WO2021027792A1 - Vmat2抑制剂及其制备方法和应用 - Google Patents

Vmat2抑制剂及其制备方法和应用 Download PDF

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WO2021027792A1
WO2021027792A1 PCT/CN2020/108314 CN2020108314W WO2021027792A1 WO 2021027792 A1 WO2021027792 A1 WO 2021027792A1 CN 2020108314 W CN2020108314 W CN 2020108314W WO 2021027792 A1 WO2021027792 A1 WO 2021027792A1
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compound
alkyl
pharmaceutically acceptable
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English (en)
French (fr)
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田京伟
张睿
叶亮
于大伟
杜广营
刘宗亮
邹方霞
崔博
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Shandong Luye Pharmaceutical Co Ltd
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Shandong Luye Pharmaceutical Co Ltd
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Priority to CN202210968124.3A priority Critical patent/CN115260186B/zh
Priority to JP2022506161A priority patent/JP7702384B2/ja
Priority to BR112022002107A priority patent/BR112022002107A2/pt
Priority to CA3148302A priority patent/CA3148302A1/en
Priority to CN202080052104.0A priority patent/CN114302883B/zh
Priority to KR1020227005179A priority patent/KR20220035204A/ko
Application filed by Shandong Luye Pharmaceutical Co Ltd filed Critical Shandong Luye Pharmaceutical Co Ltd
Priority to AU2020327501A priority patent/AU2020327501B2/en
Priority to US17/634,977 priority patent/US12565495B2/en
Priority to EP20852655.8A priority patent/EP4015517A4/en
Publication of WO2021027792A1 publication Critical patent/WO2021027792A1/zh
Anticipated expiration legal-status Critical
Priority to US19/414,124 priority patent/US20260091029A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/14—Drugs for disorders of the nervous system for treating abnormal movements, e.g. chorea, dyskinesia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C07D455/03—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
    • C07D455/04—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing a quinolizine ring system condensed with only one six-membered carbocyclic ring, e.g. julolidine
    • C07D455/06—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing a quinolizine ring system condensed with only one six-membered carbocyclic ring, e.g. julolidine containing benzo [a] quinolizine ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/13—Crystalline forms, e.g. polymorphs

Definitions

  • the present invention relates to a class of compounds as VMAT2 inhibitors or their stereoisomers or pharmaceutically acceptable salts thereof, and their application in the field of diseases related to VMAT2.
  • Tardive dyskinesia also known as tardive hyperactivity
  • TD Tardive dyskinesia
  • the disease is more common in patients who take large doses of antipsychotic drugs for a long time.
  • the main clinical feature is involuntary, rhythmic repetitive and stereotyped movement, often involving the jaw, lips, and tongue.
  • Other drugs can also cause tardive dyskinesias, such as Parkinson's drugs (levodopa) and so on.
  • VMAT2 Vesicular monoamine transporter 2
  • DA dopamine
  • VMAT2 inhibitors can antagonize the reuptake function of VMAT2, so that dopamine cannot be re-uptaken into vesicles by VMAT2, but is metabolized by enzymes in the cytoplasm, resulting in a reduction in dopamine release in the synaptic cleft, thereby realizing the treatment of tardive dyskinesia purpose.
  • Tetrabenazine (TBZ, alias tetrabenazine) is the first VMAT2 selective inhibitor to be marketed.
  • the metabolite trans(2,3)-dihydrotetrabenazine (DHTBZ) in the body also has VMAT2 selection Sexual inhibitory activity.
  • DHTBZ metabolite trans(2,3)-dihydrotetrabenazine
  • Valbenazine is esterified from the metabolite DHTBZ of tetrabenazine, which is better than tetrabenazine.
  • Ozine has a long half-life and does not require frequent administration. Its curative effect is clear, and its safety and tolerance are good.
  • Tetrabenazine has the problem of short half-life and the need for multiple administrations, and there are many types of tetrabenazine metabolites, which cause serious adverse reactions, and there are black box adverse reactions warnings of depression and suicidal tendency. There are many people with tardive dyskinesia, but there are fewer drugs on the market. Therefore, there is still a need for more active VMAT2 inhibitors in this field to meet a wide range of clinical needs.
  • the present invention provides a compound represented by formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof.
  • R is selected from OH, H or
  • R is O
  • R 1 is selected from hydrogen, methyl or ethyl
  • R 2 3 is selected from unsubstituted or substituted with 1, 2 or 3 R C 2-10 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl -C 1-6 alkyl group, 3 ⁇ 6-membered heterocycloalkyl-C 1-3 alkyl, C 2-6 alkenyl or C 1-6 heteroalkyl;
  • R 3 is selected from F, Cl, Br, OH, SH or NH 2 .
  • R 2 of the compound of formula (I) is selected from unsubstituted C 2-10 alkyl, C 3-6 cycloalkyl-C 1-6 alkyl, or substituted by 1, 2 or 3
  • R 3 is a substituted C 2-10 alkyl; preferably unsubstituted or substituted with 2-3 R 3 is C 2-5 alkyl; R 3 is F, the other variables are as defined in the present invention.
  • R 2 of the compound of formula (I) is selected from ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl, trifluoropropyl, trifluorobutyl Group, trifluoropentyl, bisfluoroethyl or cyclopropanemethylene; preferably trifluoroethyl or cyclopropanemethylene, other variables are as defined in the present invention.
  • R 1 in the compound of formula (I) is methyl; "---" is a single bond, and R is selected from OH or Or "---" is a double bond, R is O; other variables are as defined in the present invention.
  • the heteroatoms in the 3--6 membered heterocycloalkyl or C 1-6 heteroalkyl of the compound of formula (I) are O, S or N; the number of heteroatoms is 1-6 , Preferably 1 O; the number of C atoms of the C 1-6 heteroalkyl group is 2-6 or 3-6 or 2-5 or 3-5 or 4-6 or 4-5.
  • the present invention also provides a compound of the structure represented by formula (II) or a stereoisomer or a pharmaceutically acceptable salt thereof.
  • R 2 is selected from C 2-10 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl-C 1-3 alkyl that is unsubstituted or substituted by 1, 2 or 3 R 3 , C 2-6 alkenyl, C 1-6 heteroalkyl; R 3 is selected from F, Cl, Br, OH, NH 2 .
  • R 2 of the compound of formula (II) is selected from unsubstituted C 2-10 alkyl, C 3-6 cycloalkyl-C 1-6 alkyl, or is substituted by 1, 2 or 3
  • R 3 is a substituted C 2-10 alkyl; preferably is unsubstituted or substituted with 2-3
  • R 3 is C 2-5 alkyl;
  • R 3 is F.
  • R 2 of the compound of formula (II) is selected from ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl, trifluoropropyl, trifluorobutyl Group, trifluoropentyl, bisfluoroethyl or cyclopropanemethylene; preferably trifluoroethyl or cyclopropanemethylene.
  • the heteroatom in the C 1-6 heteroalkyl group of the compound of formula (II) is O, S or N; the number of heteroatoms is 1-6, preferably 1 O.
  • the present invention also provides a compound of the structure represented by formula (III) or a stereoisomer or a pharmaceutically acceptable salt thereof,
  • R 2 is selected from C 2-10 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl-C 1-3 alkyl that is unsubstituted or substituted by 1, 2 or 3 R 3 , C 2-6 alkenyl, C 1-6 heteroalkyl; R 3 is selected from F, Cl, Br, OH, NH 2 .
  • R 2 of the compound of formula (III) is selected from unsubstituted C 2-10 alkyl, C 3-6 cycloalkyl-C 1-6 alkyl, or substituted by 1, 2 or 3
  • R 3 is a substituted C 2-10 alkyl; preferably is unsubstituted or substituted with 2-3
  • R 3 is C 2-5 alkyl;
  • R 3 is F.
  • R 2 of the compound of formula (III) is selected from ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl, trifluoropropyl, trifluorobutyl Group, trifluoropentyl, bisfluoroethyl or cyclopropanemethylene; preferably trifluoroethyl or cyclopropanemethylene.
  • the heteroatom in the C 1-6 heteroalkyl group of the compound of formula (III) is O, S or N; the number of heteroatoms is 1-6, preferably 1 O.
  • the present invention provides a compound of the structure represented by formula (IV) or a stereoisomer or a pharmaceutically acceptable salt thereof,
  • R 2 is selected from C 2-10 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl-C 1-3 alkyl that is unsubstituted or substituted by 1, 2 or 3 R 3 , C 2-6 alkenyl, C 1-6 heteroalkyl; R 3 is selected from F, Cl, Br, OH, NH 2 .
  • R 2 of the compound of formula (IV) is selected from unsubstituted C 2-10 alkyl, C 3-6 cycloalkyl-C 1-6 alkyl, or substituted by 1, 2 or 3
  • R 3 is a substituted C 2-10 alkyl; preferably is unsubstituted or substituted with 2-3
  • R 3 is C 2-5 alkyl;
  • R 3 is F.
  • R 2 of the compound of formula (IV) is selected from ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl, trifluoropropyl, trifluorobutyl Group, trifluoropentyl, bisfluoroethyl or cyclopropanemethylene; preferably trifluoroethyl or cyclopropanemethylene.
  • the heteroatom in the C 1-6 heteroalkyl group of the compound of formula (IV) is O, S or N; the number of heteroatoms is 1-6, preferably 1 O.
  • valine esters and pharmaceutically acceptable salts of the following structural compounds are also provided:
  • the present invention also provides the p-toluene sulfonate of any one of the above compounds, preferably the following compounds:
  • the present invention also provides five crystal forms of compound 11-P4S.
  • the X-ray powder diffraction pattern of 11-P4S crystal form A obtained by Cu-Ka radiation contains the following characteristic peaks measured by 2 ⁇ reflection angle: 6.33 ⁇ 0.2°, 10.87 ⁇ 0.2°, 18.89 ⁇ 0.2° .
  • the X-ray powder diffraction pattern of 11-P4S crystal form A obtained by Cu-Ka radiation contains the following characteristic peaks measured by the 2 ⁇ reflection angle: 6.33 ⁇ 0.2°, 10.87 ⁇ 0.2°, 16.61 ⁇ 0.2° , 18.89 ⁇ 0.2°, 19.27 ⁇ 0.2°, 22.19 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of 11-P4S crystal form A obtained by Cu-Ka radiation contains the following characteristic peaks measured by 2 ⁇ reflection angle: 6.33 ⁇ 0.2°, 10.87 ⁇ 0.2°, 13.77 ⁇ 0.2° , 16.61 ⁇ 0.2°, 18.20 ⁇ 0.2°, 18.89 ⁇ 0.2°, 19.27 ⁇ 0.2°, 20.05 ⁇ 0.2°, 22.19 ⁇ 0.2°, 24.60 ⁇ 0.2°, 24.77 ⁇ 0.2°.
  • 11-P4S crystal form A has an X-ray powder diffraction pattern as shown in Fig. 2-1 through Cu-Ka radiation.
  • the X-ray powder diffraction pattern of 11-P4S crystal form B obtained by Cu-Ka radiation contains the following characteristic peaks measured by the 2 ⁇ reflection angle: 6.32 ⁇ 0.2°, 5.42 ⁇ 0.2°, 10.85 ⁇ 0.2° .
  • the X-ray powder diffraction pattern of 11-P4S crystal form B obtained by Cu-Ka radiation contains the following characteristic peaks measured by the 2 ⁇ reflection angle: 6.32 ⁇ 0.2°, 5.42 ⁇ 0.2°, 10.85 ⁇ 0.2° , 16.60 ⁇ 0.2°, 18.88 ⁇ 0.2°, 22.02 ⁇ 0.2°.
  • the 11-P4S crystal form B has an X-ray powder diffraction pattern substantially as shown in FIG. 3-1 through Cu-Ka radiation.
  • the X-ray powder diffraction pattern of 11-P4S crystal form C obtained by Cu-Ka radiation contains the following characteristic peaks measured by the 2 ⁇ reflection angle: 5.81 ⁇ 0.2°, 6.33 ⁇ 0.2°, 12.86 ⁇ 0.2° .
  • the X-ray powder diffraction pattern of 11-P4S crystal form C obtained by Cu-Ka radiation includes the following characteristic peaks measured by 2 ⁇ reflection angle: 5.81 ⁇ 0.2°, 6.33 ⁇ 0.2°, 7.99 ⁇ 0.2° , 12.86 ⁇ 0.2°, 19.09 ⁇ 0.2°, 23.17 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of 11-P4S crystal form C obtained by Cu-Ka radiation includes the following characteristic peaks measured by 2 ⁇ reflection angle: 5.81 ⁇ 0.2°, 6.33 ⁇ 0.2°, 7.99 ⁇ 0.2° , 10.31 ⁇ 0.2°, 11.63 ⁇ 0.2°, 12.86 ⁇ 0.2°, 18.16 ⁇ 0.2°, 19.09 ⁇ 0.2°, 23.17 ⁇ 0.2°, 24.00 ⁇ 0.2°, 27.32 ⁇ 0.2°.
  • the 11-P4S crystal form C has an X-ray powder diffraction pattern as shown in Fig. 4-1 through Cu-Ka radiation.
  • the X-ray powder diffraction pattern of 11-P4S crystal form D obtained by Cu-Ka radiation includes the following characteristic peaks measured by 2 ⁇ reflection angle: 6.02 ⁇ 0.2°, 23.91 ⁇ 0.2°.
  • the X-ray powder diffraction pattern of 11-P4S crystal form D obtained by Cu-Ka radiation contains the following characteristic peaks measured by 2 ⁇ reflection angle: 5.31 ⁇ 0.2°, 6.02 ⁇ 0.2°, 18.88 ⁇ 0.2° , 22.12 ⁇ 0.2°, 23.91 ⁇ 0.2°.
  • the 11-P4S crystal form D has an X-ray powder diffraction pattern as shown in Fig. 5-1 through Cu-Ka radiation.
  • the X-ray powder diffraction pattern of 11-P4S crystal form E obtained by Cu-Ka radiation includes the following characteristic peaks measured by 2 ⁇ reflection angle: 6.06 ⁇ 0.2°, 18.32 ⁇ 0.2°, 30.79 ⁇ 0.2°
  • the 11-P4S crystal form E has an X-ray powder diffraction pattern as shown in Fig. 6-1 through Cu-Ka radiation.
  • the present invention also provides a crystal form of 19P2 p-toluene sulfonate (referred to as 19P2S for short), which belongs to the P212121 space group of the orthorhombic system.
  • 19P2S a crystal form of 19P2 p-toluene sulfonate
  • the present invention also provides a pharmaceutical composition, comprising any of the above-mentioned compounds or their stereoisomers or pharmaceutically acceptable salts thereof, or the crystalline form of any of the above-mentioned compounds and a pharmaceutically acceptable carrier .
  • the pharmaceutical composition can be prepared into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral liquids, granules, injections, or various sustained and controlled release formulations.
  • the pharmaceutical composition can be administered orally or parenterally (such as intravenously, subcutaneously or topically). The dosage can be adjusted appropriately according to the patient's age, sex and type of disease. Generally, the daily dosage is about 10-100 mg/day.
  • the present invention also provides any of the above-mentioned compounds or their stereoisomers or pharmaceutically acceptable salts thereof, or the crystalline form of any of the above-mentioned compounds, or pharmaceutical compositions for preparing medicines for the treatment of diseases related to VMAT2 Applications.
  • the present invention also provides the use of the above-mentioned compound or its stereoisomer or its pharmaceutically acceptable salt, or the crystalline form of any one of the above-mentioned compounds, or the pharmaceutical composition in the preparation of a medicament for the treatment of hyperkinetic disorders ;
  • the hyperkinetic disorder includes Huntington's disease, tardive dyskinesia, Tourette syndrome or convulsions.
  • the present invention also provides a preparation method of the compound of formula (II), which comprises the following preparation steps:
  • R 2 is the same as defined in the compound of formula (II) above, preferably R 2 is ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl , Trifluoropropyl, trifluorobutyl, trifluoropentyl, difluoroethyl or cyclopropanemethylene; more preferably trifluoroethyl or cyclopropanemethylene.
  • the present invention also provides a method for preparing the compound of formula (III), which comprises the following steps:
  • R 2 is the same as defined in the compound of formula (III) above, preferably R 2 is ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl Group, trifluoropropyl, trifluorobutyl, trifluoropentyl, difluoroethyl or cyclopropanemethylene; more preferably trifluoroethyl or cyclopropanemethylene.
  • the present invention also provides a method for preparing the compound of formula (IV), which comprises the following steps:
  • R 2 has the same definition as in the compound of formula (IV) above, preferably R 2 is ethyl, propyl, isobutyl, monofluorobutyl, monofluoropentyl, trifluoroethyl, trifluoropropyl, Trifluorobutyl, trifluoropentyl, difluoroethyl or cyclopropanemethylene; more preferably trifluoroethyl or cyclopropanemethylene.
  • the P group is an amino protecting group, preferably benzyloxycarbonyl (Cbz), tert-butoxycarbonyl (Boc), fluorenylmethyloxycarbonyl (Fmoc), allyloxycarbonyl (Alloc), trimethylsilylethoxycarbonyl ( Teoc), etc.
  • the protective group can be removed by using methods commonly used in the art. For details, see Protective Groups in Organic Synthesis, Third Edition, Author(s): Theodora W. Greene Ph.D., Chapter 7.
  • the present invention also provides a method for preparing stereoisomers of the compound of formula (I), which are specifically as follows:
  • the R is -OH or Other variables are the same as those defined in the compound of formula (I) above; the preparation method is to use a chiral chromatography column to resolve the compound of formula (I), and the chiral chromatography column is preferably a Daicel CHIRALPAKAD-H chromatography column.
  • the compound provided by the present invention has any one or more of the following advantages: strong affinity with VMAT2, higher exposure in the body, high concentration in the brain, long half-life, strong drug effect and the like.
  • pharmaceutically acceptable refers to compounds, materials, compositions and/or dosage forms that are within the scope of reliable medical judgment and are suitable for use in contact with human and animal tissues. Without excessive toxicity, irritation, allergic reaction or other problems or complications, it is commensurate with a reasonable benefit/risk ratio.
  • pharmaceutically acceptable salt refers to a salt of the compound of the present invention, which is prepared from a compound with specific substituents discovered in the present invention and a relatively non-toxic acid or base.
  • the base addition salt can be obtained by contacting the neutral form of the compound with a sufficient amount of base in a pure solution or a suitable inert solvent.
  • Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts.
  • the acid addition salt can be obtained by contacting the neutral form of the compound with a sufficient amount of acid in a pure solution or a suitable inert solvent.
  • examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, or organic acid salts. Certain specific compounds of the present invention contain basic and acidic functional groups, so they can be converted into any base or acid addition salt.
  • the pharmaceutically acceptable salt of the present invention can be synthesized from the parent compound containing acid or base by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of appropriate base or acid in water or an organic solvent or a mixture of both.
  • the compounds of the present invention may exist in specific geometric or stereoisomeric forms.
  • the present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers Conformers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomers or diastereomeric enriched mixtures, all of these mixtures belong to Within the scope of the present invention.
  • Additional asymmetric carbon atoms may be present in substituents such as alkyl. All these isomers and their mixtures are included in the scope of the present invention.
  • enantiomers refers to stereoisomers that are mirror images of each other.
  • diastereomer refers to a stereoisomer in which a molecule has two or more chiral centers and the relationship between the molecules is not mirror images.
  • wedge-shaped solid line keys and wedge-shaped dashed key Represents the absolute configuration of a three-dimensional center.
  • optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If you want to obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with chiral auxiliary agents, in which the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide pure The desired enantiomer.
  • the molecule when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), it forms a diastereomeric salt with a suitable optically active acid or base, and then passes through a conventional method known in the art The diastereoisomers are resolved, and then the pure enantiomers are recovered.
  • the separation of enantiomers and diastereomers is usually accomplished through the use of chromatography, which employs a chiral stationary phase and is optionally combined with chemical derivatization (for example, the formation of amino groups from amines). Formate).
  • the compounds of the present invention may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound.
  • compounds can be labeled with radioisotopes, such as tritium (3H), iodine-125 (125I), or C-14 (14C).
  • deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have reduced toxic side effects and increased drug stability. , Enhance the efficacy, extend the biological half-life of drugs and other advantages. All changes in the isotopic composition of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.
  • Figures 2-1, 2-2, and 2-3 XRPD, TGA/DSC, 1 H NMR of compound 11-P4S crystal form A, respectively
  • Figure 3-1, 3-2, 3-3 XRPD pattern, TGA/DSC pattern, 1 H NMR pattern of compound 11-P4S crystal form B, respectively
  • Figure 4-1, 4-2, 4-3 XRPD pattern, TGA/DSC pattern, 1 H NMR pattern of compound 11-P4S crystal form C, respectively
  • Figure 5-1, 5-2, and 5-3 XRPD, TGA/DSC, 1 H NMR of compound 11-P4S crystal form D, respectively
  • Figure 6-1, 6-2, 6-3 XRPD pattern, TGA/DSC pattern, 1 H NMR pattern of compound 11-P4S crystal form E, respectively
  • Figure 8 The ellipsoid diagram of the molecular structure of compound 19P2S
  • Figure 9 Plasma concentration-time curves of compound 11, compound 16, control example 44 compound and DHTBZ after intragastric administration to male SD rats
  • Figure 10 After intragastric administration of VBZ, compound 21, compound 22, compound 23 and control compound 45 to SD rats, the distribution of the original compound in the brain tissue
  • FIG 11 Distribution of drug tissues in brain tissue of active metabolites (DHTBZ, compound 11, compound 12, compound 13, and comparative example 44 compound)
  • Figure 12 After intragastric administration of VBZ, compound 21, compound 22, compound 23 and control compound 45 to SD rats, the original compound brain blood ratio (B/P ratio);
  • FIG 13 Active metabolites (DHTBZ, compound 11, compound 12, compound 13, and comparative example 44 compound) cerebral blood ratio (B/P ratio)
  • Step 1 Dissolve reactant I (50 mg, 0.28 mmol) in DMF (1 mL), add K 2 CO 3 (77 mg, 0.56 mmol), and stir for 0.5 h at room temperature. Trifluoroiodoethane (76mg, 0.36mmol) was added, and the reaction was stirred at 80°C overnight. TLC detected that the reaction was complete. The reaction was quenched by adding water, and a solid was precipitated, stirred for 1 h, filtered with suction, and the filter cake was washed twice with water. The filter cake was collected and dried to obtain Intermediate 1, 0.07 g of light yellow solid.
  • Step 2 Dissolve Intermediate 1 (0.07g, 0.270mmol) in a mixed solution of 1mL of ethanol and 1mL of water, add 3-dimethylamino-5-methyl-2-hexanone (0.06g, 0.324mmol), Benzyltriethylammonium chloride (0.02g, 0.081mmol) was heated to 95°C and reacted for 18h.
  • Step 1 Dissolve reactant I (50mg, 0.282mmol) in a mixed solution of 1mL ethanol and 1mL water, add 3-dimethylamino-5-methyl-2-hexanone (0.06g, 0.338mmol), benzyl Triethylammonium chloride (0.02g, 0.085mmol) was heated to 95°C and reacted for 18h.
  • compound 6 (610 mg, 1.58 mmol) was dissolved in 10 mL THF, sodium borohydride (120 mg, 3.17 mmol) and 10 mL MeOH were added under ice bath, and the reaction was carried out for 2 h in ice bath.
  • Examples 12-20 The synthesis of Examples 12-20 was carried out in a similar manner to that of Example 11.
  • the reaction system was poured into a mixed solution of water (3.5L) and saturated brine (700mL), extracted with ethyl acetate (700ml) four times, and the ethyl acetate phases were combined.
  • the ethyl acetate phase was washed once with water (700 mL) and saturated sodium chloride solution (700 mL).
  • the ethyl acetate phase was dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the ethyl acetate phase was concentrated under reduced pressure.
  • the solid was washed with water (300 mL) each time for a total of three washings. After drying, 39 g of light yellow solid was obtained.
  • the solid was dissolved in 80°C ethyl acetate (300 mL), then petroleum ether (100 mL) was added, and the temperature was naturally cooled for 4 hours, and a white solid was precipitated.
  • the solids were collected and washed with a mixed solution of ethyl acetate (50 mL) and petroleum ether (50 mL) each time for a total of three washings. After drying, compound 11 (23.07 g, white solid) was obtained, yield: 59.6%.
  • Peak number keep time Peak area Relative peak area% 1 4.994 5152535 48.604 2 5.601 114788 1.083 3 6.109 5177576 48.840 4 6.707 156247 1.474
  • Peak number keep time Peak area Relative peak area% 1 1.920 502928 3.811 2 4.939 12433977 94.208 3 6.139 29066 0.220 4 6.440 232459 1.761
  • Peak number keep time Peak area Relative peak area% 1 1.919 512872 5.985 2 4.999 150632 1.758 3 6.056 7906161 92.258
  • Step 1 Compound 11-P4 becomes mono-p-toluenesulfonate:
  • 11-P4 (0.20g, 0.52mmol) was dissolved in ethyl acetate (5ml), and the ethyl acetate solution of p-toluenesulfonic acid monohydrate (0.12g, 0.62mmol) was added dropwise to precipitate a white solid.
  • Step 2 Single crystal cultivation method of compound 11-P4S crystal form A
  • Step 3 XRSD test of compound 11-P4S crystal form A:
  • Light source Cu target micro-focus spot light source; 50W;
  • SHELXT (Sheldrick, GM2015. ActaCryst. A71, 3-8) was used for structural analysis, and SHELXL (against F 2 ) (Sheldrick, GM2015. ActaCryst. C71, 3-8) was used for structural refinement.
  • Table 9 List of single crystal diffraction data of compound 11-P4S crystal form A
  • Table 10 Atomic coordinates (x 10 ⁇ 4) and equivalent isotropic displacement parameters ((A ⁇ 2 x 10 ⁇ 3)
  • Compound 11-P4S crystal form A is a colorless block (0.20 ⁇ 0.10 ⁇ 0.10mm 3 ), belonging to the P21212 space group of the orthorhombic system.
  • the ellipsoid diagram of 11-P4S molecular structure is shown in Figure 1.
  • the compound structure is:
  • the XPRD pattern is shown in Figure 2-1, and the analysis data of the pattern is shown in Table 12.
  • the XPRD spectrum is shown in Figure 3-1, and the spectrum analysis data is shown in Table 14.
  • the 1 H NMR spectrum is shown in Figure 3-3, which shows that the molar ratio of p-toluenesulfonic acid to free base of the sample is 1.0:1.0, and the molar ratio of THF to free base is 0.5, corresponding to a weight loss of 6.5%, and no methanol residue is detected.
  • the XPRD pattern is shown in Figure 4-1, and the analysis data of the pattern is shown in Table 15.
  • the 1 H NMR chart is shown in Figure 4-3, which shows that the molar ratio of p-toluenesulfonic acid to free base in the sample is 1.0:1.0, the molar ratio of DCM to free base is 0.2, the solvent mass fraction is 3.1%, and no methanol residue is detected. .
  • the XPRD pattern is shown in Figure 5-1, and the pattern analysis data is shown in Table 16.
  • the XPRD pattern is shown in Figure 6-1, and the pattern analysis data is shown in Table 17.
  • the 1 H NMR chart is shown in Figure 6-3, which shows that the molar ratio of p-toluenesulfonic acid to free base of the sample is 1.0:1.0, and no MTBE residue is detected.
  • Step 1 Compound 11-P3 becomes mono-p-toluenesulfonate:
  • 11-P3 (0.20g, 0.52mmol) was dissolved in ethyl acetate (5ml), and the ethyl acetate solution of p-toluenesulfonic acid monohydrate (0.12g, 0.62mmol) was added dropwise to precipitate a white solid.
  • Step 2 Single crystal cultivation method of compound 11-P3S:
  • Step 3 Single crystal XRSD test of 11-P3S:
  • Table 18 List of single crystal diffraction data of compound 11-P3S
  • Table 19 Atomic coordinates (x 10 ⁇ 4) and equivalent isotropic displacement parameters (A ⁇ 2 x 10 ⁇ 3)
  • the crystal form of compound 11-P3S is a colorless block (0.20 ⁇ 0.20 ⁇ 0.10mm 3 ) and belongs to the P21212 space group of the orthorhombic system.
  • the ellipsoid diagram of the molecular structure of compound 11-P3S is shown in Figure 7.
  • the compound structure is:
  • Boc-L-valine 857.6mg, 3.95mmol
  • 4-dimethylaminopyridine 386.9mg, 3.16mmol
  • compound 11-P4 1.02g, 2.63mmol
  • Dicyclohexylcarbodiimide 83.7 mg, 3.95 mmol was added at once, the temperature was increased naturally, and the reaction was carried out for 18 hours.
  • Peak number keep time Peak area Relative peak area% 1 3.481 246012 14.159 2 5.397 1225940 70.559 3 12.004 265519 15.282
  • Step 1 Compound 19P2 becomes mono-p-toluenesulfonate:
  • Step 3 Single crystal XRSD test of 19P2S:
  • Table 23 Atomic coordinates (x 10 ⁇ 4) and equivalent isotropic displacement parameters ((A ⁇ 2 x 10 ⁇ 3)
  • the compound 19P2S crystal is a colorless block (0.30 ⁇ 0.10 ⁇ 0.04mm3) and belongs to the P212121 space group of the orthorhombic system.
  • Boc-L-valine (260mg, 1.2mmol) in 10mL of dichloromethane, add dicyclohexylcarbodiimide (309mg, 1.5mmol), 4-dimethylaminopyridine (12.2 mg, 0.1mmol), and then add 3-isobutyl-10-methoxy-9-cyclopropylmethyl-2,3,4,6,7,11b-hexahydro-1H-pyridine [2,1- a] Isoquinolin-2-ol (359mg, 1.0mmol), warm to room temperature and stir to react overnight.
  • Step 1 Fragment 2 synthesis
  • Step 1 Fragment 3 synthesis
  • Comparative Examples 43-44 refers to the method of Comparative Example 42, which was prepared by reducing ethanol and sodium borohydride solution.
  • the IC 50 and Ki value of each compound binding to rat VMAT2 were measured, and the affinity of the compound to VMAT2 was evaluated.
  • DHTBZ Jiangsu Weikeer Pharmaceutical Technology Co., Ltd., batch number 67-25-1521-59C
  • TBZ is used as a raw material and prepared according to the Reaction Scheme 1 of WO2008058261
  • VBZ Prepare according to the following method: Dissolve VBZ xylene sulfonate (0.5g, 0.65mmol) in 10mL of water, adjust the pH to about 8 with saturated NaHCO3 solution, extract with EA (20mL*3), combine the organic phases. After drying with sodium sulfate and concentrating, VBZ 0.26g white solid is obtained.
  • a male Wistar rat weighing 175 ⁇ 25g was selected, and the whole brain (excluding the cerebellum) was surgically isolated, placed in 20 ml of pre-cooled 0.32M sucrose solution, and homogenized with a Teflon pestle homogenizer. The homogenate was centrifuged at 1000g for 12min at 4°C; the supernatant was aspirated and the supernatant was centrifuged at 22,000g for 10min at 4°C, the supernatant was discarded, and the resulting pellet was incubated in 18mL of ice-cold MilliQ water (Millipore Corporation, Billerica, MA) for 5 minutes , Perform osmotic shock to break the cell membrane.
  • ice-cold MilliQ water MilliQ water
  • vesicle suspension containing 32 ⁇ g protein
  • 10nM [3H]Dihydrotetrabenazine DHTBZ
  • 50 ⁇ L of the test compound-containing solution inhibitor concentration range of 1nM-1000nM, or other desired concentrations
  • the non-specific ligand Ro4-1284 (10 ⁇ M) was used to determine the prediction of non-specific binding
  • Tetrabenazine (TBZ), DHTBZ, VBZ or DHTBZ racemates with clear pharmacological characteristics were used as positive controls and used to Comparison of compound activity.
  • Table 27 The binding inhibition rate data of some example compounds of the present invention
  • the test results show that compared with the compounds of TBZ, DHTBZ, DHTBZ-X, VBZ and the control example, the compound provided by the present invention has a stronger affinity with VMAT.
  • the 10-position is methyl
  • the 9-position is ethyl, cyclopropylmethylene and 4-fluorobutyl
  • the binding inhibition activity is the best
  • the 10-position is propyl, butyl and other long substituents
  • the binding inhibition The activity is significantly reduced, even close to no activity.
  • the 9th position is a methyl group and the 10th position is a long substituent such as ethyl, propyl, and cyclopropylmethylene
  • the activity is significantly reduced or even no activity. That is, the inventors found that the 9-position and 10-position substituents have a significant difference in activity.
  • Frozen vesicle suspension The vesicle suspension was extracted from the striatum of SD rats. The fresh rat striatum was added to 28 mL of 0.32M sucrose solution in an ice bath, and homogenized with a homogenizer for 10 times, each for 10 seconds. Centrifuge the homogenate at 2000g for 10min at 4°C. Aspirate the supernatant and continue to centrifuge at 10000g for 30 minutes at 4°C. Separate the precipitate, resuspend in 4mL 0.32M sucrose solution, and add 14mL ice bath MilliQ water to make osmotic shock. After 1 min, add 1.8mL 0.25M HEPES buffer and 1.8mL 1M potassium tartrate solution.
  • 10mL detection buffer 25mM HEPES, 100mM potassium tartrate, 50 ⁇ M EGTA, 100 ⁇ M EDTA, 20mM MgCl2, 2mM ATP, pH7.4
  • Assay buffer 25mM HEPES, 100mM potassium tartrate, 50 ⁇ MEGTA, 100 ⁇ MEDTA, 20mM MgCl 2 , 1.7mM ascorbic acid, 2mM ATP, pH7.4. Add ascorbic acid and ATP before the start of the measurement.
  • Washing buffer 25mM HEPES, 100mM potassium tartrate, 50 ⁇ M EGTA, 100 ⁇ M EDTA, pH 7.4.
  • TopSeal-A sealing film Perkin Elmer (Cat#6050185);
  • test results show that, compared with DHTBZ, DHTBZ-X and VBZ, the compound provided by the present invention has stronger in vitro activity.
  • Test Example 2 SD rat pharmacokinetic evaluation of compounds 11, 16, 21, 22, 23 and control examples 44, 45
  • DHTBZ Jiangsu Weikeer Pharmaceutical Technology Co., Ltd., batch number 67-25-1521-59C
  • Test animal SD rat, clean grade, male, weighing about 220g
  • Compound 11, compound 16, compound of control example 44 and DHTBZ were respectively administered to male SD rats (4 animals in each group) by gavage, at a dose of 10 ⁇ mol/kg, 0.25, 0.5, 1, 2, 3 after administration. Approximately 0.2 mL of blood was collected at 4, 6, 8, and 12 hours into a heparinized test tube, and the sample was centrifuged to separate the plasma into a 1.5 ml EP tube within half an hour, and stored at -20°C for testing.
  • VBZ, compound 21, compound 22, compound 23 and the compound of control example 45 were administered to male SD rats (3 animals in each group) by gavage at a dose of 12 ⁇ mol/kg. Blood was collected 0.5h, 2h, and 6h after administration. Put 1mL into a heparinized test tube, centrifuge the sample to separate the plasma into a 1.5ml EP tube within half an hour, and store it at -20°C for testing. After the rats were sacrificed, the brain tissue was taken, and the brain tissue was washed with normal saline, the filter paper was blotted dry, the cerebral blood vessels were peeled off, and the brain tissue was weighed and stored at -20°C.
  • the sample pretreatment adopts the method of protein precipitation, which is briefly described as follows: 25 ⁇ l plasma/50 ⁇ l brain tissue homogenate is used to precipitate protein with 200 ⁇ l/400 ⁇ l acetonitrile containing internal standard, and the supernatant is centrifuged at high speed with water 1: 1(V/V) Dilution and injection analysis.
  • test results of compound 11, compound 16, control 44 compound and DHTBZ administered respectively by intragastric administration to SD rats are shown in Fig. 9.
  • the test results show that compound 11 has a higher exposure than control 44 compound and DHTBZ.
  • test results showed that the brain tissue concentrations of compound 21 and its metabolite compound 11 were significantly higher than VBZ and DHTBZ 2h after administration.
  • DHTBZ Jiangsu Weikeer Pharmaceutical Technology Co., Ltd., batch number 67-25-1521-59C
  • Test animals SD rats, clean grade, male, weighing about 220g, 24 animals, randomly grouped, 3 animals in each group (3 rats by intragastric administration, 3 rats by intravenous administration).
  • Drug solution configuration 21-P3, 11-P4, VBZ and DHTBZ are each accurately weighed at approximately 20 mg, dissolved in an appropriate amount of solvent, and the concentration is 1 ⁇ mol/mL.
  • the LC-MS/MS method was used to determine the concentrations of compounds VBZ, DHTBZ, 21-P3, and 11-P4 in the plasma of SD rats.
  • the concentrations of the corresponding metabolites DHTBZ and 11-P4 in the VBZ and 21-P3 administration groups were simultaneously determined.
  • the sample pretreatment uses the following protein precipitation method: 25 microliters of plasma is precipitated with 200 microliters of acetonitrile containing internal standard, and the supernatant is diluted 1:1 (V/V) with water after high-speed centrifugation and then injected for analysis.
  • Table 31 The average blood concentration (nmol/L) of SD rats after intragastric administration of different compounds (5 ⁇ mol/kg)
  • BLQ means that no samples were taken at this point in time; NA means that data is not available at this point in time.
  • Table 32 The average blood concentration (nmol/L) of SD rats after intravenous administration of different compounds (2 ⁇ mol/kg)
  • BLQ means that no samples were taken at this point in time; NA means that data is not available at this point in time.
  • compound 11-P4 By intravenous administration, compound 11-P4 has a longer half-life, so it can reduce the frequency of administration;
  • the conversion rate of VBZ into DHTBZ was 15.9%, and the conversion rate of compound 21-P3 into 11-P4 was 26.8%. Compared with VBZ, the conversion rate of compound 21-P3 was higher. Since the binding activity of VBZ and 21-P3 to VMAT2 is much lower than that of DHTBZ and 11-P4, the test results show that at an equimolar dose, compared with VBZ, compound 21-P3 will have a higher utilization rate and can produce Higher efficacy.
  • mice SD rats, SPF grade, 32 males, 5-7 weeks old, weight: 200-220g, pre-adapted for at least 1 week before the start of the experiment.
  • Animal source Jinan Pengyue Experimental Animal Technology Co., Ltd.; Animal certificate number: SCXK ( ⁇ )20140007
  • Preparation method Weigh an appropriate amount of the drug, add a small amount of DMSO (not more than 1% of the total volume) to dissolve, add 20% Solutol to the required drug concentration, the final concentration of DMSO is ⁇ 4%
  • mice The day before the test, the rats were randomly divided into 4 groups according to their body weight: control group (NS, no test drug was added to the solvent), VBZ xylene sulfonate group, 21-P3 group, with 8 animals in each group. The rats were pre-adapted to the detection box for 10 minutes and fasted. See Table 3 for animal grouping and dosing information.
  • Rats in each group were given a single intragastric administration of solvent or corresponding drugs according to the dosage in Table 3 and put into the activity room.
  • the TopScan monitoring system was used to record and analyze the total movement distance of the rats within 2hr to 3hr after administration. Avoid putting the same group of rats in the eight activity rooms, and at least one control group rat in each round of the test to prevent mutual interference.
  • the feces were cleaned to avoid the influence of irrelevant interfering factors (odor, etc.) on the movement of rats.
  • test data are expressed as mean ⁇ standard error (MEAN ⁇ SEM), using PASW stastics 18.0 software one-way analysis of variance ANOVA to compare the differences between the groups at each time point, all tests are two-sided tests, P ⁇ 0.05 indicates the difference It is statistically significant.
  • the total exercise reduction rate of VBZ group, 21 group and 21-P3 group were 69.9%, 28.3% and 81.8%, respectively.
  • the 21-P3 group has stronger efficacy.
  • test method is the same as that of Test Example 4. See Table 36 for animal grouping and administration information.
  • 100 ⁇ L incubation system includes: 0.5mg/mL liver microsomes (human, rat, mouse, monkey or dog), 100mM sodium phosphate buffer, 10mM magnesium chloride. Add compounds 11-13, 15-16, and control compound 44 to the incubation system to a final concentration of 1 ⁇ M. After pre-incubating at 37°C for 3min, add NADPH to make the final concentration 1mM, start the reaction, and incubate 0 , 5min, 15min, 30min, 60min, add 200 ⁇ L ice acetonitrile to stop the reaction, store at -20°C for testing. Each sample was processed in parallel at each time point, and a blank control group and a positive control group without coenzyme were set.
  • Test Example 7 Compatibility test of compound 11-P4 and compound 11-P4S crystal form A excipient
  • Compound 11-P4 and compound 11-P4S crystal form A were mixed with excipients in a ratio of 1:20, and then placed in an open place at 60°C for 5 days, and sampled to detect compound 11-P4/compound 11-P4S crystal form The purity of A.
  • Purity detection method take a sample of about 35mg, accurately weigh it, put it in a 25ml measuring flask, add 15ml of acetonitrile to ultrasonically dissolve, dilute to the mark with water, shake it, and use it as the test solution.
  • Test Example 8 11-P4S crystal form A/D/E suspension competition test
  • 11-P4S crystal form A of IPA and IPAc saturated solution respectively add 11-P4S crystal form A/D/E about 5mg each, and suspend and stir at room temperature/50°C for 17 hours to test the solid XRPD.

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Abstract

本发明涉及一类作为VMAT2抑制剂的化合物，具体涉及式(I)所述的化合物或其立体异构体或其药学上可接受的盐及其制备方法，以及在制备治疗与VMAT2相关疾病的药物的应用。(I)

Description

VMAT2抑制剂及其制备方法和应用 技术领域
本发明涉及一类作为VMAT2抑制剂的化合物或其立体异构体或其药学上可接受的盐，及其在与VMAT2相关疾病领域的应用。
背景技术
迟发性运动障碍(tardive dyskinesia，TD)又称迟发性多动症，是1964年由Faurbye首先提出的一种疾病。该病多见于大剂量长期服用抗精神病药物的患者，临床主要特点为不自主、有节律的重复刻板式运动，常累及下颌、嘴唇及舌等；其他药物也可引起迟发性运动障碍，例如帕金森药物(左旋多巴)等。
囊泡单胺转运体2(VMAT2)是位于突触前膜内囊泡膜上的转运蛋白，其功能是将胞浆中单胺递质如多巴胺(DA)或5-羟色胺再摄取进入囊泡内，避免单胺类递质在胞浆内被代谢。VMAT2抑制剂可以拮抗VMAT2的再摄取功能，使多巴胺不能被VMAT2再摄取进囊泡，而被胞浆内的酶代谢掉，致突触间隙的多巴胺释放减少，进而实现治疗迟发性运动障碍的目的。
丁苯那嗪(TBZ，别名丁苯喹嗪)是第一个上市的VMAT2选择性抑制剂，体内的代谢产物反式(2,3)-二氢丁苯那嗪(DHTBZ)同样具备VMAT2选择性抑制活性。2017年4月，FDA批准缬苯那嗪(valbenazine，即VBZ)用于成人迟发性运动障碍的治疗，缬苯那嗪由丁苯那嗪的代谢物DHTBZ酯化而成，较丁苯那嗪的半衰期长，无需频繁给药，其疗效明确，安全性和耐受性良好。
Figure PCTCN2020108314-appb-000001
丁苯那嗪存在半衰期短，需多次给药的问题，并且丁苯那嗪代谢产物种类多，引起的不良反应严重，有抑郁和自杀倾向的黑框不良反应警告。迟发性运动障碍的发病人群较多，但上市药物较少，因此，该领域仍然需要活性更好的VMAT2抑制剂，以满足广泛的临床需求。
发明内容
本发明基于现有技术中存在的问题，提供了式(I)所示化合物或其立体异构体或其药学上可接受的盐。
Figure PCTCN2020108314-appb-000002
其中，
“---”选自：单键或双键；
“---”为单键时，R选自OH、H或
Figure PCTCN2020108314-appb-000003
“---”为双键时，R为O；
R 1选自氢、甲基或乙基；
R 2选自未被取代或者被1、2或3个R 3取代的C 2-10烷基、C 3-6环烷基、C 3-6环烷基-C 1-6烷基、3～6元杂环烷基-C 1-3烷基、C 2-6烯基或C 1-6杂烷基；
R 3选自F、Cl、Br、OH、SH或NH 2。
本发明的一些方案中，式(I)化合物的R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选未被取代或被2-3个R 3取代的C 2-5烷基；R 3为F，其他变量如本发明所定义。
本发明的一些方案中，式(I)化合物的R 2选自乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；优选为三氟乙基或环丙烷亚甲基，其他变量如本发明所定义。
本发明的一些方案中，式(I)化合物的R 1为甲基；“---”为单键，R选自OH或
Figure PCTCN2020108314-appb-000004
或“---”为双键，R为O；其他变量如本发明所定义。
本发明的一些方案中，式(I)化合物所述3～6元杂环烷基或C 1-6杂烷基中的杂原子为O、S或N；杂原子的数目为1-6个，优选为1个O；所述C 1-6杂烷基的C原子数为2-6或3-6或2-5或3-5或4-6或4-5个。
本发明还提供了式(II)所示结构的化合物或其立体异构体或其药学上可接受的盐。
Figure PCTCN2020108314-appb-000005
R 2选自未被取代或者被1、2或3个R 3取代的C 2-10烷基、C 3-6环烷基、C 3-6环烷基-C 1-3烷基、C 2-6烯基、C 1-6杂烷基；R 3选自F、Cl、Br、OH、NH 2。
本发明的一些方案中，式(II)化合物的R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选为未被取代或被2-3个R 3取代的C 2-5烷基；R 3为F。
本发明的一些方案中，式(II)化合物的R 2选自乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；优选为三氟乙基或环丙烷亚甲基。
本发明的一些方案中，式(II)化合物所述C 1-6杂烷基中的杂原子为O、S或N；杂原子的数目为1-6个，优选为1个O。
本发明还提供了式(III)所示结构的化合物或其立体异构体或其药学上可接受的盐，
Figure PCTCN2020108314-appb-000006
R 2选自未被取代或者被1、2或3个R 3取代的C 2-10烷基、C 3-6环烷基、C 3-6环烷基-C 1-3烷基、C 2-6烯基、C 1-6杂烷基；R 3选自F、Cl、Br、OH、NH 2。
本发明的一些方案中，式(III)化合物的R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选为未被取代或被2-3个R 3取代的C 2-5烷基；R 3为F。
本发明的一些方案中，式(III)化合物的R 2选自乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；优选为三氟乙基或环丙烷亚甲基。
本发明的一些方案中，式(III)化合物所述C 1-6杂烷基中的杂原子为O、S或N；杂原子的数目为1-6个，优选为1个O。
本发明提供了式(IV)所示结构的化合物或其立体异构体或其药学上可接受的盐，
Figure PCTCN2020108314-appb-000007
R 2选自未被取代或者被1、2或3个R 3取代的C 2-10烷基、C 3-6环烷基、C 3-6环烷基-C 1-3烷基、C 2-6烯基、C 1-6杂烷基；R 3选自F、Cl、Br、OH、NH 2。
本发明的一些方案中，式(IV)化合物的R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选为未被取代或被2-3个R 3取代的C 2-5 烷基；R 3为F。
本发明的一些方案中，式(IV)化合物的R 2选自乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；优选为三氟乙基或环丙烷亚甲基。
本发明的一些方案中，式(IV)化合物所述C 1-6杂烷基中的杂原子为O、S或N；杂原子的数目为1-6个，优选为1个O。
本发明的一些方案中，还提供了如下结构的化合物或其药学或其对映异构体、非对映异构体、混合物以及药学上可接受的盐：
Figure PCTCN2020108314-appb-000008
Figure PCTCN2020108314-appb-000009
本发明的一些方案中，还提供了如下结构化合物的缬氨酸酯及药学上可接受的盐：
Figure PCTCN2020108314-appb-000010
本发明的一些方案中，还提供了如下结构的化合物或其药学上可接受的盐：
Figure PCTCN2020108314-appb-000011
本发明还提供了上述任意一种化合物的对甲基苯磺酸盐，优选以下化合物：
Figure PCTCN2020108314-appb-000012
本发明还提供了化合物11-P4S五种晶型。
本发明的一些方案中，11-P4S晶型A属于正交晶系P21212空间群，晶胞参数
Figure PCTCN2020108314-appb-000013
Figure PCTCN2020108314-appb-000014
α＝90°,β＝90°,γ＝90°,
Figure PCTCN2020108314-appb-000015
Z＝4。
本发明的一些方案中，11-P4S晶型A通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.33±0.2°，10.87±0.2°，18.89±0.2°。
本发明的一些方案中，11-P4S晶型A通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.33±0.2°，10.87±0.2°，16.61±0.2°，18.89±0.2°，19.27±0.2°，22.19±0.2°。
本发明的一些方案中，11-P4S晶型A通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.33±0.2°，10.87±0.2°，13.77±0.2°，16.61±0.2°，18.20±0.2°，18.89±0.2°，19.27±0.2°，20.05±0.2°，22.19±0.2°，24.60±0.2°，24.77±0.2°。
本发明的一些方案中，11-P4S晶型A，通过Cu-Ka辐射具有基本如图2-1所示的X射线粉末衍射图谱。
本发明的一些方案中，11-P4S晶型B通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.32±0.2°，5.42±0.2°，10.85±0.2°。
本发明的一些方案中，11-P4S晶型B通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.32±0.2°，5.42±0.2°，10.85±0.2°，16.60±0.2°，18.88±0.2°，22.02±0.2°。
本发明的一些方案中，11-P4S晶型B，通过Cu-Ka辐射具有基本如图3-1所示的X射线粉末衍射图谱。
本发明的一些方案中，11-P4S晶型C通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：5.81±0.2°，6.33±0.2°，12.86±0.2°。
本发明的一些方案中，11-P4S晶型C通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：5.81±0.2°，6.33±0.2°，7.99±0.2°，12.86±0.2°，19.09±0.2°，23.17±0.2°。
本发明的一些方案中，11-P4S晶型C通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：5.81±0.2°，6.33±0.2°，7.99±0.2°，10.31±0.2°，11.63±0.2°，12.86±0.2°，18.16±0.2°，19.09±0.2°，23.17±0.2°，24.00±0.2°，27.32±0.2°。
本发明的一些方案中，11-P4S晶型C，通过Cu-Ka辐射具有基本如图4-1所示的X射线粉末衍射图谱。
本发明的一些方案中，11-P4S晶型D通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.02±0.2°，23.91±0.2°。
本发明的一些方案中，11-P4S晶型D通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：5.31±0.2°，6.02±0.2°，18.88±0.2°，22.12±0.2°，23.91±0.2°。
本发明的一些方案中，11-P4S晶型D，通过Cu-Ka辐射具有基本如图5-1所示的X射线粉末衍射图谱。
本发明的一些方案中，11-P4S晶型E通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.06±0.2°，18.32±0.2°，30.79±0.2°
本发明的一些方案中，11-P4S晶型E，通过Cu-Ka辐射具有基本如图6-1所示的X射线粉末衍射图谱。
本发明还提供了19P2对甲基苯磺酸盐(简称为19P2S)的晶型，该晶型属于正交晶系P212121空间群。晶胞参数
Figure PCTCN2020108314-appb-000016
α＝90°,β＝90°,γ＝90°,
Figure PCTCN2020108314-appb-000017
Z＝4。
本发明还提供了一种药物组合物，包括上述任一所述化合物或其立体异构体或其药学上可接受的盐，或上述任一所述化合物的结晶形式以及药学上可接受的载体。所述药物组合物可以制备成药学上可接受的各种剂型，如片剂、胶囊剂、口服液剂、颗粒剂、注射剂或各种缓控释制剂等。所述药物组合物可通过口服或胃肠外方式(如静脉内、皮下或局部等)给药。给药剂量可根据患者的年龄、性别和疾病类型进行适当调整，一般每日剂量约为10-100mg/天。
本发明还提供了上述任一所述化合物或其立体异构体或其药学上可接受的盐，或上述任一所述化合物的结晶形式，或药物组合物在制备治疗与VMAT2相关疾病的药物的应用。
本发明还提供了上述化合物或其立体异构体或其药学上可接受的盐，或上述任一所述化 合物的结晶形式，或药物组合物在制备治疗运动机能亢进性障碍的药物上的应用；优选所述运动机能亢进性障碍包括亨廷顿舞蹈症、迟发性运动障碍、图雷特综合征或抽搐。
本发明还提供了式(II)化合物的制备方法，包含以下制备步骤，
路线1：
Figure PCTCN2020108314-appb-000018
路线2：
Figure PCTCN2020108314-appb-000019
路线3：
Figure PCTCN2020108314-appb-000020
其中X为离去基团，R 2与上述式(II)化合物中的定义相同，优选R 2为乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；更优选为三氟乙基或环丙烷亚甲基。
本发明还提供了式(III)化合物的制备方法，包含下述步骤，
Figure PCTCN2020108314-appb-000021
其中以硼氢化钠作为还原试剂，R 2与上述式(III)化合物中的定义相同，优选R 2为乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；更优选为三氟乙基或环丙烷亚甲基。
本发明还提供了式(IV)化合物的制备方法，包含下述步骤，
Figure PCTCN2020108314-appb-000022
其中，R 2与上述式(IV)化合物中的定义相同，优选R 2为乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；更优选为三氟乙基或环丙烷亚甲基。P基团为氨基保护基，优选为苄氧羰基(Cbz)，叔丁氧基羰基(Boc)，芴甲氧羰基(Fmoc)，烯丙氧羰基(Alloc)，三甲基硅乙氧羰基(Teoc)等，脱出保护基可采用本领域常用的方法，具体可参见Protective Groups in Organic Synthesis,Third Edition，Author(s):Theodora W.Greene Ph.D.，第7章。
本发明还提供了式(I)化合物立体异构体的的制备方法，具体如下：
Figure PCTCN2020108314-appb-000023
其中，所述R为-OH或
Figure PCTCN2020108314-appb-000024
其他变量与上述式(I)化合物中的定义相同；所述制备方法为使用手性色谱柱对式(I)化合物进行拆分，手性色谱柱优选为大赛璐CHIRALPAKAD-H色谱柱。
本发明提供的化合物具有以下任意一种或多种优势：与VMAT2亲和力强，体内具有更高的暴露量，脑内浓度高、半衰期长，药效强等。
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。当本文中出现商品名时，意在指代其对应的商品或其活性成分。
这里所采用的术语“药学上可接受的”，是针对化合物、材料、组合物和/或剂型而言，它们在可靠的医学判断的范围之内，适用于与人类和动物的组织接触使用，而没有过多的毒性、刺激性、过敏性反应或其它问题或并发症，与合理的利益/风险比相称。
术语“药学上可接受的盐”是指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物的中性形式接触的方式获得碱加成盐。药学上可接受的碱加成盐包括钠、钾、钙、铵、有机氨或镁盐或类似的盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸 与这类化合物的中性形式接触的方式获得酸加成盐。药学上可接受的酸加成盐的实例包括无机酸盐，或有机酸盐。本发明的某些特定的化合物含有碱性和酸性的官能团，从而可以被转换成任一碱或酸加成盐。
本发明的药学上可接受的盐可由含有酸根或碱基的母体化合物通过常规化学方法合成。一般情况下，这样的盐的制备方法是：在水或有机溶剂或两者的混合物中，经由游离酸或碱形式的这些化合物与化学计量的适当的碱或酸反应来制备。
本发明的化合物可以存在特定的几何或立体异构体形式。本发明设想所有的这类化合物，包括顺式和反式异构体、(-)-和(+)-对对映体、(R)-和(S)-对映体、非对映异构体、(D)-异构体、(L)-异构体，及其外消旋混合物和其他混合物，例如对映异构体或非对映体富集的混合物，所有这些混合物都属于本发明的范围之内。烷基等取代基中可存在另外的不对称碳原子。所有这些异构体以及它们的混合物，均包括在本发明的范围之内。
除非另有说明，术语“对映异构体”是指互为镜像关系的立体异构体。
除非另有说明，术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。
除非另有说明，“(D)”或者“(+)”表示右旋，“(L)”或者“(-)”表示左旋，“(DL)”或者“(±)”表示外消旋。
除非另有说明，用楔形实线键
Figure PCTCN2020108314-appb-000025
和楔形虚线键
Figure PCTCN2020108314-appb-000026
表示一个立体中心的绝对构型。
可以通过的手性合成或手性试剂或者其他常规技术制备光学活性的(R)-和(S)-异构体以及D和L异构体。如果想得到本发明某化合物的一种对映体，可以通过不对称合成或者具有手性助剂的衍生作用来制备，其中将所得非对映体混合物分离，并且辅助基团裂开以提供纯的所需对映异构体。或者，当分子中含有碱性官能团(如氨基)或酸性官能团(如羧基)时，与适当的光学活性的酸或碱形成非对映异构体的盐，然后通过本领域所公知的常规方法进行非对映异构体拆分，然后回收得到纯的对映体。此外，对映异构体和非对映异构体的分离通常是通过使用色谱法完成的，所述色谱法采用手性固定相，并任选地与化学衍生法相结合(例如由胺生成氨基甲酸盐)。本发明的化合物可以在一个或多个构成该化合物的原子上包含非天然比例的原子同位素。例如，可用放射性同位素标记化合物，比如氚(3H)，碘-125(125I)或C-14(14C)。又例如，可用重氢取代氢形成氘代药物，氘与碳构成的键比普通氢与碳构成的键更坚固，相比于未氘化药物，氘代药物有降低毒副作用、增加药物稳定性、增强疗效、延长药物生物半衰期等优势。本发明的化合物的所有同位素组成的变换，无论放射性与否，都包括在本发明的范围之内。
附图说明
图1：化合物11-P4S分子结构椭球图
图2-1、2-2、2-3：分别为化合物11-P4S晶型A的XRPD图、TGA/DSC图、 1H NMR图
图3-1、3-2、3-3：分别为化合物11-P4S晶型B的XRPD图、TGA/DSC图、 1H NMR图
图4-1、4-2、4-3：分别为化合物11-P4S晶型C的XRPD图、TGA/DSC图、 1H NMR图
图5-1、5-2、5-3：分别为化合物11-P4S晶型D的XRPD图、TGA/DSC图、 1H NMR图
图6-1、6-2、6-3：分别为化合物11-P4S晶型E的XRPD图、TGA/DSC图、 1H NMR图
图7：化合物11-P3S分子结构椭球图
图8：化合物19P2S分子结构椭球图
图9：化合物11、化合物16、对照例44化合物和DHTBZ分别灌胃给药雄性SD大鼠后的血药浓度-时间曲线图
图10：VBZ、化合物21、化合物22、化合物23和对照例45化合物灌胃给药SD大鼠后，原化合物脑组织中药物分布
图11：活性代谢物(DHTBZ、化合物11、化合物12、化合物13和对照例44化合物)脑组织中药物组织分布
图12：VBZ、化合物21、化合物22、化合物23和对照例45化合物灌胃给药SD大鼠后，原化合物脑血比(B/P ratio)；
图13：活性代谢物(DHTBZ、化合物11、化合物12、化合物13和对照例44化合物)脑血比(B/P ratio)
图14：VBZ及11-P4的剂量效应曲线
图15：11-P4S晶型A/D/E混悬竞争结果的XRPD叠图
具体实施方式
实施例1
片段1的制备：
Figure PCTCN2020108314-appb-000027
合成路线：
Figure PCTCN2020108314-appb-000028
1、将化合物a(5.0g，32.9mmol)溶于50mL DMF中，加入溴化苄(6.2g，35.8mmol)，碳酸钾(6.8g，49.1mmol)，室温搅拌反应过夜。向反应体系中加入100mL水，有固体析出，抽滤得7.2g白色固体b。
2、将化合物b(5.4g，22.3mmol)溶于50mL硝基甲烷中，加入乙酸铵(0.86g，11.2mmol)，加热至110℃搅拌反应3h。反应体系冷却至室温，加入100mL水，有固体析出，抽滤得6.0g黄色固体c。
3、在氮气保护下，将氢化铝锂(2.0g，52.6mmol)缓慢加入到50mL无水四氢呋喃中，降温至0℃，慢慢滴加c(5.0g，17.5mmol)，然后将反应体系升温至60℃搅拌反应2h。冷却至0℃，用水淬灭反应，抽滤，将滤液用无水硫酸钠干燥，然后减压浓缩得3.2g淡黄色油状液体d。产品不经纯化直接进行下一步反应。MS m/z(ESI)：258.2[M+1]
4、将粗品d(2.7g，10.5mmol)溶解于16mL冰醋酸中，加入4mL三氟乙酸，然后加入乌洛托品(3.0g，21.0mmol)，升温至80℃搅拌反应2h。冷却至室温，加入50g碎冰，然后用20％氢氧化钠溶液调节PH至8。用二氯甲烷萃取萃取(50mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，得到2.4g粗品产物e。产品不经纯化直接进行下一步反应。MS m/z(ESI)：268.1[M+1]
5、将粗品e(2.0g，5.1mmol)溶解于20mL乙醇和20mL水的体系中，加入苄基三乙基氯化铵(0.43g，1.3mmol)，加热至回流搅拌反应5h。将溶剂减压蒸出，残余物加入50mL水，然后用乙酸乙酯萃取(20mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩得粗产物。然后用乙醇进行重结晶，得到0.8g白色固体化合物f。MS m/z(ESI)：394.2[M+H] +；1H NMR(600MHz,CDCl3):δ7.43-7.28(m,5H),6.64(s,1H),6.57(s,1H),5.11(s,2H),3.82(s,3H),3.48(dd,1H),3.26(dd,1H),3.13-3.00(m,2H),2.90(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.33(t,1H),1.83-1.75(m,1H),1.70-1.58(m,1H),1.06-0.98(m,1H),0.90(m,6H)。
6、将化合物f(0.5g，1.3mmol)溶于20mL甲醇中，加入0.05g钯碳，在氢气氛围下，室温搅拌8h。过滤除去钯碳，将滤液减压蒸出溶剂得0.47g淡黄色粉末片段1。MS m/z(ESI)：304.2[M+1]。
化合物1的制备：
Figure PCTCN2020108314-appb-000029
合成路线：
Figure PCTCN2020108314-appb-000030
将片段1化合物(150mg，0.50mmol)溶于2mLDMF中，加入3-溴丙基甲基醚(84mg，0.55mmol)、碳酸钾(103mg，0.75mmol)，升温至60℃搅拌反应5h。冷却后，向反应体系中 加入8mL水，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝3:1)，得化合物1(120mg，淡黄色蜡状固体)。MS m/z(ESI)：376.2[M+H] +； 1H NMR(600MHz，CDCl 3):δ6.65(s,1H),6.54(s,1H),4.07(t,2H),3.80(s,3H),3.55(t,2H),3.48(dd,1H),3.34(s,3H),3.26(dd,1H),3.15-3.02(m,2H),2.88(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.33(t,1H),2.11-2.04(m,2H),1.83-1.75(m,1H),1.70-1.58(m,1H),1.06-0.98(m,1H),0.90(m,6H).
实施例2：
Figure PCTCN2020108314-appb-000031
合成路线：
Figure PCTCN2020108314-appb-000032
将片段1化合物(150mg，0.50mmol)溶于2mL DMF中，加入1-溴-4-氟丁烷(85mg，0.55mmol)、碳酸钾(103mg，0.75mmol)，升温至60℃搅拌反应5h。冷却后，向反应体系中加入8mL水，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝3:1)，得化合物2(115mg，淡黄色油状液体)。MS m/z(ESI)：378.2[M+H] +； 1H NMR(600MHz，CDCl 3):δ6.64(s,1H),6.54(s,1H),4.57(t,1H),4.45(t,1H),4.07(t,2H),3.80(s,3H),3.48(dd,1H),3.26(dd,1H),3.15-3.02(m,2H),2.88(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.33(t,1H),1.96-1.90(m,3H),1.88-1.75(m,2H),1.70-1.58(m,1H),1.06-0.98(m,1H),0.89(m,6H).
实施例3：
Figure PCTCN2020108314-appb-000033
合成路线：
Figure PCTCN2020108314-appb-000034
将片段1化合物(150mg，0.50mmol)溶于2mL DMF中，加入溴甲基环丙烷(74mg，0.55mmol)、碳酸钾(103mg，0.75mmol)，升温至60℃搅拌反应5h。冷却后，向反应体系中加入8mL水，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝3:1)，得化合物3(107mg，淡黄色蜡状固体)。MS m/z(ESI):358.2[M+H] +； 1H NMR(600MHz，CDCl 3):δ6.61(s,1H),6.54(s,1H),3.82-3.79(m,5H),3.48(dd,1H),3.28(dd,1H),3.15-3.02(m,2H),2.89(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.33(t,1H),1.82-1.75(m,1H),1.70-1.58(m,1H),1.06-0.98(m,1H),0.89(m,6H),0.65-0.59(m,2H),0.35-0.30(m,2H).
实施例4：
Figure PCTCN2020108314-appb-000035
合成路线：
Figure PCTCN2020108314-appb-000036
将片段1化合物溶于2mL DMF中，加入溴丙烷(68mg，0.55mmol)、碳酸钾(103mg，0.75mmol)，升温至60℃搅拌反应5h。冷却后，向反应体系中加入8mL水，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝3:1)，得化合物4(102mg，淡黄色蜡状固体)。MS m/z(ESI)：346.2[M+H] +； 1H NMR(600MHz,CDCl 3):δ6.64(s,1H),6.54(s,1H),33.96-3.90(t,2H),3.81(s,3H),3.49(dd,1H),3.27(dd,1H),3.15-3.02(m,2H),2.88(dd,1H),2.74-2.66(m,2H),2.62-2.48(m,2H),2.33(t,1H),1.96-1.90(m,3H),1.70-1.58(m,1H),1.06-0.98(m,4H),0.89(m,6H).
实施例5：
Figure PCTCN2020108314-appb-000037
合成路线：
Figure PCTCN2020108314-appb-000038
20mL微波管中，将片段1化合物(606mg，2.0mmol)溶于6mL DMF中，加入1,1,-二氟-2-碘乙烷(768mg，4.0mmol)、碳酸钾(1.10g，8.0mmol)，升温至100℃，反应3h。冷却后，将反应体系倾入30mL水中，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得化合物5(600mg，类白色固体)。MS m/z(ESI)：368.2[M+H] +； 1H NMR(400MHz,CDCl 3):δ6.68(s,1H),6.59(s,1H),5.97-6.24(m,1H),4.16-4.23(m,2H),3.82(s,3H),3.42-3.52(m,1H),3.16-3.31(m,1H),2.54-3.12(m,7H),2.32-2.38(m,1H),1.63-1.71(m,1H),1.00-1.07(m,1H),0.88-0.92(m,6H).
实施例6：
Figure PCTCN2020108314-appb-000039
合成路线1：
Figure PCTCN2020108314-appb-000040
20mL微波管中，将片段1化合物(1.82g，6.0mmol)溶于15mL DMF中，加入1,1,1-三氟-2-碘乙烷(2.30g，12mmol)、碳酸钾(3.31g，24mmol)，升温至140℃，反应3h。冷却后，将反应体系倾入75mL水中，然后用乙酸乙酯萃取(15mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得化合物6(610mg，类白色固体)。MS m/z(ESI)：386.2[M+H] +； 1H NMR(400MHz,CDCl 3)δ6.75(s,1H),6.60(s,1H),4.35-4.39(m,2H),3.82(s,3H),3.49-3.53(m,1H),3.26-3.31(m,1H),2.38-3.15(m,7H), 2.32-2.35(m,1H),1.37-1.84(m,2H),1.00-1.08(m,1H),0.90-0.92(m,6H).
合成路线2：
Figure PCTCN2020108314-appb-000041
步骤1：将反应物I(50mg，0.28mmol)溶于DMF(1mL)，加入K 2CO 3(77mg，0.56mmol)，室温搅拌反应0.5h。加入三氟碘乙烷(76mg,0.36mmol)，80℃搅拌反应过夜，TLC检测反应完全。加水淬灭反应，析出固体，搅拌1h，抽滤，滤饼用水洗两次。收集滤饼，干燥，得中间体1，微黄色固体0.07g。
步骤2：将中间体1(0.07g，0.270mmol)溶于1mL乙醇和1mL水混合溶液中，加入3-二甲基氨基-5-甲基-2-己酮(0.06g，0.324mmol)，苄基三乙基氯化铵(0.02g，0.081mmol)加热至95℃反应18h。冷却至室温浓缩，残余物用乙酸乙酯(20mL*3)萃取，合并有机相，有机相无水硫酸钠干燥，过滤，减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝5:1)，得化合物6(10mg，类白色固体)，产率：10％。
合成路线3：
Figure PCTCN2020108314-appb-000042
步骤1：将反应物I(50mg，0.282mmol)溶于1mL乙醇和1mL水混合溶液中，加入3-二甲基氨基-5-甲基-2-己酮(0.06g，0.338mmol)，苄基三乙基氯化铵(0.02g，0.085mmol)加热至95℃反应18h。冷却至室温浓缩，残余物用乙酸乙酯(20mL*3)萃取，合并有机相，有机相无水硫酸钠干燥，过滤，减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝5:1)，得片段1(70mg，类白色固体)，产率：82％。
步骤2：与合成路线1相同。
实施例7：
Figure PCTCN2020108314-appb-000043
合成路线：
Figure PCTCN2020108314-appb-000044
将片段1化合物(606mg，2.0mmol)溶于6mL DMF中，加入1-溴-4，4,4-三氟丁烷(573mg，3.0mmol)、碳酸钾(828mg，6.0mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入30mL水中，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得化合物7(480mg，类白色固体)。MS m/z(ESI)：414.2[M+H] +； 1H NMR(400MHz,CDCl 3)δ6.62(s,1H),6.56(s,1H),4.03-4.05(m,2H),3.81(s,3H),3.49-3.51(m,1H),3.21-3.42(m,1H),2.54-3.14(m,7H),2.29-2.37(m,3H),2.05-2.09(m,2H),1.68-1.83(m,1H),1.64-1.68(m,1H),1.01-1.06(m,1H),0.90-0.92(m,6H).
实施例8：
Figure PCTCN2020108314-appb-000045
合成路线：
Figure PCTCN2020108314-appb-000046
将片段1化合物(100mg，0.33mmol)溶于1mL DMF中，加入1 1-溴-2-氟乙烷(62.7mg，0.5mmol)、碳酸钾(138mg，1mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入5mL水中，然后用乙酸乙酯萃取(2mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(二氯甲烷：甲醇＝80:1)，得化合物8(75mg，类白色固体)。MS m/z(ESI)：350.2[M+H] +； 1H NMR(400MHz,CDCl 3)δ6.68(s,1H),6.57(s,1H),4.72-4.81(m,2H),4.22-4.28(m,2H),3.82(s,3H),2.29-3.58(m,10H),1.64-1.83(m,2H),1.02-1.06(m,1H),0.90-0.93(m,6H).
实施例9：
Figure PCTCN2020108314-appb-000047
合成路线：
Figure PCTCN2020108314-appb-000048
将片段1化合物(606mg，2.0mmol)溶于6mL DMF中，加入溴代异丁烷(411mg，3.0mmol)、碳酸钾(828mg，6.0mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入30mL水中，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得化合物9(410mg，类白色固体)。MS m/z(ESI)：360.2[M+H] +； 1H NMR(400MHz,CDCl 3)δ6.62(s,1H),6.55(s,1H),3.83(s,3H),3.70-3.80(m,2H),2.35-3.50(m,10H),2.11-2.18(m,1H),1.78-1.83(m,1H),1.64-1.69(m,1H),1.02-1.06(m,7H),0.90-0.92(m,6H).
实施例10：
Figure PCTCN2020108314-appb-000049
合成路线：
Figure PCTCN2020108314-appb-000050
将片段1化合物(606mg，2.00mmol)溶于6mL DMF中，加入溴乙烷(327mg，3.00mmol)、碳酸钾(414mg，3.0mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入30mL水中，然后用乙酸乙酯萃取(20mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得化合物10(397mg，类白色固体)。MS m/z(ESI)：332.2[M+H] +；
实施例11：
Figure PCTCN2020108314-appb-000051
合成路线：
Figure PCTCN2020108314-appb-000052
氮气保护，冰浴下，将化合物6(610mg，1.58mmol)溶于10mL THF中，加入硼氢化钠(120mg，3.17mmol)、10mL MeOH，冰浴下反应2h。加入1N盐酸淬灭，加入NaHCO 3水溶液调pH值至碱性，浓缩有机相，然后用乙酸乙酯萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，减压浓缩有机相，用柱层析分离(二氯甲烷：甲醇＝40:1)，得化合物11(346mg，类白色固体)，收率：56.5％。MS m/z(ESI)：388.2[M+H] +； 1H NMR(400MHz,CD 3OD)δ6.87(s,1H),6.79(s,1H),4.43-4.49(m,2H),3.85(s,3H),2.93-3.24(m,4H),2.44-2.71(m,3H),1.02-2.10(m,7H),0.94-1.02(m,6H)。
实施例12-20：
实施例12-20的合成采用和实施例11类似的方法进行。
表1：实施例12-20化合物结构和表征数据
Figure PCTCN2020108314-appb-000053
Figure PCTCN2020108314-appb-000054
实施例21：
Figure PCTCN2020108314-appb-000055
合成路线：
Figure PCTCN2020108314-appb-000056
1、氮气保护，冰浴下，将Boc-L-缬氨酸(857.6mg，3.95mmol)溶于20mL二氯甲烷中，加入4-二甲氨基吡啶(386.9mg，3.16mmol)，化合物11(1.02g，2.64mmol)，冰浴下搅拌反 应5分钟。一次性加入二环己基碳二亚胺(813.7mg，3.95mmol)，自然升温，反应18小时。过滤，滤液减压浓缩，用柱层析分离(二氯甲烷：甲醇＝30:1)纯化所得残余物，得化合物21a(1.13g，类白色固体)。MS m/z(ESI)：587.3[M+H] +
2、将化合物21a(1.13g，1.92mmol)，溶于15ml浓度为4M的1,4-二氧六环溶液中，室温下反应2小时。浓缩反应液，乙醚(10ml*1)洗涤固体，得粗品。将粗品溶于30mL水中，饱和碳酸氢钠水溶液调Ph＝7～8,二氯甲烷(10ml*3)萃取，合并有机相。水(10ml*1)和饱和氯化钠(10ml*1)分别洗涤二氯甲烷相。无水硫酸钠干燥二氯甲烷相。过滤，滤去固体，浓缩二氯甲烷相，得化合物21(720mg)。MS m/z(ESI)：487.3[M+H] +；1H NMR(400MHz,CD 3OD)δ6.76(s,1H),6.74(s,1H),4.65-4.74(m,1H),4.38-4.45(m,2H),3.77(s,3H),3.25-3.27(m,1H),2.97-3.12(m,3H),2.61-2.74(m,2H),2.47-2.51(m,1H),1.94-2.16(m,3H),1.63-1.75(m,1H),1.43-1.51(m,1H),1.27-1.37(m,1H),1.01-1.08(m,2H),0.88-1.00(m,12H)。
实施例22-25：
实施例22-25的合成采用和实施例21类似的方法进行。
表2-1：实施例22-25的化合物结构和表征数据
Figure PCTCN2020108314-appb-000057
Figure PCTCN2020108314-appb-000058
实施例26-28：
化合物26-28的合成采用和实施例11类似的方法进行。
表2-2：实施例22-25的化合物结构和表征数据
Figure PCTCN2020108314-appb-000059
实施例29：
Figure PCTCN2020108314-appb-000060
Figure PCTCN2020108314-appb-000061
步骤1：中间体I的合成：
在2L单口瓶中加入原料片段I(106g,350mmol,1eq)和无水碳酸钾(145g,1050mmol,3eq),加入无水N,N-二甲基甲酰胺(700mL),然后加入1,1,1-三氟-2-碘乙烷(184g,875mmol,2.5eq)，140度下反应11h。TLC(PE:EA＝3:1)监控显示仍有部分原料剩余，补加1,1,1-三氟-2-碘乙烷(73.5g,350mmol,1eq)，140度下继续反应8h。反应完全，冷却至室温，反应体系倾入至倾入水(3.5L)和饱和食盐水(700mL)的混合溶液中，用乙酸乙酯(700ml)萃取四次，合并乙酸乙酯相。乙酸乙酯相分别用水(700mL)以及饱和氯化钠溶液(700mL)洗一次。无水硫酸钠干燥乙酸乙酯相，过滤，滤去无水硫酸钠，减压浓缩乙酸乙酯相。残留物用乙酸乙酯(175mL)和石油醚(175mL)的混合溶液打浆，过滤，收集固体。固体每次用乙酸乙酯(175mL)和石油醚(175mL)的混合溶液洗涤，共洗涤三次。干燥后，得50g淡黄色固体。将50g淡黄色固体溶于回流状态的95％乙醇(700mL)中，自然降温4h，析出晶体。过滤，收集晶体，每次用室温状态的95％乙醇(175mL)洗涤晶体，共洗涤三次。干燥后，得中间体I(38.4g，淡黄色晶体)，收率：28.5％。
MS m/z(ESI)：386.2[M+H] +， 1H NMR(600MHz,CDCl 3)：δ6.75(s,1H),6.59(s,1H),4.38-4.34(m,2H),3.82(s,3H),3.53-3.51(m,1H),3.32-3.28(m,1H),3.15-3.09(m,2H),2.91-2.88(m,1H),2.75-2.72(m,2H),2.61-2.53(m,2H),2.38-2.34(m,1H),1.82-1.78(m,1H),1.69-1.62(m,1H),1.06-1.01(m,1H),0.92-0.90(m,6H).
步骤2：化合物11的合成：
在1L单口瓶中，加入中间体I(38.46g，100mmol,1eq)，四氢呋喃(150mL)，冰浴下，加入硼氢化钠(4.56g,120mmol,1.2eq)，然后加入无水甲醇(150ml)，冰浴下搅拌2h。反应完全，氮气氛围下，冰浴下，用1N HCL(300mL,300mmol,3eq)淬灭反应。冰浴下，滴加碳酸钠的饱和溶液(300mL)，生成淡黄色固体。过滤，收集固体。每次用水(300mL)洗涤固体，共洗涤三次。干燥后，得到39g淡黄色固体。固体用80度的乙酸乙酯(300mL)溶解，然后加入石油醚(100mL)，自然降温4h，析出白色固体。收集固体，每次用乙酸乙酯(50mL)和石油醚(50mL)的混合溶液洗涤固体，共洗涤三次。干燥后，得化合物11(23.07g，白色固体)，收率：59.6％。MS m/z(ESI)：388.2[M+H] +； 1H NMR(600MHz,CDCl 3)：δ6.71(s,1H),6.70(s,1H),4.38-4.31(m,2H),3.82(s,3H),3.41-3.35(m,1H),3.13-2.96(m,4H),2.64-2.54(m,2H),2.47-2.40(m,1H),2.04-1.94(m,1H),1.75-1.66(m,3H),1.61-1.45(m,2H),1.08-1.02(m,1H),0.92-0.90(m,6H).
步骤3：化合物11的拆分
取12.33g化合物11，采用HPLC法，使用大赛璐的制备色谱和大赛璐手性柱对手性异构体分离，收集其相应组分，旋转蒸发除去溶剂，得到光学异构体纯品。分离方法和检测结果 见表3-4。
表3：化合物11的手性分离方法
Figure PCTCN2020108314-appb-000062
表4：化合物11的手性分析结果
峰号 保留时间 峰面积 相对峰面积％
1 4.994 5152535 48.604
2 5.601 114788 1.083
3 6.109 5177576 48.840
4 6.707 156247 1.474
分别收集11-P4(保留时间4.994min)和11-P3(保留时间6.109min)的组分，旋转蒸发除去溶剂，分别得到6.1448g的11-P4和5.7844g的11-P3样品，分析方法和结果见表5-表8。
表5：化合物11-P4的分析方法
色谱柱 CHIRALPAK AD-H(ADH0CD-UE022)
色谱柱尺寸 0.46cm I.D.×15cm L
进样量 0.5ul
流动相 Hexane/EtOH＝60/40(V/V)
流速 1.0ml/min
检测波长 UV 214nm
柱温 35℃
HPLC设备 Shimadzu LC-20AT CP-HPLC-09
样品名称 11-P4
表6：化合物11-P4的分析结果
峰号 保留时间 峰面积 相对峰面积％
1 1.920 502928 3.811
2 4.939 12433977 94.208
3 6.139 29066 0.220
4 6.440 232459 1.761
表7：化合物11-P3的分析方法
色谱柱 CHIRALPAK AD-H(ADH0CD-UE022)
色谱柱尺寸 0.46cm I.D.×15cm L
进样量 1ul
流动相 Hexane/EtOH＝60/40(V/V)
流速 1.0ml/min
检测波长 UV 214nm
柱温 35℃
HPLC设备 Shimadzu LC-20AT CP-HPLC-09
样品名称 11-P3
表8：化合物11-P3的分析结果
峰号 保留时间 峰面积 相对峰面积％
1 1.919 512872 5.985
2 4.999 150632 1.758
3 6.056 7906161 92.258
化合物11-P4与11-P3的质谱、氢谱以及碳谱相同：
MS m/z(ESI)：388.2[M+H] +；
1HNMR(600MHz,CDCl 3)：δ6.72(s,1H),6.71(s,1H),4.33-4.37(q,J＝8.0Hz,2H),3.82(s,3H),3.37-3.40(m,1H),3.12-3.14(d,J＝12.0Hz,1H),3.06-3.08(m,1H),3.02-3.05(m,1H),2.98-3.00(m,1H),2.60-2.63(m,1H),2.55-2.58(m,1H),2.45-2.46(m,1H),1.95-1.99(t,J＝12.0Hz,1H),1.72-1.74(m,1H),1.68-1.71(m,1H),1.55-1.60(m,1H),1.47-1.53(m,1H),1.03-1.07(m,1H),0.91-0.92(d,J＝6.0Hz,3H),0.93-0.94(d,J＝6.0Hz,3H).
13CNMR(150Hz,CDCl 3)：δ148.5,145.53,132.85,126.84,120.80-126.34,117.66,109.29,74.46,67.66-68.36,60.94,59.99,56.09,51.74,41.51,40.44,39.64,28.83,25.33,24.15,21.74.
实施例30
I.化合物11-P4S晶型A
步骤1：化合物11-P4成单对甲苯磺酸盐：
Figure PCTCN2020108314-appb-000063
将11-P4(0.20g，0.52mmol)溶于乙酸乙酯(5ml)中，滴加对甲苯磺酸一水合物(0.12g,0.62mmol)的乙酸乙酯溶液，析出白色固体。室温搅拌12h，抽滤，滤饼用乙酸乙酯洗涤(5mL*3)，滤饼干燥，得化合物11-P4S白色固体0.22g，收率78％。
步骤2：化合物11-P4S晶型A的单晶培养方法
1)称取9mg的11-P4S于1.5mL HPLC小瓶中。
2)向固体中加入450微升的乙醇，升温至40℃，并在40℃下恒温直至固体全部溶清。
3)静置状态下将溶液以0.3℃/min降至25度。
4)析出晶体，将反应瓶拿到显微镜下观察，晶体合格，进行XRSD试验。
步骤3：化合物11-P4S晶型A的XRSD试验：
3.1仪器参数与数据收集：
3.1.1仪器参数：
单晶衍射仪：Rigaku Oxford Diffraction XtaLAB Synergy four-circle diffractometer
探测器：HyPix-6000HE面探测器；
低温系统:Oxford Cryostream 800；
光源：Cu靶微焦斑光源；
Figure PCTCN2020108314-appb-000064
50W；
晶体与CCD探测器距离:d＝35mm；
管压:50kV；
管流:1mA
3.1.2数据收集：
衍射试验收集了48459个衍射点，其中独立衍射点4803个(Rint＝0.0672)；衍射收集范围2θ＝6.342to 133.2°，衍射指标范围-32≤h≤32,-19≤k≤19,-7≤l≤6。结构解析使用SHELXT(Sheldrick,G.M.2015.ActaCryst.A71,3-8)，结构精修使用SHELXL(against F 2)(Sheldrick,G.M.2015.ActaCryst.C71,3-8)。4803个独立衍射点中，参加结构精修的参数为348。精修后S＝1.046,R 1＝0.0318,wR 2＝0.0828。残余电子密度值为0.26和
Figure PCTCN2020108314-appb-000065
3.2数据列表见表9和表10
表9：化合物11-P4S晶型A的单晶衍射数据列表
Figure PCTCN2020108314-appb-000066
Figure PCTCN2020108314-appb-000067
表10：原子坐标(x 10^4)与等效各向同性位移参数((A^2 x 10^3)
Figure PCTCN2020108314-appb-000068
3.3结论
化合物11-P4S晶型A为无色块状(0.20×0.10×0.10mm 3)，属于正交晶系P21212空间群。晶胞参数：
Figure PCTCN2020108314-appb-000069
α＝90°,β＝90°,γ＝90°,
Figure PCTCN2020108314-appb-000070
Figure PCTCN2020108314-appb-000071
Z＝4。计算密度Dc＝1.374g/cm 3，单胞中电子数F(000)＝1184.0，单胞的线性吸收系数μ(Cu Kα)＝1.613mm –1，衍射试验温度T＝99.99(11)K。
11-P4S分子结构椭球图见图1，化合物结构为：
Figure PCTCN2020108314-appb-000072
步骤4：化合物11-P4S晶型A的表征
4.1 XPRD表征
4.1.1表征方法：XRPD在PANalytacal生产的X射线粉末衍射分析仪上采集，扫描参数如下表11所示。
表11 XRPD扫描参数
Figure PCTCN2020108314-appb-000073
4.1.1结果：
XPRD图谱见图2-1，图谱解析数据见表12。
表12 化合物11-P4S晶型A的XRPD图谱解析数据
序号 2θ±0.2(°) 相对强度(％) 序号 2θ±0.2(°) 相对强度(％)
1 6.33 100.00 17 23.83 3.97
2 8.34 3.02 18 24.60 15.50
3 10.87 27.37 19 24.77 14.22
4 13.77 14.23 20 25.39 3.68
5 14.42 6.81 21 26.02 1.63
6 15.83 7.24 22 26.45 5.28
7 16.61 10.15 23 27.20 7.57
8 17.46 1.17 24 27.66 1.06
9 18.20 15.22 25 28.15 0.71
10 18.43 3.93 26 29.16 0.83
11 18.89 25.20 27 29.90 0.69
12 19.27 17.88 28 30.72 0.52
13 20.05 10.97 29 31.90 2.72
14 20.51 4.31 30 33.76 0.98
15 22.19 17.13 31 38.28 0.93
16 22.77 6.84 32 39.21 0.85
4.2 TGA/DSC表征
4.2.1表征方法：TGA和DSC图分别在TA Q5000/5500热重分析仪和TA 2500差示扫描量热仪上采集，测试参数见表13。
表13 TGA/DSC测试参数
参数 TGA DSC
方法 线性升温 线性升温
样品盘 铝盘，敞开 铝盘，压盖/不压盖
温度范围 室温-设置终点温度 25℃-设置终点温度
扫描速率(℃/min) 10 10
保护气体 氮气 氮气
4.2.2结果：化合物11-P4S晶型A的TGA/DSC谱图见图2-2，结果显示样品加热至200℃后失重1.6％，并在215.4℃和246.2℃(峰值温度)有2个吸热峰。
4.3  1H NMR
4.3.1方法：液态核磁谱图在Bruker 400M核磁共振仪上采集，DMSO-d6作为溶剂。
4.3.2结果： 1H NMR图谱见图2-3，结果显示样品中对甲苯磺酸与游离碱摩尔比为1.0:1.0，MTBE与游离碱摩尔比为0.02:1.0，对甲苯磺酸质量分数为30.7％，MTBE质量分数为0.3％。
II.化合物11-P4S晶型B
步骤1.制备方法
将109mg化合物11-P4S晶型A溶于2毫升MeOH中，再加入18毫升THF反溶剂，置于-20℃下搅拌后过滤分离固体，将固体置于室温晾干得到化合物11-P4S晶型B。
步骤2.晶体表征
2.1表征方法：XPRD、TGA/DSC和 1H NMR的表征方法与化合物11-P4S晶型A的表征方法相同。
2.2试验结果
2.2.1 XPRD
XPRD谱图见图3-1，图谱解析数据见表14。
表14 化合物11-P4S晶型B的XRPD图谱解析数据
序号 2θ±0.2(°) 相对强度(％) 序号 2θ±0.2(°) 相对强度(％)
1 3.19 6.64 13 18.88 9.47
2 5.42 37.46 14 21.77 2.20
3 6.32 100.00 15 22.02 7.10
4 7.63 2.00 16 22.72 1.54
5 8.30 1.98 17 23.80 1.80
6 10.85 25.02 18 24.36 0.70
7 11.24 2.77 19 24.80 1.56
8 13.77 2.83 20 25.45 1.03
9 14.42 1.99 21 28.05 0.67
10 16.60 6.39 22 31.93 0.92
11 17.49 1.19 23 32.90 0.36
12 18.33 3.25 24 39.15 0.23
2.2.2 TGA/DSC
化合物11-P4S晶型B的TGA/DSC谱图见图3-2，其显示样品加热至200℃时有6.8％的失重，在120.5℃、221.8℃和252.6℃(峰值温度)有3个吸热峰。
2.2.3  1H NMR
1H NMR图谱见图3-3，其显示样品对甲苯磺酸与游离碱摩尔比为1.0:1.0，THF与游离碱的摩尔比为0.5，对应失重6.5％，未检测到甲醇残留。
III.化合物11-P4S晶型C
步骤1.制备方法
称取121.2mg化合物11-P4S晶型A，溶于2.2毫升MeOH中，再加入75毫升DCM澄清，置于室温搅拌两小时后仍澄清。转移至室温晾干得到化合物11-P4S晶型C。
步骤2.晶体表征
2.1表征方法：XPRD、TGA/DSC和 1H NMR的表征方法与化合物11-P4S晶型A的表征方法相同。
2.2试验结果
2.2.1 XPRD
XPRD图谱见图4-1，图谱解析数据见表15。
表15 化合物11-P4S晶型C的XRPD图谱解析数据
序号 2θ±0.2(°) 相对强度(％) 序号 2θ±0.2(°) 相对强度(％)
1 5.81 100.00 17 22.07 6.12
2 6.33 68.41 18 23.17 11.63
3 7.99 11.46 19 24.00 11.55
4 10.31 7.94 20 24.60 3.66
5 10.92 3.63 21 25.09 4.43
6 11.63 8.11 22 25.47 5.47
7 12.86 18.29 23 25.87 4.49
8 13.80 3.36 24 26.35 2.94
9 16.21 2.98 25 27.32 9.04
10 16.64 2.38 26 27.92 4.08
11 17.47 4.50 27 29.30 4.01
12 18.16 6.21 28 32.02 1.81
13 19.09 13.10 29 32.79 1.68
14 19.94 4.74 30 35.36 2.18
15 20.26 7.84 31 38.21 1.53
16 21.02 7.77      
2.2.2 TGA/DSC
化合物11-P4S晶型C的TGA/DSC谱图见图4-2，其显示样品加热至200℃时有17.4％的失重，并在112.5℃、210.7℃和249.6℃有3个吸热峰(峰值温度)。
2.2.3  1H NMR
1H NMR图见图4-3，其显示样品中对甲苯磺酸与游离碱摩尔比为1.0:1.0，DCM与游离碱的摩尔比为0.2，溶剂质量分数为3.1％，未检测到甲醇残留。
IV.化合物11-P4S晶型D
步骤1.制备方法
称取93.3mg化合物11-P4S晶型A，向其中加入3毫升1,4-dioxane，置于室温搅拌四天后，将样品抽滤，滤饼置于150℃加热约5分钟后得到化合物11-P4S晶型D。
步骤2.晶体表征
2.1表征方法：XPRD、TGA/DSC和 1H NMR的表征方法与化合物11-P4S晶型A的表征方法相同。
2.2试验结果
2.2.1 XPRD
XPRD图谱见图5-1，图谱解析数据见表16。
表16 化合物11-P4S晶型D的XRPD图谱解析数据
序号 2θ±0.2(°) 相对强度(％) 序号 2θ±0.2(°) 相对强度(％)
1 5.31 5.21 15 19.43 0.94
2 6.02 100.00 16 20.62 0.49
3 8.11 0.48 17 21.37 0.62
4 10.69 0.83 18 22.12 5.00
5 11.43 2.10 19 23.91 7.66
6 12.01 0.40 20 24.81 0.76
7 13.24 1.55 21 26.19 1.09
8 14.29 1.18 22 26.68 0.86
9 15.17 0.12 23 28.43 0.24
10 16.83 1.19 24 29.52 0.50
11 17.49 0.49 25 30.42 2.74
12 18.13 2.08 26 35.36 0.21
13 18.74 4.27 27 36.77 0.32
14 18.88 4.97      
2.2.2 TGA/DSC
化合物11-P4S晶型D的TGA/DSC谱图见图5-2，其显示样品加热至200℃时有1.3％的失重，在249.9℃有1个吸热峰(峰值温度)，并在188.0℃有1个放热峰(峰值温度)。
2.2.3 1H NMR
1H NMR见图5-3，其显示样品对甲苯磺酸与游离碱摩尔比为1.0:1.0，未检测到1,4-dioxane残留。
V.化合物11-P4S晶型E
步骤1.制备方法
称取62.1mg化合物11-P4S晶型A，向其中加入10毫升IPA，置于50℃室温搅拌2小时后，滤液缓慢降温(50℃～5℃,0.1℃/min)，析出适量固体后抽滤得到化合物11-P4S晶型E。
步骤2.晶体表征
2.1表征方法：XPRD、TGA/DSC和 1H NMR的表征方法与化合物11-P4S晶型A的表征方法相同。
2.2试验结果
2.2.1 XPRD
XPRD图谱见图6-1，图谱解析数据见表17。
表17 化合物11-P4S晶型E的XRPD图谱解析数据
序号 2θ±0.2(°) 相对强度(％) 序号 2θ±0.2(°) 相对强度(％)
1 5.24 1.72 11 18.93 0.30
2 5.46 0.28 12 22.15 0.33
3 6.06 100.00 13 22.71 0.12
4 6.25 12.62 14 24.50 1.68
5 8.26 0.25 15 25.17 0.31
6 10.56 0.29 16 25.33 0.28
7 12.17 2.33 17 30.79 5.93
8 13.73 0.13 18 30.88 2.88
9 15.87 0.33 19 31.83 0.25
10 18.32 6.29      
2.2.2 TGA/DSC
化合物11-P4S晶型E的TGA/DSC谱图见图6-2，其显示样品加热至200℃时有2.8％的失重，并在244.6℃有1个吸热峰(峰值温度)。
2.2.3  1H NMR
1H NMR图见图6-3，其显示示样品对甲苯磺酸与游离碱摩尔比为1.0:1.0，未检测到MTBE残留。
VI.化合物11-P3S晶型
步骤1：化合物11-P3成单对甲苯磺酸盐：
Figure PCTCN2020108314-appb-000074
将11-P3(0.20g，0.52mmol)溶于乙酸乙酯(5ml)中，滴加对甲苯磺酸一水合物(0.12g,0.62mmol)的乙酸乙酯溶液，析出白色固体。室温搅拌12h，抽滤，滤饼用乙酸乙酯洗涤(5mL*3)，滤饼干燥，得化合物11-P3S白色固体0.22g，收率78％。
步骤2：化合物11-P3S的单晶培养方法：
1)称取11.6mg的11-P3S于1.5mL HPLC小瓶中。
2)向固体中加入348微升的乙醇，升温至60度，并在60度下恒温直至固体全部溶清。
3)静置状态下将溶液以0.5℃/min降至25度。
4)析出晶体，将反应瓶拿到显微镜下观察，晶体合格，进行XRSD试验。
步骤3：11-P3S的单晶XRSD试验：
3.1仪器参数与数据收集
3.1.1仪器参数：与化合物11-P4S晶型A的3.1.1部分相同
3.1.2数据收集：衍射实验收集了36655个衍射点，其中独立衍射点4793个(R int＝0.0525)。衍射收集范围2θ＝6.342 to 133.17°，衍射指标范围-32≤h≤30,-19≤k≤16,-7≤l≤7。结构解析使用SHELXT(Sheldrick,G.M.2015.ActaCryst.A71,3-8)，结构精修使用SHELXL(against F 2)(Sheldrick,G.M.2015.ActaCryst.C71,3-8)。4793个独立衍射点中，参加结构精修的参数为348。精修后S＝1.041,R 1＝0.0324,wR 2＝0.0824。残余电子密度值为0.32和
Figure PCTCN2020108314-appb-000075
3.2数据列表见表18和表19
表18：化合物11-P3S的单晶衍射数据列表
Figure PCTCN2020108314-appb-000076
Figure PCTCN2020108314-appb-000077
表19：原子坐标(x 10^4)与等效各向同性位移参数(A^2 x 10^3)
Figure PCTCN2020108314-appb-000078
3.2结论
化合物11-P3S晶型为无色块状(0.20×0.20×0.10mm 3)，属于正交晶系P21212空间群。晶胞参数：
Figure PCTCN2020108314-appb-000079
α＝90°,β＝90°,γ＝90°,
Figure PCTCN2020108314-appb-000080
Figure PCTCN2020108314-appb-000081
Z＝4。计算密度Dc＝1.375g/cm 3，单胞中电子数F(000)＝1184.0，单胞的线性吸收系数μ(Cu Kα)＝1.614mm –1，衍射试验温度T＝99.99(11)K。化合物11-P3S分子结构椭球图见图7，化合物结构为：
Figure PCTCN2020108314-appb-000082
实施例31
Figure PCTCN2020108314-appb-000083
氮气保护，冰浴下，将Boc-L-缬氨酸(857.6mg，3.95mmol)溶于20mL二氯甲烷中，加入4-二甲氨基吡啶(386.9mg，3.16mmol)，化合物11-P4(1.02g，2.63mmol)，冰浴下搅拌反应5分钟。一次性加入二环己基碳二亚胺(813.7mg，3.95mmol)，自然升温，反应18小时。过滤，滤液减压浓缩，用柱层析分离(二氯甲烷：甲醇＝30:1)纯化所得残余物，得化合物Em1-11P4a(1.13g，类白色固体)，收率：73.2％。MS m/z(ESI)：587.3[M+1]
将化合物Em1-11P4a(1.13g，1.92mmol)，溶于15ml浓度为4M的HCl的1,4-二氧六环溶液中，室温下反应2小时。浓缩反应液，乙醚(10ml*1)洗涤固体，得粗品。将粗品溶于30mL水中，饱和碳酸氢钠水溶液调Ph＝7～8,二氯甲烷(10ml*3)萃取，合并有机相。用水(10ml*1)和饱和氯化钠(10ml*1)分别洗涤二氯甲烷相。无水硫酸钠干燥二氯甲烷相。过滤，滤去固体，浓缩二氯甲烷相，得化合物21-P3(720mg)，收率：76.8％。MS m/z(ESI)：487.3[M+H] +。
实施例32
Figure PCTCN2020108314-appb-000084
取7.53g化合物19，采用HPLC法，使用大赛璐的制备色谱和大赛璐手性柱对手性异构体分离，收集其相应组分，旋转蒸发除去溶剂，得到光学异构体的纯品。分离方法参见表20。
表20：化合物19的手性分离方法
Figure PCTCN2020108314-appb-000085
Figure PCTCN2020108314-appb-000086
表21：化合物19的手性分析结果
峰号 保留时间 峰面积 相对峰面积％
1 3.481 246012 14.159
2 5.397 1225940 70.559
3 12.004 265519 15.282
分别收集19P2(保留时间5.397min)和其他组分，得到5.11g化合物19P2，产品纯度98.15％。MS m/z(ESI)：360.2[M+H] +； 1H NMR(600MHz,CDCl 3):δ6.63(s,1H),6.56(s,1H),3.83-3.79(m,5H),3.49-3.46(m,1H),3.42-3.35(m,1H),3.28-3.02(m,4H),2.89(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.33(t,1H),1.82-1.75(m,1H),1.70-1.58(m,1H),1.06-0.99(m,1H),0.93-0.87(m,6H),0.64-0.59(m,2H),0.36-0.31(m,2H).
实施例33
步骤1：化合物19P2成单对甲苯磺酸盐：
Figure PCTCN2020108314-appb-000087
将化合物19P2(3.00g，8.34mmol)溶于乙酸乙酯(30ml)中，滴加对甲苯磺酸一水合物(2.37g,12.5mmol)的乙酸乙酯溶液。室温搅拌12h，抽滤，滤饼用乙酸乙酯(10mL*3)洗涤。收集滤饼，干燥，得白色固体3.59g，收率81％。
步骤2：化合物19P2S单晶培养
称取10mg的19P2S于1.5mL HPLC小瓶中，向固体中加入500微升的乙醇，升温至40度，并在40度下恒温直至固体全部溶清，静置状态下将溶液以0.3℃/min降至25度，析出晶体，将反应瓶拿到显微镜下观察，晶体合格，进行XRSD试验。
步骤3：19P2S的单晶XRSD试验：
(1)仪器参数：与化合物11-P4S晶型A的3.1.1部分相同
(2)数据收集
衍射试验收集了24167个衍射点，其中独立衍射点4899个(Rint＝0.0646)。衍射收集范围2θ＝6.33 to 133.182°，衍射指标范围-7≤h≤5,-18≤k≤17,-33≤l≤33。结构解析使用SHELXT (Sheldrick,G.M.2015.ActaCryst.A71,3-8)，结构精修使用SHELXL(against F 2)(Sheldrick,G.M.2015.ActaCryst.C71,3-8)。4899个独立衍射点中，参加结构精修的参数为339。精修后S＝1.020,R 1＝0.0373,wR 2＝0.0920。残余电子密度值为0.38和
Figure PCTCN2020108314-appb-000088
(3)数据列表见表22和表23
表22 化合物19P2S单晶衍射数据列表
Figure PCTCN2020108314-appb-000089
表23：原子坐标(x 10^4)与等效各向同性位移参数((A^2 x 10^3)
Figure PCTCN2020108314-appb-000090
Figure PCTCN2020108314-appb-000091
结论：化合物19P2S晶体为无色块状(0.30×0.10×0.04mm3)，属于正交晶系P212121空间群。晶胞参数
Figure PCTCN2020108314-appb-000092
α＝90°,β＝90°,γ＝90°,
Figure PCTCN2020108314-appb-000093
Z＝4。计算密度Dc＝1.273g/cm 3，单胞中电子数F(000)＝1144.0，单胞的线性吸收系数μ(Cu Kα)＝1.385mm –1，衍射试验温度T＝100.00(13)K。
化合物19P2S的分子结构椭球图见图8。化合物的结构为：
Figure PCTCN2020108314-appb-000094
实施例34
Figure PCTCN2020108314-appb-000095
将Boc-L-缬氨酸(260mg，1.2mmol)溶于10mL二氯甲烷中，在0℃下，加入二环己基碳二亚胺(309mg，1.5mmol)、4-二甲氨基吡啶(12.2mg，0.1mmol)，然后加入3-异丁基-10-甲氧基-9-环丙甲基-2,3,4,6,7,11b-六氢-1H-吡啶[2,1-a]异喹啉-2-醇(359mg，1.0mmol)，升至室温搅拌反应过夜。加入20mL水洗涤，分液，将有机相浓缩后加入6ml浓度为4M的盐酸 的二氧六环溶液，室温搅拌2h，减压蒸出溶剂，加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(10mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩得目标化合物(298mg，淡黄色油状液体)。MS m/z(ESI)：459.3[M+H] +； 1H NMR(600MHz,CDCl 3):δ6.71(s,1H),6.62(s,1H),4.76-4.70(m,1H),3.83-3.79(m,5H),3.48-3.44(m,1H),3.42-3.35(m,1H),3.28-3.02(m,4H),2.89(dd,1H),2.77-2.61(m,2H),2.60-2.48(m,2H),2.36-2.30(m,1H),1.88-1.75(m,2H),1.70-1.58(m,3H),1.06-0.99(m,1H),0.95-0.84(m,12H),0.65-0.59(m,2H),0.37-0.33(m,2H).
对照例35：
Figure PCTCN2020108314-appb-000096
合成路线：
Figure PCTCN2020108314-appb-000097
将3-异丁基-9,10-二羟基-1,3,4,6,7,11b-六氢-2H-吡啶并[2,1-a]异喹啉-2-酮(1.7g，5.89mmol)溶解于30mL乙腈中，加入碳酸铯(9.6g，5mmol)，溴甲基环丙烷(1.91g，14.1mmol)升温至80℃，搅拌反应5小时。反应液浓缩，用薄层色谱法(石油醚：乙酸乙酯＝5:1)纯化所得残余物，得对照例35化合物。
对照例36：
Figure PCTCN2020108314-appb-000098
合成路线：
Figure PCTCN2020108314-appb-000099
将3-异丁基-9,10-二羟基-1,3,4,6,7,11b-六氢-2H-吡啶并[2,1-a]异喹啉-2-酮(1.00g,3.45mmol)溶解于50mL N,N-二甲基甲酰胺中，加入碳酸钾(2.38g,17.2mmol)，搅拌30分钟，加入三氟溴丙烷(1.46g,10.4mmol)，氮气保护，升温至80℃，搅拌反应8小时。反应液降至室温加入50mL水淬灭反应，加入乙酸乙酯(50mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，硅胶柱层析(乙酸乙酯：石油醚＝1：5)，得对照例36化合物(0.80g,淡黄色固体)。
对照例37：
Figure PCTCN2020108314-appb-000100
合成路线：
Figure PCTCN2020108314-appb-000101
1、将3-异丁基-9,10-二羟基-1,3,4,6,7,11b-六氢-2H-吡啶并[2,1-a]异喹啉-2-酮(0.50g,1.73mmol)溶解于5mL N,N-二甲基甲酰胺中，加入碳酸钾(0.24g,1.73mmol)，搅拌30分钟，加入3-氟溴丙烷(0.24g,1.73mmol)，氮气保护，升温至80℃，搅拌反应8小时。反应液降至室温加入20mL水淬灭反应，加入乙酸乙酯(30mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，硅胶柱层析(乙酸乙酯：石油醚＝1:10)，得3-异丁基-9-3‘-氟丙氧基-10-羟基-1,3,4,6,7,11b-六氢-2H-吡啶并[2,1-a]异喹啉-2-酮(0.42g,无色液体)。
2、将3-异丁基-9-3‘-氟丙氧基-10-羟基-1,3,4,6,7,11b-六氢-2H-吡啶并[2,1-a]异喹啉-2-酮(0.07g,0.20mmol)溶解于5mL N,N-二甲基甲酰胺中，加入碳酸钾(0.06g,0.40mmol)，搅拌30分钟，加入1-溴丙烷(0.037g,0.30mmol)，氮气保护，升温至80℃，搅拌反应8小时。反应液降至室温加入20mL水淬灭反应，加入乙酸乙酯(30mL×3)萃取，合并有机相，无水硫酸钠干燥，过滤，滤液减压浓缩，硅胶柱层析(EA：PE＝1：5)，得对照例37化合物(0.04g,白色固体)。MS m/z(ESI):392.3[M+H] +；1H-NMR(600MHz,CDCl 3)δ6.67(s,1H),6.61(s,1H),4.77-4.69(t,J＝5.8Hz,1H),4.68-4.60(t,J＝5.8Hz,1H),4.17-4.05(m,2H),3.84-3.73(m,2H),3.54-3.38(m,1H),3.34-3.23(m,1H),3.17-2.93(m,2H),2.93-2.83(m,1H),2.78-2.67(m,2H),2.66-2.48(m,2H),2.35(t,J＝11.6Hz,1H),2.25-2.13(m,2H),1.86-1.74(m,1H),1.73-1.61(d,J＝6.4Hz,1H),1.32-1.20(m,1H),1.08-0.98(m,1H),0.97-0.84(m,6H),0.39-0.29(m,3H).
对照例38：
第一步：片段2合成
Figure PCTCN2020108314-appb-000102
Figure PCTCN2020108314-appb-000103
1、50g化合物g溶于150毫升DMF中，加入苄基溴57.3g，碳酸钾68.2g，氮气保护条件下，室温反应6h，TLC检测发现原料消失。体系倒入冰水，乙酸乙酯萃取3次，合并有机相用盐水洗涤两次，有机相无水硫酸钠干燥，过滤，浓缩，得67.87g淡黄色固体产品h，直接用于下一步。
2、67.78g化合物h，加入硝基甲烷100毫升，乙酸铵13.9g，加热至112度反应4h，TLC检测发现原料消失。降温，蒸掉硝基甲烷，残余物用水洗，乙酸乙酯萃取2次，有机相无水硫酸钠干燥，过滤，浓缩，得77g黄色固体i。用于下一步。
3、30g氢化铝锂溶于300毫升THF中，降温至0度，将77g化合物i溶于四氢呋喃中，缓慢滴入反应瓶中，滴毕，72度回流反应3h。TLC检测发现原料消失。降温至零度，缓慢依次加入30毫升水，60毫升10％的氢氧化钠溶液，90毫升水，混合物抽滤，滤饼用四氢呋喃洗涤两次，合并有机相。蒸干得79.8g褐色油状液体。用丙酮溶解，加入草酸，调pH＝3析出固体，抽滤得40g黄白色固体化合物j。用于下一步。
4、2.9g化合物j溶于30毫升乙酸中，加入10毫升三氟乙酸，乌洛托品3.3g，加热至85度反应4h，TLC检测发现原料消失。降温，蒸掉乙酸和三氟乙酸，残余物加水，用氢氧化钠水溶液调pH＝9，用乙酸乙酯萃取3次，有机相无水硫酸钠干燥，过滤，浓缩，得2.9g褐色油状液体k，直接用于下一步。
5、39.2g化合物k溶于100毫升乙醇和100毫升水混合溶液中，加入27.6g 3-二甲基氨基-5-甲基-2-己酮，苄基三乙基氯化铵10.1g，加热至95度反应16h，TLC检测发现原料消失。降温，蒸掉乙醇，残余物用乙酸乙酯萃取3次，有机相无水硫酸钠干燥，过滤，浓缩，得产品34.4为褐色油状液体，用丙酮溶解，加入对甲苯磺酸，调pH＝3析出固体，抽滤得12.7g黄白色固体l，用于下一步。
6、1.5g化合物l溶于20毫升甲醇溶液中，加入两小勺钯碳，在氢气条件下常温反应10h，TLC检测发现原料消失。反应物抽滤，滤饼用甲醇洗涤两次，合并有机相。蒸干得1.08g淡黄色固体片段2。
第二步：
Figure PCTCN2020108314-appb-000104
合成路线：
Figure PCTCN2020108314-appb-000105
0.2g片段2溶于5毫升DMF中，加入1-溴丙烷0.08g，碳酸钾0.14g，氮气保护条件下，加热至70度反应3h，TLC检测发现原料消失。降温，体系加入冰水，乙酸乙酯萃取3次，合并有机相用盐水洗涤两次，有机相无水硫酸钠干燥，过滤，浓缩，得0.15g白色固体产品对照例38化合物。
对照例39：
Figure PCTCN2020108314-appb-000106
合成路线：
Figure PCTCN2020108314-appb-000107
0.2g片段2化合物溶于5毫升DMF中，加入1-溴-3氟丙烷0.08g，碳酸钾0.14g，氮气保护条件下，加热至70度反应3h，TLC检测发现原料消失。降温，体系加入冰水，乙酸乙酯萃取3次，合并有机相用盐水洗涤两次，有机相无水硫酸钠干燥，过滤，浓缩，柱纯化，得产品0.15g为白色固体，收率84.5％，MS(ESI):364[M+H] +；1H NMR(400MHz,CDCl 3)6.63(s,1H),6.60(s,1H),4.68-4.70(m,1H),4.60-4.62(m,1H)，4.08-4.10(m,2H),3.84(s,3H),3.83-3.84(m,1H),3.48-3.50(m,1H),3.28-3.29(m,2H),2.87-2.89(m,1H),2.52-2.75(m,4H),2.34-3.35(m,1H),2.21-2.23(m,2H),1.78-1.82(m,1H),1.65-1.67(m,1H),1.25-1.28(m,1H),1.01-1.04(m,1H),0.87-0.93(m,6H).
对照例40：
Figure PCTCN2020108314-appb-000108
合成路线：
Figure PCTCN2020108314-appb-000109
片段2化合物0.2g溶于5毫升DMF中，加入溴甲基环丙烷0.08g，碳酸钾0.14g，氮气保护条件下，加热至70度反应3h，TLC检测发现原料消失。降温，体系加入冰水，乙酸乙酯萃取3次，合并有机相用盐水洗涤两次，有机相无水硫酸钠干燥，过滤，浓缩，得0.15g对照例40化合物，为白色固体。MS m/z(ESI):358.3[M+H] +；1HNMR(400MHz,CDCl 3)6.62(s,1H),6.57(s,1H)，3.85(s,3H),3.78-3.79(，2H),3.48-3.50(m,1H),3.28-3.29(m,1H),3.11-3.14(m,2H),2.86-2.89(m,1H),2.52-2.75(m,4H),2.35(m,1H),1.78-1.80(m,1H),1.26-1.31(m,2H),1.02-1.05(m,1H),0.87-0.93(m,6H),0.61-0.64(m,2H)，0.32-0.35(m,2H).
对照例41：
第一步：片段3合成
Figure PCTCN2020108314-appb-000110
合成路线：
Figure PCTCN2020108314-appb-000111
1、化合物3a(10.0g,72.5mmol)溶于100mL DMF中，加入碳酸钾(15.0g,109mmol)与溴苄(18.6g,109mmol)，升温至85℃，反应8h。冷却至室温，将反应液加入500mL冰水析出固体，过滤，干燥，得化合物3b(9.92g,白色固体)。
2、化合物3b(9.92g，43.5mmol)溶于50mL DMF中，加入溴乙烷(7.11g，65.2mmol)， 碳酸钾(9.00g，65.2mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入500mL水中，然后用乙酸乙酯萃取(200mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用石油醚(150mL)打浆残留物。过滤，收集固体，得化合物3c(10.0g，类白色固体)。
3、化合物3c(10.0g，39.1mmol)溶于50mL硝基甲烷，加入乙酸铵(1.81g，23.5mmol)加热至115℃，反应3h。冷却后，浓缩，残余物用水洗，乙酸乙酯(100mL*3)萃取，合并有机相，有机相无水硫酸钠干燥，过滤，浓缩，得化合物3d(11.6g，黄色固体)。
4、氮气保护，冰浴下，将化合物3d(11.6g，38.8mmol)溶于100mL无水THF中，滴加至氢化铝锂(4.42g，116mmol)的100mL无水THF中，反应1h。升温至60℃，反应2h。冰浴降温后，滴加4.4mL水，然后加入8.8mL 10％氢氧化钠溶液与13.2mL水。抽滤，然后用乙酸乙酯洗涤(150mL*3)滤饼，滤液减压浓缩。浓缩液用乙酸乙酯稀释，加入草酸的乙酸乙酯溶液，直至pH值显酸性，搅拌过夜。过滤，收集滤饼，用乙酸乙酯洗涤(100mL*3)滤饼，得化合物3e(11.2g，白色固体)。
5、将化合物3e(11.2g，31.0mmol)溶于100mL乙酸中，加入30mL三氟乙酸中，加入乌洛托品(9.55g，68.2mmol)，加热至85℃反应4h。冷却至室温浓缩，残余物加水，用氢氧化钠水溶液调pH＝9，用乙酸乙酯(100mL*3)萃取，合并有机相，有机相无水硫酸钠干燥，过滤，浓缩，得化合物3f(8.71g，褐色油状物，粗品)，未经纯化，直接用于下一步。
MS m/z(ESI)：282.2[M+H] +
6、将化合物3f(8.71g，31.0mmol)溶于100mL乙醇和100mL水混合溶液中，加入3-二甲基氨基-5-甲基-2-己酮(6.36g，37.2mmol)，苄基三乙基氯化铵(2.12g，9.30mol)加热至95℃反应18h。冷却至室温浓缩，残余物用乙酸乙酯(150mL*3)萃取，合并有机相，有机相无水硫酸钠干燥，过滤，减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝5:1)，得化合物3g(3.78g，类白色固体)。MS m/z(ESI)：408.3[M+H] +
7、将化合物3g(3.78g，9.30mmol)溶于50mL甲醇溶液中，加入10％的有水钯碳(0.5g)，通入氢气，室温下反应18h。抽滤，滤饼用甲醇(50mL*2)洗涤，减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝2:1)，得片段3(2.50g，类白色固体)。MS m/z(ESI)：318.2[M+H] +
第二步：
Figure PCTCN2020108314-appb-000112
合成路线：
Figure PCTCN2020108314-appb-000113
1、将片段3化合物(200mg，0.631mmol)溶于5mL DMF中，加入溴丙烷(116mg，0.946mmol)、碳酸钾(130mg，0.946mmol)，升温至80℃，反应5h。冷却后，将反应体系倾入30mL水中，然后用乙酸乙酯萃取(20mL*3)，合并有机相，用无水硫酸钠干燥，过滤，滤液减压浓缩，用柱层析分离(石油醚：乙酸乙酯＝4:1)，得对照例41化合物(182mg，类白色固体)。MS m/z(ESI)：360.2[M+1]
对照例42：
Figure PCTCN2020108314-appb-000114
合成路线：
Figure PCTCN2020108314-appb-000115
将对照例36化合物(0.80g，2.00mmol)溶解于30mL无水乙醇中，0℃下分批加入硼氢化钠(0.15g，4.00mmol)，0℃搅拌反应2小时。加入6mL饱和氯化铵溶液淬灭反应，反应液过滤，滤液减压浓缩，硅胶柱层析(二氯甲烷：甲醇＝10：1)，得0.75g淡黄色固体化合物。MS m/z(ESI):412.3[M+H] +；1HNMR(600MHz,CD 3OD)δ6.82(s,1H),6.72(s,1H),4.71-4.63(m,2H),4.63-4.53(m,2H),4.11-4.01(m,4H),3.23-3.14(m,1H),3.11-2.99(m,3H),2.73-2.64(m,1H),2.60-2.52(m,1H),2.52-2.43(m,1H),2.19-2.06(m,4H),2.07-1.97(m,1H),1.78-1.61(m,3H),1.52-1.38(m,1H),1.08-0.99(m,1H),0.98-0.86(m,6H).
对照例43-44
对照例43-44的制备参考对照例42的方法，通过乙醇和硼氢化钠的溶液进行还原制得。
表24：对照例43-44结构和表征数据
Figure PCTCN2020108314-appb-000116
Figure PCTCN2020108314-appb-000117
对照例45：
Figure PCTCN2020108314-appb-000118
合成路线：
Figure PCTCN2020108314-appb-000119
1、氮气保护，冰浴下，将Boc-L-缬氨酸(651mg，3mmol)溶于15mL二氯甲烷中，加入4-二甲氨基吡啶(293mg，2.4mmol)，对照例7化合物(760mg，1.83mmol)，冰浴下搅拌反应5分钟。一次性加入二环己基碳二亚胺(618mg，3mmol)，自然升温，反应18小时。过滤，滤液减压浓缩，用柱层析分离(二氯甲烷：甲醇＝30:1)纯化所得残余物，得中间化合物(780mg，类白色固体)。
MS m/z(ESI)：565.4[M+1]
2、将上一步得到的化合物(780mg，1.38mmol)，溶于10ml二氯甲烷中，加入1.7ml浓度为4M的1,4-二氧六环溶液中，室温下反应2小时。浓缩反应液，乙醚(10ml*1)洗涤固体，得粗品。将粗品溶于30mL水中，饱和碳酸氢钠水溶液调Ph＝7～8,二氯甲烷(10ml*3)萃取，合并有机相。水(10ml*1)和饱和氯化钠(10ml*1)分别洗涤二氯甲烷相。无水硫酸钠干燥二氯甲烷相。过滤，滤去固体，浓缩二氯甲烷相，得对照例45化合物(500mg，类白色固体)。MS m/z(ESI)：465.4[M+H] +； 1H NMR(400MHz,CD 3OD)δ6.73-6.75(m,2H),4.56-4.77(m,3H),4.08-4.11(m,2H),3.80(s,3H),3.05-3.32(m,4H),2.50-2.75(m,3H),2.01-2.21(m,5H),1.33-1.77(m,3H),1.06-1.13(m,2H),0.93-1.05(m,12H).
试验例1 生物活性测试
一、化合物与大鼠VMAT2结合活性的放射性检测(Binding assay)
1.试验目的：
测定各化合物与大鼠VMAT2结合的IC 50和Ki值，评价化合物与VMAT2的亲和力。
2.试验材料
配体：[3H]Dihydrotetrabenazine(DHTBZ)(10nM)
受试化合物：
化合物1-9,11-12,14,16-19,20,27-28,32,11-P3,11-P4,21-P3：按照上述对应实施例制备得到
对照例35-43化合物：按照对照例35-43制备得到
TBZ：江苏威凯尔医药科技有有限公司，批号TBZ-113030
DHTBZ：江苏威凯尔医药科技有有限公司，批号67-25-1521-59C
DHTBZ-X(消旋体)：以TBZ为原料，按照WO2008058261的Reaction Scheme 1制备
VBZ：按照以下方法制备：将VBZ二甲苯磺酸盐(0.5g，0.65mmol)溶于10mL水中，用饱和NaHCO3溶液调pH＝8左右，用EA(20mL*3)萃取，合并有机相，无水硫酸纳干燥，浓缩，得VBZ 0.26g白色固体。
3.试验步骤和方法
3.1大鼠脑泡膜制备
选取体重175±25g的雄性Wistar大鼠，手术分离出全脑(不包括小脑)，置于20毫升预冷的0.32M蔗糖溶液中，用Teflon研杵匀浆器进行匀浆。匀浆物经4℃，1000g离心12min；吸取上清液，4℃继续22,000g离心10min，弃上清，将所得沉淀置于18mL冰冷的MilliQ水(Millipore Corporation，Billerica，MA)中孵育5分钟，进行渗透压休克以破碎细胞膜。然后再加入2mL HEPES(25mM)溶液和2mL酒石酸钾溶液(100mM)恢复渗透压。将所得样品4℃20,000g，离心20min，吸取上清液，加入20μL MgSO4(1mM)溶液。将溶液4℃，100,000g离心45min，收集沉淀，重悬于冰冷的测定缓冲液中(25mM HEPES，100mM酒石酸钾，5mM MgSO4，0.1mM EDTA和0.05mM EGTA，pH7.5)，获得囊泡悬液。
3.2检测分析
采用96孔板检测，设2-3个复孔。将50μL囊泡悬液(含32μg蛋白)与10nM[3H]Dihydrotetrabenazine(DHTBZ)，以及50μL含受试化合物的溶液(抑制剂浓度范围在1nM-1000nM，或其他所需浓度)，加入96孔板的每孔中，于25℃下孵育30min。用非特异性配体Ro4-1284(10μM)确定预测非特异性结合，以有明确药理学特征的丁苯那嗪(Tetrabenazine，TBZ)、DHTBZ、VBZ或者DHTBZ消旋体作为阳性对照，并用于与新化合物活性的比较。孵育结束后，对反应液进行过滤(滤菌收集器，PerkinElmer Life和Analytical Sciences)至过滤板，随后用350μL冰冷的缓冲液(25mM HEPES，100mM酒石酸钾，5mM MgSO 4和10mM NaCl，pH 7.5)洗涤滤膜5次。过滤板干燥并密封底部，每个孔加满40μL闪烁混合物(MicroScint 20；PerkinElmer Life和Analytical Sciences)。通过液体闪烁光谱测定法(TopCount  NXT；PerkinElmer Life and Analytical Sciences)测定滤器上的放射性。
3.3结果分析
根据上述放射性测定结果计算IC 50和Ki。采用MathIQTM(ID Business Solutions Ltd.,UK).非线性、最小二乘法回归分析计算IC 50。Ki值采用Cheng和Prusoff(Cheng,Y.,Prusoff,W.H.,Biochem.Pharmacol.22:3099-3108,1973)公式，结合测试化合物的IC 50和Eurofins Panlabs公司的该放射性检测方法中历史KD计算Ki值。
采用上述方法测试部分实施例和对照例化合物的20nM和100nM的结合抑制率、IC 50值和Ki值结果见表25-27。
表25：本发明部分实施例化合物结合抑制率数据
化合物 20nM结合抑制率％ 100nM结合抑制率％ IC 50(nM)
TBZ 50 77 23
DHTBZ 64 82 10.9
实施例1 29 70 45
实施例2 74 87 6.27
实施例3 74 84 3.37
实施例4 87 93 0.98
实施例5 75 94 10.6
实施例6 77 100 5.13
实施例7 72 95 5.40
实施例8 43 85 23
实施例9 76 97 6.03
实施例16 65 84 12.9
实施例17 28 71 58
实施例19 74 94 5.99
实施例20 68 102 4.90
对照例35 15 36 190
对照例36 17 22 >500
对照例37 11 24 >500
对照例38 2 5 >500
对照例39 11 20 >500
对照例40 1 2 >500
对照例41 22 29 280
对照例42 11 3 >500
对照例43 6 22 >500
表26：本发明部分实施例化合物结合抑制率数据
化合物 IC 50 Ki
DHTBZ 7.23nM 4.22nM
VBZ >0.5μM 未测得
11 5.45nM 3.18nM
11-P3 0.47μM 0.27μM
11-P4 2.54nM 1.48nM
21-P3 0.18μM 0.11μM
表27：本发明部分实施例化合物结合抑制率数据
化合物 IC 50(nM) Ki(nM)
DHTBZ-X 21 12.3
11 6.13 3.57
12 3.78 2.21
14 10 5.83
18 3.55 2.07
19 2.30 1.34
27 2.06 1.20
28 7.70 4.49
32 1.29 0.75
试验结果显示：相对于TBZ、DHTBZ、DHTBZ–X、VBZ和对照例化合物，本发明提供的化合物与VMAT具有更强的亲和力。当10位为甲基，9位为乙基、环丙基亚甲基和4-氟丁基时，结合抑制活性最好；当10位为丙基、丁基等长取代基时，结合抑制活性明显的降低，甚至接近没有活性。而当9位为甲基时，10位为乙基、丙基、环丙基亚甲基等长取代基时活性明显降低甚至没有活性。也即是，本发明人发现9位和10位的取代基对活性有显著的差异。
二、化合物与VMAT2摄取试验(uptake assay)
1、目标
测试化合物VMAT2的uptake assay。
2.材料
(1)试剂和材料
3H-多巴胺、蔗糖、HEPES、酒石酸钾、EGTA、EDTA、ATP、MgCl2、MgSO4、抗坏血酸、TBZ、BCA蛋白质检测试剂盒
冷冻囊泡悬浮液：采用SD大鼠纹状体提取囊泡悬液。新鲜大鼠纹状体加入28mL冰浴的0.32M的蔗糖溶液，用匀浆仪进行匀浆10次，每次10s。4℃条件下，对匀浆液进行离心2000g，10min。吸取上清液，继续4℃，10000g离心30min。分离沉淀，重悬于4mL 0.32M的蔗糖溶液，再加入14mL冰浴MilliQ水，使渗透休克。1min后，加入1.8mL 0.25M HEPES缓冲液和1.8mL1M酒石酸钾溶液。4℃，20000g离心30min。收集上清，继续4℃，55000g离心60min。弃上清，加入200μL 10mM MgSO4，200μL 0.25M HEPES和200μL 1M酒石酸钾。4℃，55000g离心45min。收集沉淀，重选于10mL检测缓冲液(25mM HEPES，100mM酒石酸钾，50μM EGTA，100μM EDTA，20mM MgCl2，2mM ATP，pH7.4)，500μL/管分装，-80℃冻存备用。
(2)缓冲区
分析缓冲液：25mM HEPES，100mM酒石酸钾，50μMEGTA，100μMEDTA，20mM MgCl 2,1.7mM抗坏血酸，2mM ATP，pH7.4。在测定开始前加入抗坏血酸和ATP。
洗涤缓冲液：25mM HEPES，100mM酒石酸钾，50μM EGTA，100μM EDTA，pH 7.4。
(3)受试化合物
化合物11-15,18-20,26-28,32,11-P4,21-P3：按照上述对应的实施例制备得到
DHTBZ、DHTBZ-X、VBZ：来源与上述Binding试验中相同
(4)仪器和消耗品
Unifilter-96 GF/B滤板，Perkin Elmer(Cat#6005177)；
96孔锥形聚丙烯板，安捷伦(Cat#5042-1385)；
TopSeal-A密封膜，Perkin Elmer(Cat#6050185)；
MicroBeta2(PerkinElmer)；
细胞收获C961961，(Perkin Elmer)；
SpectraMax 340PC(Molecular Devices)；
3、试验方法
(1)用DMSO4倍稀释待测样品和TBZ至8个浓度梯度，最高浓度0.2mM，最低浓度1μM。用移液枪转移1μl稀释的待测化合物和TBZ至检测板。
(2)加1μl 2mM TBZ作为非特异性结合对照(LC)；1μl DMSO作为总结和对照(HC)。
(3)加100μl稀释过的囊泡悬液(含15μl储存液)至96-孔板，37℃孵育15min。
(4)用检测缓冲液稀释3H-Dopamine(17.92μM)至0.2μM.加100μl 0.2μM的3H-Dopamine至检测板，终浓度0.1μM，然后37℃孵育10min。
(5)将反应混合物过滤采集器的GF/B板，用预冷的漂洗缓冲液漂洗GF/B板4次。
(6)0℃干燥板至少1hr。
(7)干燥后，用Perkin Elmer Unifilter-96封底胶带密封滤板，加50μl Perkin Elmer Microscint 20混合液，用Perkin Elmer TopSeal-A封膜封住滤板顶部。
(8)用Perkin Elmer MicroBeta2 Reader计数滤膜捕获的3H数量。
(9)用Prism 5.0软件分析数据.采用“log(Inhibitor)vs.response-Variable Slope”模型计算IC 50，并采用方程ICanything＝IC 50*(anything/(100-anything))1/slope计算IC 90。
4、试验结果见表28-30。
表28：本发明部分实施例化合物uptake数据
化合物 IC 50(nM) IC 90(nM)
DHTBZ 12.72 148.96
11 6.04 45.97
12 16.92 67.56
13 5.13 110.3
15 4.29 48.93
20 5.71 23.21
表29：本发明部分实施例化合物uptake数据
化合物 IC 50(nM) IC 90(nM)
DHTBZ 30.61 276.77
VBZ 313.0 2916.82
11 7.891 36.68
11-P3 129.10 1598.59
11-P4 10.2 92.39
21-P3 230.2 1314.74
表30：本发明部分实施例化合物uptake数据
化合物 IC 50(nM) IC 90(nM)
DHTBZ-X 42.29 161.34
12 11.95 347.66
14 192.80 1480.14
18 33.62 187.87
19 4.207 92.33
26 14.39 140.82
27 8.99 331.52
28 5.93 116.78
32 2.100 4.37
试验结果表明，相对于DHTBZ、DHTBZ–X和VBZ，本发明提供的化合物具有更强的体外活性。
试验例2 化合物11、16、21、22、23和对照例44、45的SD大鼠药动学评价
1、试验材料
a)受试化合物
化合物11、16、21-23、对照例44、对照例45：按照相应实施例制备得到
DHTBZ：江苏威凯尔医药科技有有限公司，批号67-25-1521-59C
VBZ：按照试验例1中提及的方法制备
b)溶媒：20％solutol溶液，Solutol批号：BCBQ5646V，Sigma公司
c)受试动物：SD大鼠，清洁级，雄性，体重约220g
2、SD大鼠药代动力学试验方法：
化合物11、化合物16、对照例44化合物和DHTBZ分别灌胃给药雄性SD大鼠(每组4只动物)，给药剂量为10μmol/kg，给药后0.25,0.5,1,2,3,4,6,8,12h采血约0.2mL至肝素化的试管中，半小时内将样品离心分离血浆至1.5ml EP管中，-20℃保存待检。
3、SD大鼠脑组织分布试验方法
VBZ、化合物21、化合物22、化合物23和对照例45化合物灌胃给药雄性SD大鼠(每组3只动物)，给药剂量为12μmol/kg，给药后0.5h,2h,6h采血约1mL至肝素化的试管中，半小时内将样品离心分离血浆至1.5ml EP管中，-20℃保存待检。大鼠处死后取脑组织，用 生理盐水淋洗脑组织，滤纸吸干，剥离掉脑血管，脑组织称重后-20℃保存。
4、样品分析
样品前处理采用沉淀蛋白的方法，简单描述如下：25微升血浆/50微升脑组织匀浆液用200微升/400微升含内标的乙腈进行沉淀蛋白，高速离心后上清液用水1:1(V/V)稀释后进样分析。
采用LC-MS/MS法测定0.5,2,6h时SD大鼠血浆及脑组织匀浆液(1:4 w/v的生理盐水匀浆)中VBZ、化合物21、化合物22、化合物23和对照例45化合物的浓度，以及它们分别对应的代谢产物DHTBZ、化合物11、化合物12、化合物13和对照例44化合物的浓度，并计算每个时间点脑内和血浆浓度的比值
5、试验结果
5.1药代试验
SD大鼠分别灌胃给药化合物11、化合物16、对照例44化合物和DHTBZ的试验结果见图9，试验结果显示：相对于对照例44化合物和DHTBZ，化合物11有更高的暴露量。
5.2脑组织分布试验
5.2.1脑内浓度：
SD大鼠分别灌胃给药VBZ、化合物21、化合物22、化合物23和对照例45化合物后，原化合物脑组织中药物分布见图10；活性代谢物(DHTBZ、化合物11、化合物12、化合物13和对照例44化合物)脑组织中药物组织分布见图11。
试验结果显示：给药后2h，化合物21及其代谢物化合物11的脑组织浓度均明显高于VBZ及DHTBZ。
5.2.2脑血比
SD大鼠分别灌胃给药VBZ、化合物21、化合物22、化合物23和对照例45化合物后，原化合物脑血比见图12；试验结果显示：在给药后各时间点，化合物21的脑血比是VBZ的3.4～6.8倍，也高于其他化合物的脑血比。
SD大鼠分别灌胃给药VBZ、化合物21、化合物22、化合物23和对照例45化合物后，活性代谢物(DHTBZ、化合物11、化合物12、化合物13和对照例44)脑血比见图13；试验结果显示：在给药后各时间点，化合物21的代谢产物化合物11脑血比是DHTBZ的2.8-5.8倍。
试验例3 化合物21-P3、11-P4SD大鼠药代动力学评价
1.试验材料：
a)受试化合物
11-P4：按照实施例29制备得到
21-P3：按照实施例31制备得到
VBZ：按照试验例1提及的方法制备
DHTBZ：江苏威凯尔医药科技有有限公司，批号67-25-1521-59C
b)溶媒：20％solutol溶液，Solutol批号：BCBQ5646V，Sigma公司
c)受试动物：SD大鼠，清洁级，雄性，体重约220g，24只，随机分组，每组3只(3只灌胃，3只静脉给药)。
2.试验方法：
(1)药液配置：21-P3、11-P4、VBZ及DHTBZ分别精密称定约20mg，用适量溶媒溶解，使其浓度均为1μmol/mL。
(2)生物利用度试验：灌胃给药体积为5mL/kg，给药剂量为5μmol/kg，每个化合物给药3只动物；静脉给药体积为2mL/kg，给药剂量为2μmol/kg，每个化合物给药3只动物；给药前SD大鼠禁食12h，自由饮水，给药后3h统一进食。给药后，分别在对应的时间点采血约1ml至肝素化的试管中，半小时内将样品离心分离血浆至1.5ml EP管中，-20℃保存待检。
3.样品分析
采用LC-MS/MS法测定SD大鼠血浆中化合物VBZ、DHTBZ、21-P3、11-P4的浓度，VBZ和21-P3给药组同时测定分别对应的代谢产物DHTBZ、11-P4的浓度。样品前处理采用以下沉淀蛋白的方法：25微升血浆用200微升/含内标的乙腈进行沉淀蛋白，高速离心后上清液用水1:1(V/V)稀释后进样分析。
4.试验结果：
表31：SD大鼠灌胃给予不同化合物(5μmol/kg)后平均血药浓度(nmol/L)
Figure PCTCN2020108314-appb-000120
BLQ表示在此时间点未取样；NA表示在此时间点数据不可用。
表32：SD大鼠静脉给予不同化合物(2μmol/kg)后平均血药浓度(nmol/L)
Figure PCTCN2020108314-appb-000121
Figure PCTCN2020108314-appb-000122
BLQ表示在此时间点未取样；NA表示在此时间点数据不可用。
表33 SD大鼠静脉给予不同化合物(2μmol/kg)后的药动学参数
Figure PCTCN2020108314-appb-000123
*转化率＝代谢产物AUC last/(原型药物AUC last+代谢产物AUC last)×100％
静脉给药转化率(VBZ)＝508.7/(508.7+2691.7)＝15.9％；
静脉给药转化率(21-P3)＝713.5/(713.5+1952.1)＝26.8％
表34：SD大鼠灌胃给予不同化合物(5μmol/kg)后的药动学参数
Figure PCTCN2020108314-appb-000124
大鼠灌胃给药的生物利用度(BA)＝灌胃AUC last/静脉AUC last×100％
BA(VBZ)＝(2019.1+1110.5)/(2691.7+508.7)/2.5＝39.1％；
BA(21-P3)＝(2541.9+1488.3)/(1952.1+713.5)/2.5＝60.5％；
BA(DHTBZ)＝2381/1544.4/2.5＝61.7％；
BA(11-P4)＝8039/4251.6/2.5＝75.6％。
结果表明：(1)化合物11-P4可显著提高体内暴露量(AUC)，静脉给药的暴露量是DHTBZ的近3倍，灌胃给药的暴露量是DHTBZ的近4倍；
(2)通过静脉给药，化合物11-P4具有更长的半衰期，因此可降低给药频率；
VBZ转化为DHTBZ的转化率为15.9％，化合物21-P3转化为11-P4的转化率为26.8％，相对于VBZ，化合物21-P3转化率更高。由于VBZ和21-P3对VMAT2的结合活性远低于DHTBZ和11-P4，因此，试验结果说明，在等摩尔剂量下，相对于VBZ，化合物21-P3将有更高的利用率，可产生更高的药效。
(3)通过灌胃给药，相对于VBZ和DHTBZ，化合物21-P3和化合物11-P4具有更高的生物利用度
试验例4 化合物21、21-P3的SD大鼠药效学评价
1.目的
本研究采用SD大鼠自主活动模型，以VBZ二甲苯磺酸盐为对照，单次灌胃给予相等摩尔 剂量的化合物21、21-P3，比较大鼠在旷场内的运动距离差异，考察化合21、21-P3与对照药物VBZ二甲苯磺酸盐药效学差异。
2.试验材料
2.1试验动物：SD大鼠，SPF级，雄性32只，周龄5-7周，体重：200-220g，在试验开始前至少预适应1周。动物来源：济南朋悦试验动物技术有限公司；动物合格证号码：SCXK(鲁)20140007
2.2试验药物
化合物21：按照实施例21制备得到
化合物21-P3：按照实施例31制备得到
VBZ二甲苯磺酸盐：江苏威凯尔医药科技有有限公司，批号334-1-1517-15
配制方法：称取适量药物，加少量DMSO(不超过总体积1％)溶解后，加入20％Solutol至所需药物浓度，DMSO终浓度<4％
3.试验分组及给药
试验前一天根据大鼠体重随机分为4组：对照组(NS，溶剂中未添加受试药物)，VBZ二甲苯磺酸盐组，21-P3组，各组均8只动物。大鼠放入检测箱预适应10min，禁食。动物分组及给药信息详见表3。
表35：动物分组与给药
Figure PCTCN2020108314-appb-000125
4.试验方法
试验当天，动物在测试试验室中至少适应1hr。各组大鼠按表3剂量分别单次灌胃给予溶媒或相应药物后放入活动室，使用TopScan监测系统记录并分析大鼠在给药后2hr～3hr内的总运动距离。避免八个活动室内放同一组大鼠，每轮试验至少有一只对照组大鼠，防止相互干扰。在每组试验结束时，将粪便清洗干净以避免无关干扰因素(气味等)对大鼠运动活动的影响。
5.观察指标
使用TopScan监测系统记录并分析大鼠在给药后2hr～3hr内的总运动距离，并计算总运动距离减少率(Reduction Rate，RR)，RR＝(对照组运动距离-给药组运动距离)/对照组运动距离*100％。
6.统计分析
试验数据以均数±标准误(MEAN±SEM)表示，采用PASW stastics 18.0软件单因素方差分析ANOVA比较在每个时间点各组间的差异，所有检验均为双侧检验，P<0.05表示差异具 有统计学显著性意义。
7.试验结果：
与对照组相比(移动距离＝16210±3465mm)，VBZ组可显著降低大鼠自主活动距离(移动距离＝4882±1022mm，P<0.05)；21组自主活动距离降低(移动距离＝11630±2839mm，P>0.05)；21-P3组能显著降低大鼠的自主活动距离(移动距离＝2956±1101mm，P<0.01)，与对照组相比存在显著性差异。VBZ组、21组和21-P3组的总运动减少率分别为69.9％、28.3％和81.8％。相对于VBZ组，21-P3组具有更强的药效。
试验例5 化合物11-P4的SD大鼠药效学评价
试验方法同试验例4，动物分组及给药信息见表36。
表36 动物分组及给药信息
Figure PCTCN2020108314-appb-000126
记录并分析大鼠在给药后0hr～1hr内的总运动距离。试验结果见表37，与对照组相比(移动距离＝14190±2785mm)，VBZ组2.5μmol/kg、5.0μmol/kg及10.0μmol/kg三个剂量组大鼠自主活动距离均降低，其中5.0μmol/kg及10.0μmol/kg组与对照组相比存在显著性差异(移动距离分别为8349±2536mm，P>0.05；6365±2564mm，P<0.05；6742±892.6mm，P<0.05)。11-P4的1.25μmol/kg、2.5μmol/kg、5.0μmol/kg及10.0μmol/kg四个剂量组大鼠的自主活动距离均降低，除1.25μmol/kg组外，其他三个剂量组与对照组相比均存在显著性差异(移动距离分别为9313±1213mm，P>0.05；5959±1615mm，P<0.05；1216±429.9mm，P<0.01；1355±524.1mm，P<0.01)。
表37 各组大鼠总运动距离减少率
Figure PCTCN2020108314-appb-000127
计算不同剂量药物对大鼠自主活动的抑制率，抑制率＝总运动距离减少率(Reduction Rate，RR)，RR＝(对照组运动距离-给药组运动距离)/对照组运动距离*100％，得出VBZ及11-P4的剂量效应曲线，见图14，使用软件Graph Pad Prism 5软件中"log(inhibitor)vs.response-Variable slope"方法计算ED50值。
试验结果显示，等摩尔给药条件下，VBZ组及11-P4的ED 50值分别为5.142μmol/kg及1.883μmol/kg，11-P4效果明显优于VBZ组。
试验例6 五个种属肝微粒体温孵试验
1、试验化合物
化合物11-13、15-16，对照例44化合物：按照上述对应的实施例制备得到。
2、试验过程
温孵：100μL温孵体系包括：0.5mg/mL肝微粒体(人，大鼠，小鼠，猴或犬)，100mM的磷酸钠缓冲液，10mM的氯化镁。在温孵体系中加入化合物11-13、15-16，对照例44化合物，使其终浓度1μM，在37℃预温孵3min后，加入NADPH使其终浓度为1mM，启动反应，温孵0，5min，15min,30min,60min，加入200μL冰乙腈终止反应，存放于-20℃待测。每样品每时间点平行处理2份，并设置不加辅酶的空白对照组、阳性对照组。
3、样品分析及数据处理
乙腈终止后的样品，加入一定量的内标，13000rpm离心10min后，上清液用水1:1(V/V)稀释后，采用LC-MS/MS法测定化合物11-13、15-16，对照例44化合物的浓度。以相对剩余量为纵坐标、时间为横坐标，采用半对数作图法计算各个化合物的消除速率常数k，用公式t 1/2＝0.693/k,各化合物的肝微粒体温孵体系下消除半衰期(t 1/2)结果见表38.
结果表明：化合物11在小鼠和猴的肝微粒体中比DHTBZ具有更长的半衰期，此外化合物12、化合物13也有较好的肝微粒体稳定性。
表38：各化合物在不同种属肝微粒的消除半衰期(t 1/2,min)
种属 化合物11 化合物12 化合物16 化合物13 化合物15 对照例44 DHTBZ
人 161.2 ～ 86.6 154.0 39.4 157.5 ～
大鼠 ～ ～ ～ ～ 67.9 ～ ～
小鼠 23.0 29.9 19.4 78.8 5.1 22.6 13.4
犬 ～ ～ 126.0 ～ 22.6 64.2 115.5
猴 58.7 85.6 14.3 50.6 7.4 26.2 26.3
“～”，数据表明代谢稳定，温孵60min后原化合物剩余率大于80％
试验例7 化合物11-P4与化合物11-P4S晶型A辅料相容性试验
1.试验药物
化合物11-P4：按照实施例29制备
化合物11-P4S晶型A：按照实施例30制备
2.试验方法
将化合物11-P4、化合物11-P4S晶型A分别与辅料以1:20的比例混合，之后在60℃的 条件下敞口放置5天，取样检测化合物11-P4/化合物11-P4S晶型A的纯度。
纯度检测方法：取样品约35mg，精密称定，置25ml量瓶中，加15ml乙腈超声溶解，用水稀释至刻度，摇匀，作为供试品溶液。照高效液相色谱法测定，用十八烷基硅烷键合硅胶为填充剂(Inerstil ODS-3V,250mm×4.6mm，5μm)；以10mmol/L磷酸氢二铵溶液(调节pH值至6.95±0.05)-乙腈(80：20)为流动相A，以乙腈为流动相B，按表39进行梯度洗脱；检测波长为282nm；柱温35℃；流速1.0ml/min。精密量取供试品溶液10μl，注入液相色谱仪，记录色谱图。按峰面积归一化法计算。
表39 流动相梯度
时间(min) 流动相A(％) 流动相％(B)
0 75 25
10 75 25
30 25 75
45 25 75
47 75 25
60 75 25
3.试验结果见表40
表40 辅料相容性试验结果
Figure PCTCN2020108314-appb-000128
结果表明：11-P4与上述辅料混合后经高温放置5天后，纯度明显下降，而11-P4S晶型A纯度无明显变化，说明11-P4S晶型A具有更好的辅料相容性，更便于制剂的开发。
试验例8 11-P4S晶型A/D/E混悬竞争试验
1.试验药物
化合物11-P4S晶型A/D/E：按照实施例30制备
2.试验方法：
在2毫升11-P4S晶型A的IPA和IPAc饱和溶液中，分别加入11-P4S晶型A/D/E各约5mg，室温/50℃悬浮搅拌17小时后测试固体XRPD。
3.试验结果见表41和图15。
表41 混悬竞争结果
Figure PCTCN2020108314-appb-000129
结果显示：所有体系均得到11-P4S晶型A晶型A，表明11-P4S晶型A为室温至50℃下热力学最稳定的晶型。

Claims (13)

  1. 式(I)所示化合物或其立体异构体或其药学上可接受的盐，
    Figure PCTCN2020108314-appb-100001
    其中，
    “---”选自：单键或双键；
    “---”为单键时，R选自OH、H或
    Figure PCTCN2020108314-appb-100002
    “---”为双键时，R为O；
    R 1选自氢、甲基或乙基；
    R 2选自未被取代或者被1、2或3个R 3取代的C 2-10烷基、C 3-6环烷基、C 3-6环烷基-C 1-6烷基、3～6元杂环烷基-C 1-3烷基、C 2-6烯基或C 1-6杂烷基；
    R 3选自F、Cl、Br、OH、SH或NH 2。
  2. 根据权利要求1所述化合物或其立体异构体或其药学上可接受的盐，其中，R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选为未被取代或被2-3个R 3取代的C 2-5烷基；更优选为乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基；最优选为三氟乙基或环丙烷亚甲基。
  3. 根据权利要求1或2任一所述化合物或其立体异构体或其药学上可接受的盐，其中R1为甲基；“---”为单键，R选自OH或
    Figure PCTCN2020108314-appb-100003
    或“---”为双键，R为O。
  4. 根据权利要求1所述化合物或其立体异构体或其药学上可接受的盐，所述的化合物选自：
    Figure PCTCN2020108314-appb-100004
    Figure PCTCN2020108314-appb-100005
    Figure PCTCN2020108314-appb-100006
  5. 根据权利要求1至4任一所述化合物的对甲基苯磺酸盐，优选以下化合物：
    Figure PCTCN2020108314-appb-100007
  6. 结晶形式的下式化合物：
    Figure PCTCN2020108314-appb-100008
  7. 根据权利要求6所述化合物的晶型A，其属于正交晶系P21212空间群，
    Figure PCTCN2020108314-appb-100009
    Figure PCTCN2020108314-appb-100010
    α＝90°,β＝90°,γ＝90°,
    Figure PCTCN2020108314-appb-100011
    Z＝4。
  8. 根据权利要求6所述化合物的晶型A，通过Cu-Ka辐射得到的X射线粉末衍射图谱中包含以下2θ反射角测定的特征峰：6.33±0.2°，10.87±0.2°，18.89±0.2°。
  9. 根据权利要求6所述化合物的晶型A，通过Cu-Ka辐射，具有基本如图2-1所示的X射线粉末衍射图谱。
  10. 一种药物组合物，包括权利要求1～5任一所述化合物或其立体异构体或其药学上可接受的盐，或权利要求6～9任一所述结晶形式化合物，以及药学上可接受的载体。
  11. 根据权利要求1～5任一所述的化合物或其立体异构体或其药学上可接受的盐，或权利要求6～9任一所述结晶形式化合物，或权利要求10所述的药物组合物在制备治疗与VMAT2相关疾病的药物的应用。
  12. 根据权利要求1～5任一所述的化合物或其立体异构体或其药学上可接受的盐，或权利要求6～9任一所述结晶形式化合物，或权利要求10所述的药物组合物在制备治疗运动机能亢进性障碍的药物的应用；优选所述运动机能亢进性障碍包括亨廷顿舞蹈症、迟发性运动障碍、图雷特综合征或抽搐。
  13. 式(II)所述化合物的制备方法，其中包含以下制备步骤：
    Figure PCTCN2020108314-appb-100012
    其中，R 2选自未被取代的C 2-10烷基、C 3-6环烷基-C 1-6烷基，或者被1、2或3个R 3取代的C 2-10烷基；优选为未被取代或被2-3个R 3取代的C 2-5烷基；R 3选自F、Cl、Br、OH或NH 2；
    R 2更优选为乙基、丙基、异丁基、单氟丁基、单氟戊基、三氟乙基、三氟丙基、三氟丁基、三氟戊基、双氟乙基或环丙烷亚甲基，最优选为三氟乙基或环丙烷亚甲基；
    X为离去基团，优选为Cl、Br、I。
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022203984A1 (en) * 2021-03-22 2022-09-29 Neurocrine Biosciences, Inc. Vmat2 inhibitors and methods of use
WO2024064178A1 (en) * 2022-09-21 2024-03-28 Neurocrine Biosciences, Inc. Hexahydro-2h-pyrido[2,1-a]isoquinoline vmat2 inhibitors and methods of use
WO2026050236A1 (en) 2024-08-27 2026-03-05 Neurocrine Biosciences, Inc. Muscarinic receptor agonist in combination with a vesicular monoamine transporter 2 inhibitor, for use in the treatment of a neurological or psychiatric disorder

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12565495B2 (en) * 2019-08-12 2026-03-03 Luye Innomind Pharma Shijiazhuang Co., Ltd. VMAT2 inhibitor and preparation method therefor and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101484190A (zh) * 2006-05-02 2009-07-15 宾夕法尼亚大学理事会 放射性标记的二氢丁苯那嗪衍生物及其作为显像剂的用途
WO2018222549A1 (en) * 2017-05-31 2018-12-06 Five Eleven Pharma Inc. Novel deuterium substituted positron emission tomography (pet) imaging agents and their pharmacological application

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6087376A (en) * 1997-02-05 2000-07-11 University Of Kentucky Research Foundation Use of lobeline compounds in the treatment of central nervous system diseases and pathologies
EP2081929B1 (en) * 2006-11-08 2013-01-09 Neurocrine Biosciences, Inc. Substituted 3-isobutyl-9, 10-dimethoxy-1,3,4,6,7,11b-hexahydro-2h-pyrido[2,1-a]isoquinolin-2-ol compounds and methods relating thereto
US8008500B2 (en) * 2007-06-08 2011-08-30 General Electric Company Intermediates useful for making tetrabenazine compounds
GB2463452A (en) 2008-09-08 2010-03-17 Cambridge Lab Desmethyl derivatives of tetrabenazine and pharmaceutical compositions thereof
CN102120742B (zh) * 2010-01-08 2013-03-13 中国药科大学 一种丁苯那嗪的制备方法
CN102285984B (zh) * 2010-11-25 2012-10-10 江苏威凯尔医药科技有限公司 (2R,3R,11bR)-二氢丁苯那嗪及相关化合物的制备方法
SG11202004165UA (en) 2017-11-08 2020-06-29 Yuhua Li Esters of dihydrotetrabenazine
JP7212958B2 (ja) 2017-12-26 2023-01-26 ▲蘇▼州科睿思制▲葯▼有限公司 バルベナジントシル酸塩の結晶形及びその製造方法並びに用途
US12565495B2 (en) * 2019-08-12 2026-03-03 Luye Innomind Pharma Shijiazhuang Co., Ltd. VMAT2 inhibitor and preparation method therefor and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101484190A (zh) * 2006-05-02 2009-07-15 宾夕法尼亚大学理事会 放射性标记的二氢丁苯那嗪衍生物及其作为显像剂的用途
WO2018222549A1 (en) * 2017-05-31 2018-12-06 Five Eleven Pharma Inc. Novel deuterium substituted positron emission tomography (pet) imaging agents and their pharmacological application

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
CHENG, Y.PRUSOFF, W.H., BIOCHEM. PHARMACOL., vol. 22, 1973, pages 3099 - 3108
DATABASE REGISTRY 6 December 2007 (2007-12-06), ANONYMOUS: "2H-Benzo[a]quinolizin-2-ol, 9-[4-(fluoro-18F)butoxy]-1,3,4,6,7,11b- hexahydro-10-methoxy-3-(2-methylpropyl)-, (2R,3R,11bR)-rel- (CA INDEX NAME)", XP055887848, retrieved from STN Database accession no. 956903-28-9 *
DATABASE REGISTRY ANONYMOUS: "/1 -(C) FILE REGISTRY RN -927182-65-8 REGISTRY ED -Entered STN: 18 Mar 2007 CN -2H-Benzo[a]quinolizin-2-ol, 1,3,4,6,7,11b-hexahydro-10-methoxy-9-(3- methoxypropoxy)-3-(2-methylpropyl)-(CA INDEX NAME)", XP055781055 *
HANK F.KUNG ET AL.: "In vivo imaging of vesicular monoamine transporter 2 in pancreas using an 18F epoxide derivative of tetrabenazine transporter 2 in pancreas using an 18F epoxide derivative of tetrabenazine", NUCLEAR MEDICINE AND BIOLOGY, vol. 35, no. 8, 30 November 2008 (2008-11-30), XP025678986, DOI: 10.1016/j.nucmedbio.2008.08.004 *
HAO XUESHI, JIN QIHUI, VA PORINO, LI CHUN, SHEN WEIJUN, LAFFITTE BRYAN, WU TOM Y.-H.: "Pancreas-Specific Delivery of β-Cell Proliferating Small Molecules", CHEMMEDCHEM, WILEY-VCH, DE, vol. 11, no. 11, 6 June 2016 (2016-06-06), DE, pages 1129 - 1132, XP055780343, ISSN: 1860-7179, DOI: 10.1002/cmdc.201600116 *
See also references of EP4015517A4

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022203984A1 (en) * 2021-03-22 2022-09-29 Neurocrine Biosciences, Inc. Vmat2 inhibitors and methods of use
US11718618B2 (en) 2021-03-22 2023-08-08 Neurocrine Biosciences, Inc. Substituted pyrido[2,1-a]isoquinolines as VMAT2 inhibitors
JP2024511053A (ja) * 2021-03-22 2024-03-12 ニューロクライン バイオサイエンシーズ，インコーポレイテッド Ｖｍａｔ２阻害剤および使用方法
EP4606429A3 (en) * 2021-03-22 2025-12-03 Neurocrine Biosciences, Inc. Vmat2 inhibitors and methods of use
WO2024064178A1 (en) * 2022-09-21 2024-03-28 Neurocrine Biosciences, Inc. Hexahydro-2h-pyrido[2,1-a]isoquinoline vmat2 inhibitors and methods of use
WO2026050236A1 (en) 2024-08-27 2026-03-05 Neurocrine Biosciences, Inc. Muscarinic receptor agonist in combination with a vesicular monoamine transporter 2 inhibitor, for use in the treatment of a neurological or psychiatric disorder

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