(d,2H),5.96(s,2H),5.64(s,1H),5.63(s,1H),2.57-2.55(t,1H),2.42-2.33(m,2H),2.26-2.22(m,2H),2.18-2.14(m,6H),1.80-1.64(d,1H),1.60-1.52(m,5H),1.43-1.35(m,2H),1.20-1.14(m,5H),0.99-0.91(m,2H),0.67(S,3H)。

EXAMPLE 5X-ray Single Crystal diffraction (SXRD) of drug cocrystals

1. Experimental Material

The instrument model is as follows: rigaku AFC-10

2. Method of producing a composite material

Selecting colorless crystals with size of 0.31mm × 0.22mm × 0.18mm, adopting graphite monochromatized Mo-K ray, and radiating with wavelength

Measuring temperature: 173.00(15) K. SHELL-1 is adopted for structural analysis and refinement4 and Olex2 procedures are completed. The atomic position is determined by a direct method, then all non-hydrogen atomic coordinates are obtained by a difference function method and a least square method, and the structure is corrected by the least square method.

3. Results

As a result, the molar ratio of progesterone to pyridine derivative of the progesterone-pyridine derivative co-crystal was 1:1, as shown in fig. 4 and table 2.

TABLE 2 Crystal Structure

EXAMPLE 6 powder X-ray diffraction Spectroscopy (PXRD) of drug cocrystals

1. Experimental Material

The instrument model is as follows: x-ray powder apparatus model D8 ADVANCE of Bruker, germany.

2. Conditions of the experiment

Copper target, 40KV/40mA, initial angle 5 degree, end angle 60 degree, step width 0.02, scanning speed 17.7 second/step wavelength

A graphite monochromator.

3. Results

The experimental results are shown in fig. 5 and table 3, and the X-ray powder diffraction pattern of the progesterone-pyridine derivative eutectic has characteristic diffraction peaks corresponding to the corresponding positions of the 2 theta values.

TABLE 3 absorption peaks of X-ray powder diffraction spectra of drug cocrystal samples

EXAMPLE 7 scanning Electron microscopy of drug cocrystals

Drug cocrystal samples were scanned using a Jeol JSM-6100 instrument.

The test results are shown in fig. 6, fig. 6A, 6B and 6C are scanning electron microscope images of the progesterone-2, 6-diaminopyridine eutectic, the progesterone-4-pyridinecarboxamide eutectic and the progesterone-4-aminopyridine eutectic respectively, the left image is 500 x, and the right image is 5000 x; as can be seen from a scanning electron microscope image, the size of the eutectic crystal particles is small, and the water solubility is favorably improved; the eutectic particles are uniform, which shows that the preparation process is good and is very beneficial to industrial production and preparation.

EXAMPLE 8 stability Studies of drug Co-Crystal progesterone-pyridine derivatives

1. Experimental Material

Comprehensive medicine stability test box (Shanghai-Hengscientific instruments Co., Ltd.)

2. Experimental methods

The stability experiment examines the stability and the transformation rule of the sample under the conditions of high temperature, high humidity and illumination. 100mg of test sample powder is filled into a weighing bottle, the opening of the sample is respectively placed under the conditions of high temperature (60 +/-2 ℃), high humidity (90% +/-5%) and illumination (4500 +/-500 lx) for 10 days, and samples are respectively taken on the 0 th day, the 5 th day and the 10 th day for powder X-ray diffraction analysis.

2. Results

The experimental results are shown in fig. 7, and the powder X-ray diffraction of the progesterone-2, 6-diaminopyridine eutectic, the progesterone-4-pyridinecarboxamide eutectic and the progesterone-4-aminopyridine eutectic under the conditions of high temperature, high humidity and light does not change significantly, which indicates that the eutectic is stable.

Example 9 solubility study of pharmaceutical Co-Crystal progesterone-pyridine derivatives

1. Experimental Material

Tiandatianfa RC806D dissolution tester

2. Experimental methods

Sieving the sample with a 100-mesh sieve, weighing 2g of the sample, respectively adding into 1000mL of deionized water, stirring at 37 ℃ and 100rpm by a slurry method, respectively sampling 2mL at 5min, 15min, 30min, 45min, 60min, 90min, 120min, 180min, 240min, 300min and 360min, filtering with a filter membrane, taking the filtrate as the sample, injecting 2 needles, and detecting by HPLC.

Liquid phase conditions: high performance liquid chromatograph: waters; chromatographic column of Waters Xbridge C184.6 × 150mm, 3.5 μm chromatographic column; mobile phase: acetonitrile and water 45: 55; the detection wavelength is 280 nm; flow rate: 0.8 mL/min; column temperature: 40 ℃; sample introduction amount: 20 mu L of the solution; operating time: and 15 min.

3. Results of the experiment

The experimental result is shown in fig. 8, and the water solubility of the pharmaceutical co-crystal progesterone-2, 6-diaminopyridine co-crystal, progesterone-4-pyridinecarboxamide co-crystal and progesterone-4-aminopyridine co-crystal is significantly increased.

EXAMPLE 10 pharmacokinetic Studies of drug Co-Crystal progesterone-pyridine derivatives

1. Animal(s) production

Healthy female SD rats (7-9 weeks old, weight 220 + -30 g, 5 per group) were purchased from Peking Wintolite laboratory animals technology, Inc.

2. Animal administration method

All animals were kept on a 12 hr/12 hr light/dark cycle, 5 animals per cage, with free diet. The progesterone eutectic is prepared into a solution with PBS (phosphate buffered saline, containing 0.1% DMSO), the dosage of the progesterone eutectic is 5.0mg/kg, and the progesterone eutectic is administered through intramuscular injection.

3. Method for preparing biological sample

Samples of 50 μ L jugular venous blood were collected in batches at defined times (15, 30min and 1, 1.5, 2, 3, 5, 7, 12, 24, 48h) into heparinized tubes. The blood was then centrifuged for 15 min. Plasma samples were stored at-20 ℃. mu.L rat plasma, 5. mu.L methanol and 200. mu.L internal standard solution (buspirone, 5ng/mL) were added to 1mL methanol: acetonitrile (1:1, v/v). The plasma samples were vortexed for 1min, centrifuged at 4000rpm for 15min, and the plasma samples were vortexed with methanol: the supernatant was diluted 20 times with water (1:1, v/v, 0.1% trifluoroacetic acid) and injected.

4. Biological sample analysis method

LC-MS/MS instrument model: AB Sciex 5500; LC-MS/MS quantitative analysis software: 1.6.3; an ionization mode: electrospray positive ions; the scanning mode is as follows: multiple Reaction Monitoring (MRM); analyte MRM: progesterone-DNS, 530.4/171.0; internal standard MRM: buspirone, 386.2/122.2;

liquid phase conditions: shimadzu LC-30AD, ACE Excel 5C4(50mm × 2.1mm), sample size 10 μ L; mobile phase: a5 mM ammonium acetate (0.05% trifluoroacetic acid) and B acetonitrile (0.1% trifluoroacetic acid) at a flow rate of 0.8mL/min, and the gradients of mobile phases A and B are shown in the following table.

TABLE 4 gradient of mobile phases A and B

Time (min)	A(％)	B(％)
			0.4-1.8	80	20
1.8-2.7	5	95
			2.7-3.5	80	20

5. Results

The test results are shown in FIG. 9, where the peak reaching time of progesterone is 1h, the peak reaching time of co-crystal is 1.5h, and the co-crystal C of progesterone-2, 6-diaminopyridine_max1.6 times of progesterone; progesterone-4-pyridinecarboxamide cocrystalsC_max1.6 times of progesterone; progesterone-4-aminopyridine cocrystal C_maxIs 1.7 times of progesterone, and has better bioavailability.

The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.

Claims

1. Co-crystal of progesterone and a co-form, characterized in that said co-form is selected from 2, 6-diaminopyridine, the empirical formula of the co-crystal of progesterone-2, 6-diaminopyridine being C₂₆H₃₇N₃O₂The co-crystal has an X-ray spectrum with characteristic 2 θ values at 2 θ ± 0.2 ° as follows: 13.402, 16.92, 19.279, 19.957, 25.241.

2. The co-crystal of claim 1, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 10.459, 14.799, 15.078, 18.679, 23.458, 23.879.

3. The co-crystal of claim 2, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 8.539, 15.693, 20.919, 21.139, 23.138, 26.96, 29.737, 31.602.

4. The co-crystal of any one of claims 1-3, wherein the X-ray powder diffraction pattern is substantially as shown in FIG. 5B.

5. The co-crystal of claim 1, wherein the co-crystal comprises an endothermic peak at a melting point temperature of about 111.1 ℃.

6. The co-crystal of claim 5, wherein the thermogram is substantially as shown in fig. 1B.

7. The co-crystal of claim 1, wherein the co-crystal comprises the following crystallographic parameters:

the progesterone-2, 6-diaminopyridine eutectic is monoclinic system, P2₁Space group, unit cell parameters a = 13.0819(5) a, b = 18.6192(6) a, c = 19.7511(8) (11) a, α = γ =90.00^o，β= 107.375(4)^oZ =8, unit cell volume 4591.3(3) A³。

8. Co-crystals of progesterone and a co-form, characterized in that said co-form is selected from the group consisting of 4-pyridinecarboxamide, the empirical formula of the co-crystals of progesterone-4-pyridinecarboxamide being C₂₇H₃₆N₂O₃The co-crystal has an X-ray spectrum with characteristic 2 θ values at 2 θ ± 0.2 ° as follows: 13.5, 20.421.

9. The co-crystal of claim 8, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 13.1, 15.022, 17.401, 18.282, 22.218.

10. The co-crystal of claim 9, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 21.92, 26.418, 27.516.

11. The co-crystal of any one of claims 8-10, wherein the X-ray powder diffraction pattern is substantially as shown in fig. 5D.

12. The co-crystal of claim 8, wherein the co-crystal comprises an endotherm at a melting point temperature of about 140.4 ℃.

13. The co-crystal of claim 12, wherein the thermogram is substantially as shown in fig. 1D.

14. The co-crystal of claim 8, wherein the co-crystal comprises the following crystallographic parameters:

the progesterone-4-pyridine carboxamide eutectic is orthorhombic, P2₁2₁2₁Space group, unit cell parameters a = 7.8943(5) a, b = 12.9909(17) a, c = 46.685(2) a, α = β = γ =90.00^oZ =8, unit cell volume 4787.7(7) A³。

15. Co-crystals of progesterone and a co-form, characterized in that said co-form is selected from 4-aminopyridine, the empirical formula of the co-crystals of progesterone-4-aminopyridine being C₂₆H₃₆N₂O₂The co-crystal has an X-ray spectrum with characteristic 2 θ values at 2 θ ± 0.2 ° as follows: 13.717, 15.899, 16.079, 21.441.

16. The co-crystal of claim 15, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 14.3, 15.499, 18.499, 19.321, 19.92, 26.041, 29.64.

17. The co-crystal of claim 16, further having an X-ray spectrum with characteristic 2 Θ values at 2 Θ ± 0.2 ° as follows: 17.382, 22.619, 25.599, 28.721.

18. The co-crystal of any one of claims 15-17, wherein the X-ray powder diffraction pattern is substantially as shown in fig. 5F.

19. The co-crystal of claim 15, wherein the co-crystal comprises an endotherm at a melting point temperature of about 127.4 ℃.

20. The co-crystal of claim 19, wherein the thermogram is substantially as shown in fig. 1F.

21. The co-crystal of claim 15, wherein the co-crystal comprises the following crystallographic parameters:

the progesterone-4-aminopyridine eutectic is orthorhombic, P2₁2₁2₁Space group, unit cell parameters a = 7.6234(6) a, b = 12.2741(8) a, c = 24.4042(16) a, α = β = γ =90.00 a^oZ =4, unit cell volume 2283.5(3) A³。

22. A method of preparing a co-crystal according to any one of claims 1 to 21, comprising:

23. The method of claim 22, wherein the solvent is ethyl acetate.

24. The method of claim 22, wherein the molecular ratio of progesterone to co-form is 1: 1.

25. The method of claim 22, wherein the progesterone and the co-form are pulverized using a ball mill.

26. A pharmaceutical composition comprising a co-crystal according to any one of claims 1 to 21.

27. The pharmaceutical composition of claim 26, further comprising a pharmaceutically acceptable carrier.

28. The pharmaceutical composition of claim 26 or 27, wherein the pharmaceutical composition is administered orally, by injection, vaginally, or transdermally.

29. Use of a co-crystal according to any one of claims 1 to 21 or a pharmaceutical composition according to any one of claims 26 to 28 in the manufacture of a medicament for maintaining pregnancy.

30. Use of a co-crystal according to any one of claims 1 to 21 or a pharmaceutical composition according to any one of claims 26 to 28 in the manufacture of a medicament for the prevention and/or treatment of gynaecological disorders.

31. Use according to claim 30, wherein the gynaecological disorder comprises spontaneous preterm birth, luteal insufficiency, secondary amenorrhea, premenstrual syndrome.

32. Use of the co-crystal of any one of claims 1 to 21 or the pharmaceutical composition of any one of claims 26 to 28 for the preparation of a medicament for the prevention and/or treatment of cardiovascular diseases, neurological diseases.