Neurosteroid derivatives and uses thereof

Abstract

The present invention provides a neurosteroid derivative having a structure shown in formula (I) and its use. Compared with the prior art, the neurosteroid derivative having a structure shown in formula (I) provided by the present invention has significantly higher bioavailability and can be administered via non-intravenous administration for the treatment and / or prevention of GABA-A receptor-mediated diseases. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of medicinal chemistry, in particular to a class of neurosteroid derivatives and their medical uses. Background Art

[0002] GABA (γ-aminobutyric acid) is the main neurotransmitter in the central nervous system. It can increase the flow of ions inside and outside the nerve cell membrane by mediating the downstream ionotropic channel GABA-A receptor and exert physiological effects. The active site of GABA-A receptor can bind to GABA and many drugs such as muscimol, gaboxadol, bicuculline, etc. There are many isomeric modulators that can indirectly regulate the activity of GABA-A receptors, including benzodiazepines Class, barbiturates, inhalation anesthetics, etc.

[0003] Neurosteroids are steroid hormone derivatives that do not have hormonal effects, but they still affect neuronal excitability by regulating ionotropic receptors and can bind to GABA-A receptors. Dehydroepiandrosterone, progesterone, and their metabolites can regulate the expression of GABA-A receptor subunits through genetic mechanisms, thereby affecting neurotransmitters. Neurosteroids have fewer restrictions on subunits and are more effective than benzodiazepines. The application range of this class is wider. The neurosteroid drugs currently developed and / or under development targeting GABA-A receptors include allopregnanolone, ganaxolone, antacidone, alfadolon, etc.

[0004] Allopregnanolone, also known as 5α-pregnane-3α-ol-20-one, is an endogenous inhibitory pregnane neurosteroid. The effects of allopregnanolone are similar to those of other positive allosteric modulators of GABA action at GABA-A receptors, such as the benzodiazepines Endogenously produced allopregnanolone plays a key neurophysiological role by fine-tuning GABA-A receptors and modulating the effects of several positive allosteric modulators and agonists on GABA-A receptors.

[0005] Ganaxolone, also known as 3α-hydroxy-3-methyl-5α-pregnane-20-one, is a synthetic neurosteroid analog that acts as a positive allosteric modulator of GABA-A receptors. Ganaxolone shows promise in treating temporal lobe epileptic seizures as well as catamenial epilepsy. Ganaxolone is also being studied for the treatment of post-traumatic stress disorder, fragile X-syndrome, neuralgic headaches, neonatal seizures, and postpartum depression. Ganaxolone has been approved by the U.S. FDA for the treatment of epilepsy in women with protocadherin-19 gene (PCDH19) and is well tolerated in adults and children.

[0006] GABA-A receptor positive allosteric modulators including allopregnanolone and ganaxolone have defects such as low water solubility and poor metabolic stability, which lead to insufficient bioavailability of the drug in the human body, and bring limitations to its clinical application. At present, the structural modification of GABA-A receptor positive allosteric modulators including allopregnanolone and ganaxolone is mostly focused on improving water solubility, thereby promoting drug absorption, improving bioavailability and pharmacokinetic properties. For example, CN108148106A discloses a class of water-soluble pregnenolone derivatives and their uses, wherein the water-soluble pregnenolone derivatives have good physical / chemical stability and good water solubility, and can be quickly decomposed and released in plasma. Active drugs (allopregnanolone), and then quickly produce pharmacological effects. There are also cases where oral administration is achieved by structurally modifying GABA-A receptor positive allosteric modulators to reduce their metabolic rate. For example, CN109503694A discloses a novel GABA-A receptor modulator, which reduces the metabolic rate of the compound by introducing an aromatic group or a heteroaromatic group next to the 20-carbonyl group of allopregnanolone, maintains good biological activity, and achieves oral administration.

[0007] At present, there is an urgent clinical need for GABA-A receptor modulators with good bioavailability, especially metabolic stability in vivo. The purpose of the present invention is to find drugs with better bioavailability in vivo, especially better metabolic stability in vivo, by structurally modifying existing neurosteroid drugs. Summary of the invention

[0008] In one aspect, the present invention provides a compound represented by formula (I):

[0009]

[0010] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof, wherein R1 and R2 are as defined herein.

[0011] In one aspect, the present invention provides a method for preparing a compound of formula (I) or its stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts.

[0012] On the other hand, the present invention provides a pharmaceutical composition for treating and / or preventing central nervous system diseases, comprising a compound having a structure shown in formula (I) or its stereoisomers, tautomers, isotopically enriched analogs, solvates, pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0013] On the other hand, the present invention also provides the use of a compound having a structure represented by formula (I) or its stereoisomers, tautomers, isotope-enriched analogs, solvates and pharmaceutically acceptable salts in the preparation of drugs for treating and / or preventing central nervous system diseases.

[0014] On the other hand, the present invention also provides a method for treating and / or preventing central nervous system diseases, comprising administering to an individual in need thereof a therapeutically effective amount of a compound having a structure represented by formula (I) or its stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts.

[0015] Beneficial effects: Compared with the prior art, the compounds of the present invention or their stereoisomers, tautomers, isotope-enriched analogs, solvates and pharmaceutically acceptable salts thereof exhibit significantly better pharmacokinetic properties, especially in vivo metabolic stability, and can be administered via non-intravenous administration for the treatment of a variety of central nervous system diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention.

[0017] Figure 1 The results show that the compound 1 of the present invention has a regulatory effect on the GABA-A (α4β3δ) receptor. DETAILED DESCRIPTION

[0018] definition

[0019] As used in this specification, the following words and phrases are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0020] As used herein, the term "alkyl" refers to a monovalent group of a branched or unbranched saturated hydrocarbon chain having 1 to 12 carbon atoms, more typically 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 6 carbon atoms. This term is exemplified by groups such as methyl, ethyl, 1-propyl (n-propyl), 2-propyl (isopropyl), 1-butyl (n-butyl), 2-methyl-1-propyl (isobutyl), 2-butyl (sec-butyl), 2-methyl-2-propyl (tert-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, 1-heptyl, 1-octyl, 1-nonyl, 1-decyl, and the like.

[0021] As used herein, the term "alkenyl" refers to a monovalent straight or branched unsaturated hydrocarbon group having the indicated number of carbon atoms (e.g., 2 to 12 carbon atoms, 2 to 10 carbon atoms, 2 to 8 carbon atoms, or 2 to 6 carbon atoms, etc.) and having carbon-carbon double bonds (e.g., 1, 2 or 3 carbon-carbon double bonds). In some embodiments, examples of alkenyl include, but are not limited to, vinyl (i.e., -CH=CH2), propen-1-yl (i.e., -CH=CHCH3), propen-3-yl (or allyl, i.e., -CH2CH=CH2), propen-2-yl (i.e., -C(CH3)=CH2), butadienyl (including 1,2-butadienyl and 1,3-butadienyl), and the like.

[0022] As used herein, the term "alkynyl" refers to a monovalent straight or branched unsaturated hydrocarbon group having the indicated number of carbon atoms (e.g., 2 to 12 carbon atoms, 2 to 10 carbon atoms, 2 to 8 carbon atoms, or 2 to 6 carbon atoms, etc.) and having a carbon-carbon triple bond (e.g., 1, 2 or 3 carbon-carbon triple bonds). In some embodiments, examples of alkynyl include, but are not limited to, ethynyl (i.e., -C≡CH), propargyl (i.e., -CH2C≡CH), propynyl (i.e., -C≡CCH3), etc.

[0023] As used herein, the term "cycloalkyl" refers to a monovalent saturated carbocyclic group of 3 to 12 carbon atoms (more typically 3 to 10 carbon atoms, 3 to 8 carbon atoms, or 3 to 6 carbon atoms) having a single ring or multiple fused or bridged rings. In some embodiments, the cycloalkyl group includes a monocyclic structure, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, etc., or a polycyclic structure, such as adamantyl and bicyclo[2.2.1]heptanyl, or a cycloalkyl group fused to an aryl group, such as indan, etc., if the point of attachment is through the cycloalkyl group.

[0024] As used herein, the term "aryl" refers to an aromatic carbocyclic group of 6 to 14 carbon atoms (more typically 6 to 10 carbon atoms, or 6 carbon atoms) having a single ring (e.g., phenyl) or multiple rings (e.g., biphenyl) or multiple condensed (fused) rings (e.g., naphthyl, fluorenyl, and anthracenyl). The term is exemplified by groups such as phenyl, fluorenyl, naphthyl, anthracenyl, 1,2,3,4-tetrahydronaphthalene (if the point of attachment is through the aryl group), and the like.

[0025] As used herein, the term "heteroaryl" refers to an aromatic ring group containing a monocyclic or multiple condensed (fused) ring (e.g., containing 2 or 3 rings) of 5 to 14 ring atoms in the ring, wherein in addition to carbon atoms, the ring atoms also include at least one heteroatom selected from oxygen, nitrogen and / or sulfur. If the ring is aromatic, sulfur and nitrogen atoms can also exist in oxidized form. Multiple condensed (fused) ring heteroaryl is a monocyclic heteroaryl as defined above fused with one or more rings selected from the following to form a multiple condensed ring system: heteroaryl (to form, for example, naphthyridinyl, such as 1,8-naphthyridinyl), heterocycle (for example, 1,2,3,4-tetrahydronaphthyridinyl, such as 1,2,3,4-tetrahydro-1,8-naphthyridinyl), carbocycle (to form, for example, 5,6,7,8-tetrahydroquinolinyl) and aryl (to form, for example, indazolyl). Such multiple fused ring systems can be optionally substituted with one or more (e.g., 1, 2, 3, or 4) oxo groups on the carbocyclic or heterocyclic portions of the fused rings. When valence requirements permit, the rings of the multiple fused ring systems can be interconnected by fusion, spirocyclic, and bridging bonds. It should be understood that the individual rings of the multiple fused ring systems can be connected to each other in any order. It should also be understood that the attachment point of the multiple fused ring system can be at any position of the multiple fused ring system, including heteroaryl, heterocyclic, aryl, or carbocyclic portions of the multiple fused system. It should also be understood that the attachment point of the heteroaryl group can be on any suitable atom of the heteroaryl group, including carbon atoms and heteroatoms (e.g., nitrogen). Exemplary heteroaryl groups include, but are not limited to, pyridinyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazolyl, thienyl, indolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, furanyl, oxadiazolyl, thiadiazolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, indazolyl, quinoxalinyl, quinazolinyl, 5,6,7,8-tetrahydroisoquinolinyl, benzofuranyl, benzimidazolyl, thiaindenyl, pyrrolo[2,3-b]pyridinyl, quinazolinyl-4(3H)-one, triazolyl, 4,5,6,7-tetrahydro-1H-indazolyl, and 3b,4,4a,5-tetrahydro-1H-cyclopropane[3,4]cyclopenta[1,2-c]pyrazolyl.

[0026] As used herein, the term "heterocyclyl" refers to a monocyclic saturated or partially unsaturated group having a 3-8-membered monocyclic ring or multiple condensed (fused) rings or bridged rings with 3 to 14 ring atoms in the ring, wherein in addition to carbon atoms, the ring atoms also contain at least one heteroatom selected from oxygen, nitrogen and / or sulfur. Examples of heterocyclyl groups include, but are not limited to, tetrahydrofuranyl, morpholinyl, piperidinyl, piperazinyl, dihydropyridinyl, 4,5,6,7-tetrahydro-1H-benzo[d]imidazolyl, benzo[d]imidazolyl, 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridinyl, aziridine, azetidine, tetrahydropyrrolyl, azepanyl, azooctanyl, Oxirane, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, oxepanyl, oxocanyl, thiirane, thietanyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, thiepanyl, thiocanyl, tetrahydroimidazolyl, tetrahydropyrazolyl, tetrahydrooxazolyl, tetrahydroisoxazolyl, tetrahydrothiazolyl, tetrahydroisothiazolyl, dioxanyl, thioxanyl, dithianyl and the like.

[0027] As used herein, the term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0028] As used herein, the term "alkoxy" refers to an "alkyl-O-" group, wherein alkyl is as defined herein. This term is exemplified by groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, and the like.

[0029] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to affect treatment as defined below when administered to a mammal in need of such treatment. The therapeutically effective amount will vary with the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the mode of administration, etc., and can be readily determined by one of ordinary skill in the art.

[0030] As used herein, the term "stereoisomer" refers to a compound having the same chemical composition and connectivity, but whose atoms have different orientations in space that cannot be interchanged by single bond rotation. "Stereoisomer" includes "diastereomers" and "enantiomers". "Diastereomers" refers to stereoisomers with two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral characteristics and reactivity. Diastereomeric mixtures can be separated under high-resolution analytical procedures (such as crystallization, electrophoresis and chromatography). "Enantiomers" refers to two stereoisomers that are non-overlapping mirror images of each other.

[0031] As used herein, the term "tautomer" refers to the coexistence of two (or more) compounds that differ only in the position and electron distribution of one (or more) mobile atoms, such as keto-enol tautomers.

[0032] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0033] Any formula or structure given herein, including Formula I or any formula disclosed herein, is also intended to represent unlabeled forms of the compound as well as isotopically labeled forms. These forms of the compound may also be referred to as "isotopically labeled" or "isotopically enriched analogs." An isotopically labeled compound has a structure as depicted herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, such as, but not limited to, 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogens have been replaced with deuterium.

[0034] As used herein, the term "solvate" refers to an association or complex of one or more solvent molecules with a compound of the invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine. The term "hydrate" refers to a complex in which the solvent molecule is water.

[0035] Compound

[0036] In one embodiment, the present invention provides a compound having a structure shown in the following formula (I):

[0037]

[0038] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof, wherein:

[0039] R1 is selected from hydrogen or C1-C 12 alkyl;

[0040] R2 is selected from R-(C=O)-, R-(C=S)-, R-(S=O)-, R-(SO2)-, R-CH(OH)-, RO-, RS-, NO2, wherein R is selected from hydrogen, C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 12 Cycloalkyl, C6-C 14 Aryl, heteroaryl, heterocyclic, NR3R3', wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclic is optionally substituted by halogen, cyano, nitro, hydroxyl, C1-C 12 Alkoxy, C1-C 12 Alkoxycarbonyl, C1-C 12 Alkoxycarbonyloxy, C1-C 12 Alkyl acyloxy, carboxyl, thiol, C1-C 12 alkylthio, NR4R4'substituted; R3, R3', R4, R4' are each independently selected from hydrogen, C1-C 12 Alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C3-C 12 Cycloalkyl, C6-C 14 An aryl group, a heteroaryl group, a heterocyclic group, or R3 and R3', R4 and R4' together with the nitrogen atom to which they are attached form a heterocyclic group.

[0041] In one embodiment, in the compound having the structure shown in the following formula (I) provided by the present invention, or its stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts, R1 is selected from hydrogen or C1-C 12 Alkyl; preferably, R1 is selected from hydrogen or C1-C6 alkyl; more preferably, R1 is selected from hydrogen or methyl; most preferably, R1 is selected from hydrogen.

[0042] In one embodiment, in the compound having the structure shown in the following formula (I) provided by the present invention, or its stereoisomers, tautomers, isotope-enriched analogs, solvates and pharmaceutically acceptable salts, R2 is selected from R-(C=O)-, R-(C=S)-, R-(S=O)-, R-(SO2)-, R-CH(OH)-, RO-, RS-, NO2, wherein R is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C 10 wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl is optionally substituted with halogen, cyano, nitro, hydroxyl, C1-C6 alkoxy, C1-C6 alkoxycarbonyl, C1-C6 alkoxycarbonyloxy, C1-C6 alkylacyloxy, carboxyl, sulfhydryl, C1-C6 alkylthio, NR4R4'; R3, R3', R4, R4' are each independently selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C 10 An aryl group, a heteroaryl group, a heterocyclic group, or R3 and R3', R4 and R4' together with the nitrogen atom to which they are attached form a heterocyclic group.

[0043] In a preferred embodiment, in the compound having the structure shown in the following formula (I) provided by the present invention, or its stereoisomers, tautomers, isotope-enriched analogs, solvates and pharmaceutically acceptable salts, R2 is selected from R-(C=O)-, R-(C=S)-, R-(S=O)-, R-(SO2)-, R-CH(OH)-, RO-, RS-, NO2, wherein R is selected from hydrogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl, phenyl, heteroaryl, heterocyclic group, NR3R3', wherein the alkyl , alkenyl, alkynyl, cycloalkyl, phenyl, heteroaryl, heterocyclyl are optionally substituted by halogen, cyano, nitro, hydroxyl, C1-C4 alkoxy, C1-C4 alkoxycarbonyl, C1-C4 alkoxycarbonyloxy, C1-C4 alkylacyloxy, carboxyl, thiol, C1-C4 alkylthio, NR4R4'; R3, R3', R4, R4' are each independently selected from hydrogen, C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, C3-C6 cycloalkyl, phenyl, heteroaryl, heterocyclyl, or R3 and R3', R4 and R4' together with the nitrogen atom to which they are connected form a heterocyclyl.

[0044] In a preferred embodiment, in the compound having the structure shown in the following formula (I) provided by the present invention, or its stereoisomers, tautomers, isotope-enriched analogs, solvates and pharmaceutically acceptable salts, R1 is selected from hydrogen or C1-C6 alkyl; R2 is selected from R-(C=O)-, R-(C=S)-, R-(S=O)-, R-(SO2)-, R-CH(OH)-, RO-, RS-, NO2, wherein R is selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C 10 wherein the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heterocyclyl is optionally substituted with halogen, cyano, nitro, hydroxyl, C1-C6 alkoxy, C1-C6 alkoxycarbonyl, C1-C6 alkoxycarbonyloxy, C1-C6 alkylacyloxy, carboxyl, sulfhydryl, C1-C6 alkylthio, NR4R4'; R3, R3', R4, R4' are each independently selected from hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, C6-C 10 An aryl group, a heteroaryl group, a heterocyclic group, or R3 and R3', R4 and R4' together with the nitrogen atom to which they are attached form a heterocyclic group.

[0045] In a preferred embodiment, the present invention provides a compound having a structure shown in the following formula (II):

[0046]

[0047] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof, wherein R1 and R2 are as defined herein.

[0048] In a preferred embodiment, the present invention provides a compound having a structure shown in the following formula (III):

[0049]

[0050] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof, wherein R1 and R2 are as defined herein.

[0051] In a preferred embodiment, the present invention provides a compound having a structure shown in the following formula (IV):

[0052]

[0053] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof, wherein R1 and R2 are as defined herein.

[0054] In a preferred embodiment, the present invention provides a compound having a structure shown in the following formula:

[0055]

[0056] or stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts thereof.

[0057] Pharmaceutical compositions and administration

[0058] The compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, prodrug, solvate provided by the present invention is usually administered in the form of a pharmaceutical composition. The present invention therefore provides a pharmaceutical composition, which comprises a compound provided by the present invention as an active ingredient and one or more pharmaceutically acceptable carriers. The pharmaceutical composition can be administered alone or in combination with other therapeutic agents. This composition is prepared in a manner well known in the art (e.g., Reminton's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, PA 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (GS Banker & C.T. Rhodes, Eds.).

[0059] Pharmaceutically acceptable carriers can be solid or liquid. Wherein, solid carriers can be one or more substances used as excipients, diluents, sweeteners, solubilizers, lubricants, adhesives, tablet disintegrating agents, stabilizers, preservatives or encapsulating materials. Liquid carriers can be solvents or liquid dispersion media. Suitable solid carriers include, but are not limited to, for example, cellulose, glucose, lactose, mannitol, magnesium stearate, magnesium carbonate, sodium carbonate, saccharin sodium, sucrose, dextrin, talc, starch, pectin, gelatin, tragacanth gum, gum arabic, sodium alginate, parabens, methylcellulose, sodium carboxymethylcellulose, low melting point wax, cocoa butter, etc. Suitable liquid carriers include, but are not limited to, water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil), glycerides, agar, pyrogen-free water, isotonic saline, Ringer's solution and mixtures thereof.

[0060] The pharmaceutical composition according to the present invention can be in any form suitable for the intended method of administration. For example, when used for oral use, it can be prepared into tablets, lozenges, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs, solutions, sprays. The composition intended for oral use can be prepared according to any method known in the art for making pharmaceutical compositions. The pharmaceutical composition of the present invention can be in the form of a sterile injection preparation, such as a sterile injection aqueous or oily suspension. The pharmaceutical composition of the present invention can also be prepared into a preparation suitable for intrapulmonary or intranasal administration, such as a preparation, nasal drops or nasal sprays for aerosol or dry powder administration. The pharmaceutical composition of the present invention can also be prepared into a suppository suitable for rectal administration. The pharmaceutical composition of the present invention can also be prepared into a transdermal preparation for topical administration or an eye drop suitable for ocular administration.

[0061] The pharmaceutical composition of the present invention can be administered via arterial injection, intravenous injection, intraperitoneal injection, parenteral injection, intramuscular injection, subcutaneous injection, sublingual injection, oral administration, topical administration in single doses or multiple doses.

[0062] The effective dose of the compounds of the present invention depends at least on the nature of the condition being treated, toxicity, delivery method and pharmaceutical formulation, and will be determined by a clinician using conventional dose escalation studies. It can be expected that about 0.0001 to about 100 mg per kilogram of body weight per day; generally about 0.01 to about 10 mg per kilogram of body weight per day; more typically, about 0.01 to about 5 mg per kilogram of body weight per day; most typically about 0.05 to about 0.5 mg per kilogram of body weight per day. For example, the daily candidate dose for an adult of about 70 kg body weight will be in the range of 1 mg to 1000 mg, preferably in the range of 5 mg to 500 mg, and can be in the form of a single dose or multiple doses.

[0063] Indications

[0064] The compounds of the present invention or their stereoisomers, tautomers, isotope-enriched analogs, solvates can be used to prevent and / or treat GABA-A receptor-mediated diseases. In particular, the compounds of the present invention or their stereoisomers, tautomers, isotope-enriched analogs, solvates can be used to prevent and / or treat central nervous system diseases. In particular, the compounds of the present invention or their stereoisomers, tautomers, isotopically enriched analogs, solvates can be used to treat and / or prevent diseases including, but not limited to, postpartum depression, depression, major depressive disorder, bipolar disorder, mood disorders, anxiety, post-traumatic stress disorder (PTSD), premenstrual dysphoric disorder (PMDD), premenstrual syndrome, generalized anxiety disorder, seasonal affective disorder (SAD), social anxiety disorder, memory loss, stress intolerance, Niemann-Pick disease type C or related neurological or physical symptoms, epilepsy, essential tremor, epileptiform disorders, NMDA hypofunction, migraine, status epilepticus, sleep disorders, fragile X syndrome, 5α-reductase inhibitor-induced depression, PCDH19 female pediatric epilepsy, sexual dysfunction, cognitive disorders, Parkinson's disease or Alzheimer's disease.

[0065] Combination medication

[0066] The compounds of the present invention can be administered as a monotherapy to treat and / or prevent GABA-A receptor-mediated diseases, or central nervous system diseases, or diseases selected from postpartum depression, depression, major depressive disorder, bipolar disorder, mood disorders, anxiety, post-traumatic stress disorder (PTSD), premenstrual dysphoric disorder (PMDD), premenstrual syndrome, generalized anxiety disorder, seasonal affective disorder (SAD), social anxiety disorder, memory loss, stress intolerance, Niemann-Pick disease type C or related neurological or physical symptoms, epilepsy, essential tremor, epileptiform disorders, NMDA hypofunction, migraine, status epilepticus, sleep disorders, fragile X syndrome, 5α-reductase inhibitor-induced depression, PCDH19 female pediatric epilepsy, sexual dysfunction, Parkinson's disease or Alzheimer's disease, etc. The compounds of the present invention can also be administered in combination with one or more additional therapeutic agents to treat the diseases mentioned herein. The additional therapeutic agent(s) include, but are not limited to, neuroactive steroids such as pregnanolone, allopregnanolone, alfadolone, ganaxolone, alfaxalone; benzodiazepines Barbiturates.

[0067] Treatment methods and uses

[0068] In one embodiment, the present invention provides a method for treating and / or preventing a GABA-A receptor-mediated disease, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound of the present invention or its stereoisomer, tautomer, isotopically enriched analog, solvate, or a pharmaceutical composition comprising the same. In another embodiment, the present invention provides a method for treating and / or preventing a central nervous system disease, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound of the present invention or its stereoisomer, tautomer, isotopically enriched analog, solvate, and pharmaceutically acceptable salt, or a pharmaceutical composition comprising the same. In another embodiment, the present invention provides a method for treating and / or preventing diseases including postpartum depression, depression, major depressive disorder, bipolar disorder, mood disorders, anxiety, post-traumatic stress disorder (PTSD), premenstrual dysphoric disorder (PMDD), premenstrual syndrome, generalized anxiety disorder, seasonal affective disorder (SAD), social anxiety disorder, memory loss, stress intolerance, Niemann-Pick disease type C or related neurological or physical symptoms, epilepsy, essential tremor, epileptiform disorders, NMDA hypofunction, migraine, status epilepticus, sleep disorders, fragile X syndrome, 5α-reductase inhibitor-induced depression, PCDH19 female pediatric epilepsy, sexual dysfunction, cognitive disorders, Parkinson's disease or Alzheimer's disease, the method comprising administering to an individual in need thereof a therapeutically effective amount of a compound of the present invention or its stereoisomers, tautomers, isotopically enriched analogs, solvates and pharmaceutically acceptable salts, or a pharmaceutical composition comprising the same.

[0069] The present invention provides the use of the compounds of the present invention or their stereoisomers, tautomers, isotopically enriched analogs, and solvates as active therapeutic substances in the preparation of drugs for treating and / or preventing GABA-A receptor-mediated diseases. More particularly, the present invention provides the use of the compounds of the present invention or their stereoisomers, tautomers, isotopically enriched analogs, and solvates as active therapeutic substances in the preparation of drugs for treating and / or preventing central nervous system diseases. More particularly, the present invention provides the use of a compound of the present invention or a stereoisomer, tautomer, isotopically enriched analog, or solvate thereof as an active therapeutic substance in the preparation of a medicament for treating and / or preventing the following diseases, including but not limited to postpartum depression, depression, major depressive disorder, bipolar disorder, mood disorders, anxiety, post-traumatic stress disorder (PTSD), premenstrual dysphoric disorder (PMDD), premenstrual syndrome, generalized anxiety disorder, seasonal affective disorder (SAD), social anxiety disorder, memory loss, stress intolerance, Niemann-Pick disease type C or related neurological or physical symptoms, epilepsy, essential tremor, epileptiform disorders, NMDA hypofunction, migraine, status epilepticus, sleep disorders, fragile X syndrome, 5α-reductase inhibitor-induced depression, PCDH19 female pediatric epilepsy, sexual dysfunction, cognitive disorders, Parkinson's disease or Alzheimer's disease and the like.

[0070] General synthetic method

[0071] Compounds of the present invention can be prepared using methods disclosed herein and their modified approaches and methods well known in the art. Typical embodiments of compounds according to the present invention can be synthesized using the following general reaction schemes. It is apparent from the description herein that corresponding different products can be obtained by replacing reaction raw materials with other materials having similar structures. Reaction raw materials are typically obtained from commercial sources or synthesized using disclosed methods.

[0072] Reaction process

[0073]

[0074] The compound represented by formula (Ia) is reacted with an acid anhydride compound (such as acetic anhydride, trifluoroacetic anhydride, etc.) and nitric acid in a suitable solvent to obtain the compound represented by formula (I).

[0075] The following examples are provided to illustrate the preparation of compounds of the present invention but are not intended to limit the invention in any way.

[0076] Example

[0077] Example 1 Synthesis of Compound 1

[0078]

[0079] Add 5.00 g of allopregnanolone and 105.0 ml of dichloromethane to a 250 ml three-necked flask, stir to dissolve, cool to below -10°C, and drop 45.0 ml of acetic anhydride. Slowly drop 10 ml of nitric acid below -10°C, and continue to react at -10°C until complete. Pour the reaction solution into 300 ml of water, stir, wash the organic phase with 200 ml of water twice, distill the organic phase under reduced pressure, and separate the residue by column chromatography to obtain an off-white solid (4.86 g, yield 85%).

[0080] 1 H NMR (400 MHz, chloroform-d) δ 2.53 (t, J = 9.0 Hz, 1H), 2.39 (td, J = 14.5, 13.6, 6.5 Hz, 1H), 2.34–2.26 (m, 2H), 2.25–2.15 (m, 1H), 2.12 (s, 4H), 2.04 (dtt, J = 12.7, 6.2, 3.2 Hz, 3H), 1.76–1.61 (m, 5H), 1.61–1.49 (m ,1H),1.48–1.40(m,3H),1.40–1.32(m,3H),1.26(ddd,J=23.2,9.9,4.4Hz,2H),1.17(ddd,J=16.4, 9.5, 4.2Hz, 2H), 1.01 (s, 3H), 0.94 (ddd, J=24.6, 12.1, 5.5Hz, 2H), 0.83–0.75 (m, 1H), 0.64 (s, 3H).

[0081] MS: m / z[M+H] + 364.15

[0082] Biological activity test

[0083] I. GABA-A receptor positive regulation activity assay

[0084] Experimental methods:

[0085] The experiment used HEK293 cells transiently expressing GABA-A (α4β3δ) receptors to study the effects of compounds on GABA-A (α4β3δ) receptors and to test the half-maximal effect concentration (EC 50 The experiments were performed using a manual patch clamp system HEKAEPC 10 signal amplifier and digital conversion system to record the chloride current of GABA-A channels in whole cells. The experiments were performed at room temperature.

[0086] After the whole-cell seal is formed, the cell membrane voltage is clamped at -70mV. Recording is performed in the gap-free mode. The peak current is obtained after the cell surface is sprayed with a mixture of GABA and 3μM of the compound of the present invention in sequence from low to high concentrations. Administration of the test substance: After each concentration of GABA and 3μM of the compound of the present invention is administered 1-2 times, it is rinsed with extracellular fluid for 1 minute and then the next concentration is detected. The microelectrode pulling instrument pulls the capillary glass tube into a recording electrode. The electrode filled with intracellular fluid is loaded into the electrode head, and the microelectrode manipulator is manipulated under an inverted microscope to immerse the electrode in the extracellular fluid and record the electrode resistance (Rpip). The electrode is contacted to the cell surface, and negative pressure is applied to form a high-resistance seal (GΩ). At this time, fast capacitance compensation is performed, and negative pressure is continued to be applied to break the cell membrane and form a whole-cell recording mode. Then slow capacitance compensation is performed and experimental parameters such as membrane capacitance (Cm) and series resistance (Rs) are recorded. No leakage compensation is given.

[0087] The coverslip with cells was placed in a recording bath under an inverted microscope, and the blank control external solution and the working solution of the compound of the present invention were flowed through the recording bath in sequence from low concentration to high concentration by gravity perfusion to act on the cells, and a peristaltic pump was used to exchange the liquid during recording. At least two cells were used for each concentration and the test was repeated twice independently.

[0088] The GABA-A (α4β3δ) receptor current after the action of each concentration of GABA and the mixture of the compound of the present invention at 3 μM was normalized with the current of the mixture of saturated GABA and the compound of the present invention at 3 μM as the control. The mean, standard deviation and standard error were calculated for each concentration ratio.

[0089] Y=Bottom+(Top-Bottom) / (1+10^((LogEC 50 -X)×HillSlope))

[0090] The EC of the compounds of the present invention was calculated by the above equation: 50 The dose-dependent effect was fitted nonlinearly, where EC 50 is the half-maximal effect concentration. EC 50 The calculation and curve fitting were completed using GraphPad Prism software.

[0091] While keeping the experimental scheme and experimental parameters unchanged, the above experiment was repeated with allopregnanolone instead of the compound of the present invention, and the EC of allopregnanolone was calculated by the above equation. 50 The dose-dependent effect was fitted nonlinearly, where EC 50 is the half-maximal effect concentration. EC 50The calculation and curve fitting were completed using GraphPad Prism software.

[0092] Experimental results:

[0093] The half-maximal effect concentration (EC 50 ) value is 10.514nM, and the half-maximal effect concentration (EC 50 ) value was 8.328 nM, indicating that compound 1 of the present invention has a strong positive regulatory effect on GABA-A receptors, and its activity is close to that of allopregnanolone ( Figure 1 ).

[0094] II. In vitro metabolic stability assay

[0095] Experimental background

[0096] Cytochrome P450 oxidase (CYP450), also known as liver microsomal mixed function oxidase, is mostly located on the endoplasmic reticulum and the inner wall of mitochondria, and is involved in the metabolism of a variety of endogenous and exogenous substances such as drugs, carcinogens, steroid hormones and fatty acids. CYP450 oxidoreductase (POR) is the only electron donor for all liver microsomal enzymes, and transfers electrons to CYP450 enzymes through reduced nicotinamide adenine dinucleotide phosphate (NADPH). After obtaining electrons, CYP450 enzymes undergo redox reactions with substrates, thereby exerting metabolic activity.

[0097] Liver microsomes contain most of the phase I enzymes, the most important of which is the microsomal mixed function oxidase system with CYP450 as the main component. When conducting research with liver microsomes, if the corresponding cofactor NADPH is added, the in vitro metabolic system can be reconstructed, thereby conducting phase I metabolic stability studies through in vitro incubation methods.

[0098] Purpose

[0099] The in vitro metabolic stability of compound 1 and allopregnanolone was studied using a commercially available male SD rat liver phase I metabolic stability kit (containing liver microsomes and reagents used in the study).

[0100] Experimental reagents

[0101] Phase I metabolic stability kit (purchased from Beijing Huizhiheyuan Biotechnology Co., Ltd.), the product composition is as follows:

[0102] name Specification quantity Liquid A (20×) 1mL / vial 1 piece Liquid B (100×) 200μL / vial 1 piece Liver microsomes (20 mg / mL) 500μL / vial 1 piece Positive substrate (testosterone, 10 mM) 50μL / vial 1 piece 0.1M PBS buffer (pH 7.4) 12mL / bottle 2 bottles

[0103] The components of solution A are: 26.1mM NADP+, 66mM 6-phosphate glucose, 66mM magnesium chloride; the components of solution B are: 40U / mL 6-phosphate glucose dehydrogenase, 5mM sodium citrate;

[0104] Test substance: Compound 1 Solution: Dissolve in acetonitrile to make a 40 μg / ml solution

[0105] Allopregnanolone solution: Dissolve in acetonitrile to make a 40 μg / ml solution

[0106] Experimental methods

[0107] The positive substrate testosterone solution (10 mM) was diluted with acetonitrile to a 200 μM solution for later use; the stop solution & internal standard: testosterone-D3 was dissolved in acetonitrile to prepare a 5 ng / ml solution for later use.

[0108] 1. Experimental Group

[0109] 1) Melt the components of the kit in an ice bath and place on ice for later use;

[0110] 2) Preparation of incubation system (taking 200 μL system as an example):

[0111]

[0112] 3) Add 160 μL of pre-incubation solution B to the centrifuge tube containing 40 μL of pre-incubation solution A, pipette and swirl three times to mix, and immediately place in a 37°C water bath for incubation and timing;

[0113] 4) At the set incubation time points of 0, 5, 10, 15, 30, 45, and 60 minutes, quantitatively remove the incubation solution and add 200 μL of stop solution (precooled 4°C 5 ng / ml testosterone-D3 acetonitrile solution) to terminate the reaction (stop solution volume: incubation system volume = 1:1). Experimental samples were run in triplicate at each time point.

[0114] 2. Positive control group

[0115] In order to verify the effectiveness of the experiment, a positive control group was set up in the whole experiment to evaluate whether the incubation system was working properly. The preparation method was as follows: the test substance was replaced with the positive substrate testosterone (200 μM acetonitrile solution) in the Phase I metabolic stability kit, and the other components of the incubation system and the experimental methods were the same as the test group.

[0116] 3. Data processing

[0117] The peak area ratio of the chromatographic peak area of ​​the test substance and the peak area ratio of the testosterone-D3 internal standard was used as the calculation data. The peak area ratio of the test substance at zero time was taken as 100%, and the peak area ratio at each time point was compared with the peak area ratio at zero time to obtain the remaining percentage. Then, the natural logarithm of the substrate remaining percentage at each time point and the incubation time were linearly regressed to obtain the slope k. Thus, the in vitro metabolic half-life (T 1 / 2 ) and intrinsic clearance (CL int ), the formula is as follows:

[0118]

[0119] The test results are as follows:

[0120]

[0121] Half-life and intrinsic clearance calculation results

[0122] Compound <![CDATA[T 1 / 2 (min)]]> k <![CDATA[In vitro CL int (μL / min / mg protein) <!-- 13 -->]]> Compound 1 93.54 -0.0074 14.82 Allopregnanolone 2.17 -0.3196 639.28

[0123] The experimental results showed that the positive control testosterone was basically metabolized in 10 minutes, indicating that the incubation system worked normally. Allopregnanolone was metabolized very quickly in the liver microsomes in vitro, and was basically metabolized in 10 minutes, with a metabolic half-life of 2.17 minutes and an intrinsic clearance rate of 639.28 μL / min / mg, while the compound 1 of the present invention was metabolized more slowly, with a metabolic half-life of 93.54 minutes and an intrinsic clearance rate of 14.82 μL / min / mg.

[0124] III. Pharmacokinetic analysis of tail vein administration in mice

[0125] Experimental drugs

[0126] Allopregnanolone: ​​0.5 mg / ml aqueous solution (solubilized with 30% cyclodextrin)

[0127] Compound 1: 0.5 mg / ml aqueous solution (solubilized with 30% cyclodextrin)

[0128] Experimental animals

[0129] Female ICR mice, weighing 20-22 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0130] Experimental methods

[0131] 1. Mouse tail vein administration and treatment:

[0132] 60 female ICR mice were divided into two groups, A and B, with 30 mice in each group (5 animals at each blood collection point), and allopregnanolone and compound 1 were injected through the tail vein, respectively, with a dose of 5 mg / kg. Eyeballs were removed and blood was collected at 5min, 30min, 1h, 5h, 8h, and 12h after administration. The blood was anticoagulated with heparin and centrifuged to separate plasma and stored at -80℃ for testing; the animals at the 5min, 1h, and 8h blood collection points were killed and brain tissues were obtained and stored at -80℃ for testing.

[0133] 2. Biological sample processing

[0134] (1) Plasma samples

[0135] Accurately measure 100 μl of plasma sample, 10 μl of internal standard solution (100 ng / ml testosterone D-3 solution), and 310 μl of acetonitrile, vortex to mix, centrifuge (14000 rpm, 4°C), and take the supernatant.

[0136] (2) Brain tissue samples

[0137] Take about 100 mg of brain tissue sample, accurately weigh it, accurately add 100 μl of water and 400 μl of acetonitrile to homogenize; accurately measure 100 μl of brain tissue homogenate, 10 μl of internal standard solution (100 ng / ml testosterone D-3 solution), and 310 μl of acetonitrile, vortex mix, centrifuge (14000 rpm, 4°C), and take the supernatant.

[0138] 3. Biological sample measurement

[0139] (1) Preparation of a series of standard solutions

[0140] ① Plasma matrix standard solution

[0141] Take appropriate amount of allopregnanolone and compound 1, add acetonitrile for quantitative dilution to prepare mixed standard solutions containing 5000ng, 2000ng, 1000ng, 500ng, 200ng, 100ng, 50ng, and 20ng of allopregnanolone and compound 1 per ml. Accurately measure 100μl blank plasma sample, 10μl mixed standard solution, 10μl internal standard solution (100ng / ml testosterone D-3 solution), and 300μl acetonitrile, vortex mix, centrifuge (14000rpm, 4°C), and take the supernatant to obtain.

[0142] ②Brain tissue matrix standard solution

[0143] Take appropriate amount of allopregnanolone and compound 1, add acetonitrile to quantitatively dilute to prepare mixed standard solutions containing 5000ng, 2000ng, 1000ng, 500ng, 200ng, 100ng, and 50ng of allopregnanolone and compound 1 per 1ml. Take about 100mg of blank brain tissue sample, accurately weigh, accurately add 100μl water and 400μl acetonitrile, and homogenize with a homogenizer; accurately measure 100μl brain tissue homogenate, 10μl mixed standard solution, 10μl internal standard solution (100ng / ml testosterone D-3 solution), 300μl acetonitrile, vortex mix, centrifuge (14000rpm, 4℃) and take the supernatant to obtain.

[0144] (2) Analysis methods

[0145] The UPLC-MS / MS method was used, testosterone D-3 was used as the internal standard substance, and the standard curve method was used for analysis and determination.

[0146] UPLC parameters are as follows:

[0147]

[0148]

[0149] MS parameters

[0150]

[0151] Note: ※ indicates quantitative ions.

[0152] 4. Experimental Results

[0153]

[0154] After administration via the tail vein, both compound 1 and allopregnanolone can quickly pass through the blood-brain barrier, thus entering the brain to exert their drug effects. However, the clearance rate of compound 1 in the brain is much slower than that of allopregnanolone; the brain tissue concentrations (ng / g) of compound 1 at 5min, 1h and 8h after administration are 1138.9, 652.0 and 31.0, respectively, while the brain tissue concentrations (ng / g) of allopregnanolone at 5min, 1h and 8h after administration are 1073.2, 47.3 and 14.4, respectively; indicating that the metabolism of compound 1 in brain tissue is more stable.

[0155] IV. Pharmacokinetic analysis of tail vein and oral administration in female rats

[0156] Experimental drugs

[0157] Tail vein injection:

[0158] Allopregnanolone: ​​0.5 mg / ml aqueous solution (solubilized with 30% cyclodextrin, referred to as: A 静脉 )

[0159] Compound 1: 0.5 mg / ml aqueous solution (solubilized with 30% cyclodextrin, referred to as B 静脉 )

[0160] Oral gavage administration:

[0161] Allopregnanolone: ​​1.5 mg / ml oil solution (with sesame oil as solvent, referred to as: A 口服 )

[0162] Compound 1: 1.5 mg / ml oil solution (with sesame oil as solvent, referred to as B 口服 )

[0163] Experimental animals

[0164] Female SD rats, weighing 200-220 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0165] Experimental methods

[0166] 1. Rat tail vein administration and treatment:

[0167] Sixty female SD rats were divided into two groups (A vein and B vein), with 30 rats in each group (5 rats at each blood collection point). Drug A was injected into the tail vein. 静脉 and B 静脉 , 3 mg / kg each (about 1.2 ml of drug solution). Blood was collected from the orbital venous plexus at 5 min, 30 min, 1 h, 2 h, 4 h, and 8 h after administration. The blood was anticoagulated with heparin and centrifuged to separate the plasma, which was stored at -80 °C for testing. The animals were killed 8 h after blood collection, and brain and liver tissues were collected and stored at -80 °C for testing.

[0168] 2. Oral administration and treatment of rats:

[0169] Sixty female SD rats were divided into two groups (A oral administration and B oral administration), with 30 rats in each group (5 rats at each blood collection point). 口服 and B 口服 , 12mg / kg each (about 1.6ml of drug solution). Blood was collected from the orbital venous plexus at 5min, 30min, 1h, 2h, 4h, and 8h after administration. The blood was anticoagulated with heparin and centrifuged to separate the plasma and store it at -80℃ for testing. The animals were killed 8h after blood collection, and brain and liver tissues were collected and stored at -80℃ for testing.

[0170] 3. Sample Processing

[0171] 1) Plasma samples

[0172] Accurately measure 100 μl of plasma sample, 10 μl of internal standard solution (100 ng / ml testosterone D-3 solution), and 310 μl of acetonitrile, vortex mix, centrifuge (14000 rpm, 4°C), and take the supernatant for UPLC-MS / MS detection (the analysis method is the same as the mouse pharmacokinetic analysis experiment).

[0173] 2) Brain tissue and liver tissue samples

[0174] Take about 100 mg of brain tissue or liver tissue sample respectively, accurately weigh, accurately add 100 μl of water and 400 μl of acetonitrile for homogenization; accurately measure 100 μl of brain tissue or liver tissue homogenate, 10 μl of internal standard solution (100 ng / ml testosterone D-3 solution), 310 μl of acetonitrile, vortex mix, centrifuge (14000 rpm, 4°C), take the supernatant, and detect by UPLC-MS / MS (the analysis method is the same as the mouse pharmacokinetic analysis experiment).

[0175] 4. Experimental Results

[0176] Pharmacokinetic parameters after tail vein administration in rats

[0177] Compound Compound 1 Allopregnanolone Clearance (L / h / kg) 5.27 6.53 <![CDATA[T 1 / 2 (h)]]> 0.96 0.87 Brain / plasma ratio (8h) 6.68 No detectable levels in brain tissue or plasma

[0178] Pharmacokinetic parameters after oral administration in rats

[0179] Compound Compound 1 Allopregnanolone Clearance (L / h / kg) 0.58 / Oral bioavailability (%) ＞65 0 Brain / plasma ratio (8h) 2.95 Undetectable in brain tissue and plasma

[0180] Rats were administered allopregnanolone and compound 1 via the tail vein, respectively. 8 hours after administration, allopregnanolone was not detected in the brain tissue and plasma of rats in both groups, while the concentration of compound 1 in the brain tissue of rats in the compound 1 administration group was still high (209.1 ng / g), with a brain / plasma ratio of 6.68, indicating that the clearance rate of compound 1 in the brain is much slower than that of allopregnanolone, and its metabolism in the brain tissue is very stable.

[0181] After oral administration of allopregnanolone and compound 1 to rats, allopregnanolone was not detected in the plasma and brain tissue of the animals in the allopregnanolone group at any time point, indicating that allopregnanolone is basically ineffective for oral administration. After oral administration of compound 1, the drug concentration in plasma continued to increase, and C was still not detected at 8 hours. max , indicating that compound 1 is easily absorbed into the blood and has a low clearance rate, T max >8h, oral bioavailability>65%; compound 1 is easily absorbed into the blood and penetrates the blood-brain barrier after oral administration. The concentration in brain tissue is still very high (1636.8ng / g) 8h after administration, and the brain / plasma ratio is 2.95, indicating that compound 1 can continuously penetrate the blood-brain barrier to exert its efficacy after oral administration, and its metabolism in the blood is very stable.

[0182] In addition, the content of drugs in liver tissue after tail vein and oral administration of allopregnanolone and compound 1 was also determined. The results showed that allopregnanolone could not be detected in the liver 8 hours after allopregnanolone was administered by both administration methods; while 8 hours after oral administration of compound 1, the content of compound 1 in plasma was still very high (3703.4ng / g) (allopregnanolone (694.2ng / g) was also detected in liver tissue), and 8 hours after intravenous administration of compound 1, there was a small amount of compound 1 (192.8ng / g) in the liver, and allopregnanolone could not be detected. The results show that the metabolism of compound 1 in the liver is very stable.

[0183] V. Pharmacokinetic analysis of oral administration in male rats

[0184] Experimental drugs

[0185] Oral gavage administration:

[0186] Allopregnanolone: ​​1.5 mg / ml oil solution (with sesame oil as solvent, referred to as: A 口服 )

[0187] Compound 1: 1.5 mg / ml oil solution (with sesame oil as solvent, referred to as B 口服 )

[0188] Experimental animals

[0189] Male SD rats, weighing 230-250 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd.

[0190] Experimental methods

[0191] 1. Oral administration and treatment of rats:

[0192] A total of 168 male SD rats were divided into two groups (A 口服 and B 口服 ), 84 mice in each group (7 mice at each blood collection point). Drug A was administered orally by gavage 口服 and B 口服 , 12 mg / kg each. Blood was collected from the orbital venous plexus at 30 min, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 8 h, 10 h, 12 h, 18 h, and 24 h after administration. The blood was anticoagulated with heparin and centrifuged to separate the plasma, which was then stored at -80 °C for testing.

[0193] 2. Sample Processing

[0194] Accurately measure 100 μl of plasma sample, 10 μl of internal standard solution (100 ng / ml testosterone D-3 solution), and 310 μl of acetonitrile, vortex mix, centrifuge (14000 rpm, 4°C), and take the supernatant for UPLC-MS / MS detection (the analysis method is the same as the mouse pharmacokinetic analysis experiment).

[0195] 3. Experimental results

[0196]

[0197]

[0198] After oral administration of allopregnanolone and compound 1 to male rats, allopregnanolone was not detected in the plasma of the animals in the allopregnanolone group at any time point, indicating that allopregnanolone was basically not absorbed into the blood after oral administration. However, after oral administration of compound 1, the drug concentration in plasma continued to increase, reaching C max , indicating that compound 1 is easily absorbed into the blood and has a low clearance rate, T max About 10h, half-life is 4.5h.

[0199] The references mentioned herein are all incorporated herein by reference. It should be understood that the technical solutions of the present invention can be changed and modified without departing from the spirit and scope of the present disclosure.

Claims

1. A compound having a structure represented by the following formula (IV), or its deuterated substance and pharmaceutically acceptable salt, wherein: R1 is selected from hydrogen or C1-C 12 alkyl; R2 is selected from R-(C=O)-, wherein R is selected from hydrogen or C1-C 12 alkyl.

2. The compound according to claim 1 or its deuterated substance and pharmaceutically acceptable salt, wherein: R1 is selected from hydrogen or C1-C6 alkyl; R2 is selected from R-(C=O)-, wherein R is selected from hydrogen or C1-C 12 alkyl.

3. The compound according to claim 1 or its deuterated substance and pharmaceutically acceptable salt, wherein: R1 is selected from hydrogen or C1-C6 alkyl; R2 is selected from R-(C=O)-, wherein R is selected from hydrogen or C1-C6 alkyl.

4. The compound according to claim 1 or its deuterated substance and pharmaceutically acceptable salt, wherein: R1 is selected from hydrogen, methyl, ethyl, n-propyl or isopropyl; R2 is selected from R-(C=O)-, wherein R is selected from hydrogen, methyl, ethyl, n-propyl or isopropyl.

5. The compound according to claim 1, which is or a deuterated substance or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 or a deuterated substance thereof and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

7. Use of a compound according to any one of claims 1 to 5 in the preparation of a medicament for the treatment and / or prevention of GABA-A receptor-mediated diseases.

8. The method of claim 7, wherein the GABA-A receptor-mediated disease is selected from postpartum depression, depression, major depressive disorder, bipolar disorder, mood disorders, anxiety, post-traumatic stress disorder (PTSD), premenstrual dysphoric disorder (PMDD), premenstrual syndrome, generalized anxiety disorder, seasonal affective disorder (SAD), social anxiety disorder, memory loss, stress intolerance, Niemann-Pick disease type C or related neurological or physical symptoms, epilepsy, essential tremor, epileptiform disorders, NMDA hypofunction, migraine, status epilepticus, sleep disorders, fragile X syndrome, 5α-reductase inhibitor-induced depression, PCDH19 female pediatric epilepsy, sexual dysfunction, cognitive disorders, Parkinson's disease or Alzheimer's disease.

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