CN115707712A - Peptide amide compound, pharmaceutically acceptable salt and stereoisomer thereof, and application thereof - Google Patents
Peptide amide compound, pharmaceutically acceptable salt and stereoisomer thereof, and application thereof Download PDFInfo
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Abstract
The invention relates to peptide amide compounds, pharmaceutically acceptable salts, stereoisomers and application thereof, wherein the cyclic group substituted polyamide compounds have a structure shown in a formula (II)
Description
Technical Field
The invention relates to the technical field of pharmaceutical chemistry, in particular to a peptide amide compound, pharmaceutically acceptable salts and stereoisomers thereof and application thereof.
Background
Opioid receptors can be mainly divided into four receptors of classical receptors mu, delta, kappa and non-classical Nociceptin (NOP), and the receptors of mu, delta and kappa have different subtypes, wherein the kappa opioid receptor agonist has the advantages of weak respiratory depression and low addiction besides the analgesic effect. In addition, the kappa opioid receptor agonist also has anti-inflammatory activity, can be applied to preventing and treating gastrointestinal diseases such as pruritus caused by nephropathy such as cardiovascular diseases and uremia, irritable bowel syndrome, intestinal obstruction and the like, and has better application prospect in developing selective kappa opioid receptor agonists. Kappa opioid receptors have multiple subtypes. In addition to classical kappa 1, there are also the kappa 2 and kappa 3 subtypes, and a distribution in human brain and placental tissues. Kappa opioid receptors are involved in analgesia and are involved in neuroendocrine and immune regulation. Intein-ergenic neurons → release intein (enkephalin) → agonizing opioid receptors → by G protein coupling mechanism → inhibit AC → Ca influx ↓, kappa efflux ↓ → promastic transmitter (substance P etc.) release ↓ → postsynaptic membrane hyperpolarization → prevent conduction, transmission of pain impulse → analgesia.
In primates, opioids have 3 receptors: μ (MOR), kappa (KOR), and Delta (DOR). Opioids (heroin, morphine, codeine) can cause the occurrence of itching. Systemic application of mu receptor agonists induces scratching behavior, whereas kappa receptor agonists and delta receptor agonists inhibit scratching. This phenomenon is not mediated by histamine. In contrast, kappa receptor agonists, like mu receptor antagonists, can inhibit opioid-induced itch by prophylaxis or antagonism. The objectives of the development and application of these agonists include: treating heroin and alcohol dependence, and resisting depression after anesthesia, narcotic overdose and opium drug detoxification.
Physiological opioids (nalorphine, endorphin, dynorphin) are involved in the pathophysiological mechanisms of itching, in particular Atopic Dermatitis (AD), uremic pruritus and hepatic pruritus. The main peptide substances are beta endorphin (MOR agonist), dynorphin a (KOR agonist). They act at central, spinal and peripheral levels. Non-histamine dependent opioid-dependent pruritus appears to arise from cutaneous unmyelinated C fibers. Behavioral experiments explained that KOR and MOR knock-out mice had a significant reduction in stable wild-type scratching in the xerodermic dermatitis model. These results reveal that: MOR and KOR are important for skin homeostasis, epidermal nerve fiber regulation, and itch pathophysiology. In AD patients, there is only downregulation of the kappa receptor system, but not the mu receptor system. Further studies have shown that patients treated with psoralen and uv light have a recovery of the kappa receptor system with a concomitant reduction in the visual analogue score for pruritus. These results suggest that opioids in the epidermis are closely related to the regulation of itching in AD patients. Therefore, there is an urgent need to develop a series of kappa opioid receptor agonists that would be useful in the treatment of the above-mentioned diseases.
Disclosure of Invention
Based on the above, there is a need for peptide amides having kappa opioid receptor agonistic activity, pharmaceutically acceptable salts, stereoisomers and applications thereof.
A compound having a structure represented by formula (II), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:
X 1 and X 2 In, one is CH 2 One is NR, wherein R is
A is
R 1 And R 2 Each independently is: H. c 1-6 Alkyl, - (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Or- (CH) 2 ) n4 -NR b R c ；
n1, n2, n3, n4 are each independently 1,2,3,4 or 5;
Y 1 is a single bond, CR 11 R 12 、O、S、-(CR 11 ＝CR 12 ) -or NH-;
R 11 and R 12 Each independently selected from: H. c 1-4 Alkyl, phenyl, 5-6 membered heteroaryl or halogen; said C is 1-4 Alkyl, phenyl, 5-6 membered heteroaryl optionally further substituted with; halogen, cyano, hydroxy, phenyl, -NR b R c 、-COR d 、-COOR d or-CONR b R c ；
B is selected from: 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl;
R 0 is a substituent on B, m1 is 0, 1,2,3,4, 5 or 6;
R 0 at each occurrence, each group is independently selected from: c 1-4 Alkyl radical, C 1-4 Alkoxy, halogen, cyano, hydroxy, phenyl, 5-6 membered heteroaryl, = O, -NR b R c 、-COR d 、-COOR d 、-CONR b R c or-NHCOR d (ii) a Said C is 1-4 Alkyl radical, C 1-4 Alkoxy, phenyl, 5-6 membered heteroaryl optionally further substituted with: halogen, cyano, hydroxy or phenyl;
R a is C 1-4 An alkyl group;
R b 、R c each independently selected from: H. c 1-4 Alkyl or C 1-4 An alkoxy group;
R d selected from: h or C 1-4 An alkyl group.
The invention provides a pharmaceutical composition, which comprises the compound shown above, or a stereoisomer and a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers and/or excipients.
The invention provides the use of a compound as described above, or a stereoisomer, pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, in the manufacture of a medicament for treating or preventing a disease or condition associated with the kappa opioid receptor in a mammal.
Detailed Description
In order that the invention may be more fully understood, a more complete description of the invention, and a preferred embodiment of the invention, is now provided. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Interpretation of terms
Unless otherwise indicated or contradicted, terms or phrases used herein have the following meanings:
in the present invention, "optionally further substituted" means that it may or may not be substituted, for example: c 1-6 Alkyl is optionally further substituted by R 0 Substituted, represents C 1-6 Alkyl may be substituted by R 0 Substituted or not substituted by R 0 And (4) substitution.
In the present invention, when the number of substituents is not indicated, it means that the substituents are substituted by optionally substitutable number of substituents, such as halogen substituted methyl, which means that the methyl group may be substituted by 1,2 or 3 halogens; when the selected substituent is a substituent group, it means that an optional number of substituents selected from the substituent group may be substituted, for example, R 0 Substituted phenyl, R 0 Selected from: halogen, hydroxy, cyano or NH 2 The term "substituted" means that the phenyl group may be substituted with only one group (e.g., halogen) or may be simultaneously substituted with a plurality of groups (e.g., halogen and amino), and the substitution site is not particularly limited, and only a group capable of stably existing is required to be composed.
In the present invention, when the substitution site is not specified, it means that the optionally substitutable site is substituted.
The term "alkyl" refers to a saturated hydrocarbon containing a primary (normal) carbon atom, or a secondary carbon atom, or a tertiary carbon atom, or a quaternary carbon atom, or a combination thereof. Phrases containing the term, e.g., "C 1 ～C 6 Alkyl "means an alkyl group containing from 1 to 6 carbon atoms, which may, for each occurrence, be independently C 1 Alkyl radical, C 2 Alkyl radical, C 3 Alkyl radical, C 4 Alkyl radical, C 5 Alkyl radical, C 6 An alkyl group. Further, it is preferably C 1-4 An alkyl group; more preferably C 1-3 An alkyl group. Suitable examples of alkyl groups include, but are not limited to: methyl (Me, -CH) 3 ) Ethyl (Et-CH) 2 CH 3 ) 1-propyl (n-Pr, n-propyl, -CH) 2 CH 2 CH 3 ) 2-propyl (i-Pr, i-propyl, -CH (CH) 3 ) 2 ) 1-butyl (n-Bu, n-butyl, -CH) 2 CH 2 CH 2 CH 3 ) 2-methyl-1-propyl (i-Bu, i-butyl, -CH) 2 CH(CH 3 ) 2 ) 2-butyl (s-Bu, s-butyl, -CH (CH) 3 )CH 2 CH 3 ) 2-methyl-2-propyl (t-Bu, t-butyl, -C (CH) 3 ) 3 ) 1-pentyl (n-pentyl, -CH) 2 CH 2 CH 2 CH 2 CH 3 ) 2-pentyl (-CH (CH) 3 )CH 2 CH 2 CH 3 ) 3-pentyl (-CH (CH) 2 CH 3 ) 2 ) 2-methyl-2-butyl (-C (CH) 3 ) 2 CH 2 CH 3 ) 3-methyl-2-butyl (-CH (CH) 3 )CH(CH 3 ) 2 ) 3-methyl-1-butyl (-CH) 2 CH 2 CH(CH 3 ) 2 ) 2-methyl-1-butyl (-CH) 2 CH(CH 3 )CH 2 CH 3 ) 1-hexyl (-CH) 2 CH 2 CH 2 CH 2 CH 2 CH 3 ) 2-hexyl (-CH (CH) 3 )CH 2 CH 2 CH 2 CH 3 ) 3-hexyl (-CH (CH) 2 CH 3 )(CH 2 CH 2 CH 3 ) 2-methyl-2-pentyl (-C (CH)) 3 ) 2 CH 2 CH 2 CH 3 ) 3-methyl-2-pentyl (-CH (CH) 3 )CH(CH 3 )CH 2 CH 3 ) 4-methyl-2-pentyl (-CH (CH) 3 )CH 2 CH(CH 3 ) 2 ) 3-methyl-3-pentyl (-C (CH) 3 )(CH 2 CH 3 ) 2 ) 2-methyl-3-pentyl (-CH (CH) 2 CH 3 )CH(CH 3 ) 2 ) 2, 3-dimethyl-2-butyl (-C (CH)) 3 ) 2 CH(CH 3 ) 2 ) 3, 3-dimethyl-2-butyl (-CH (CH) 3 )C(CH 3 ) 3 And octyl (- (CH) 2 ) 7 CH 3 )。
The term "cycloalkyl" refers to a non-aromatic hydrocarbon containing ring carbon atoms and may be a monocyclic, bicyclic, spirocycloalkyl, or bridged cycloalkyl group. Phrases containing the term, such as, "3-10 membered cycloalkyl" refer to cycloalkyl groups containing 3 to 10 carbon atoms; more preferably 3-8 membered cycloalkyl, more preferably 3,4, 5, 6, 7 or 8 membered cycloalkyl. Suitable examples of cycloalkyl groups include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. In addition, "cycloalkyl" may also contain one or more double bonds (i.e., saturated or partially unsaturated cycloalkyl), representative examples of cycloalkyl containing a double bond include cyclopentenyl, cyclohexenyl, cyclohexadienyl, and cyclobutadienyl, and in some embodiments, cycloalkyl includes: 3-8 membered monocyclic alkyl, 5-6 membered cycloalkyl and 5-6 membered cycloalkyl, and 5-6 membered cycloalkylphenyl.
The term "alkoxy" refers to a group having an-O-alkyl group, i.e., an alkyl group as defined above attached to the parent core structure via an oxygen atom. Phrases containing the term, e.g., "C 1-6 Alkoxy "means that the alkyl moiety contains from 1 to 6 carbon atoms and, at each occurrence, may be independently C 1 Alkoxy radical, C 4 Alkoxy radical, C 5 Alkoxy radical, C 6 An alkoxy group. Suitable examples of alkoxy groups include, but are not limited to: methoxy (-O-CH) 3 or-OMe), ethoxy (-O-CH) 2 CH 3 or-OEt) and tert-butoxy (-O-C (CH) 3 ) 3 or-OtBu).
"alkenyl" means containing a moiety having at least one unsaturation, i.e., a carbon-carbon sp 2 A hydrocarbon of a positive carbon atom, a secondary carbon atom, a tertiary carbon atom or a ring carbon atom of a double bond. Phrases containing the term, e.g., "C 2～4 The radical "means an alkenyl radical having from 2 to 4 carbon atoms which, at each occurrence, may be, independently of one another, C 2 Alkenyl radical, C 3 Alkenyl radical, C 4 An alkenyl group. Suitable examples include, but are not limited to: vinyl (-CH = CH) 2 ) Allyl (-CH) 2 CH＝CH 2 ) Cyclopentenyl (-C) 5 H 7 ) And 5-hexenyl (-CH) 2 CH 2 CH 2 CH 2 CH＝CH 2 )。
"aryl" refers to an aromatic hydrocarbon group derived by removing one hydrogen atom from an aromatic ring compound, and may be a monocyclic aromatic group, or a fused ring aromatic group, or a polycyclic aromatic group, at least one of which is an aromatic ring system for polycyclic ring species. For example, "C 5 ～C 10 Aryl "means an aryl group containing from 5 to 10 carbon atoms, which may, for each occurrence, be independently C 5 Aryl radical, C 6 Aryl radical, C 10 And (4) an aryl group. Suitable examples include, but are not limited to: benzene and naphthalene.
"heteroaryl" means that on the basis of an aryl at least one carbon atom is replaced by a non-carbon atom which may be a N atom, an O atom, an S atom, etc. For example, "C 5 ～C 10 Heteroaryl "means a heteroaryl group containing from 5 to 10 carbon atoms, which may be, independently for each occurrence, C 5 Heteroaryl group, C 6 Heteroaryl, C 7 Heteroaryl or C 8 A heteroaryl group. Suitable examples include, but are not limited to: furan, benzofuran, thiophene, benzothiophene, pyrrole, pyrazole, triazole (e.g., 1,2, 3-triazole, 1,2, 4-triazole, 1,2, 5-triazole, 1,3, 4-triazole), imidazole, oxazole, oxadiazole (e.g., 1,2, 3-oxadiazole, 1,2, 4-oxadiazole, 1,2, 5-oxadiazole, 1,3, 4-oxadiazole), thiazole, tetrazole, indole, carbazole, pyrroloimidazole, pyrrolopyrrole, thienopyrrole, thienothiophene, furothiophene, furofuran, thienofuran, benzisoxazole, benzisothiazole, benzimidazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine, quinoline, isoquinoline, phthalazine, quinoxaline, phenanthridine, primary pyridine, quinazoline, and quinazolinone. It is understood that, in the present invention, heteroaryl includes monocyclic heteroaryl, bicyclic heteroaryl (i.e., fused bicyclic heteroaryl), and the group forming fused bicyclic heteroaryl is preferably 5 or 6 membered monocyclic heteroaryl, including but not limited to:
wherein
The two carbon atoms attached represented are adjacent pairs of carbon atoms shared when fused to other rings.
"Heterocyclyl" means that at least one carbon atom is replaced with a non-carbon atom, which may be a N atom, an O atom, an S atom, etc., and may be a saturated ring or a partially unsaturated ring, in addition to a cycloalkyl group. May contain on the heterocycloalkyl ringKetone groups, for example: pyrrolidone group and piperidone group may be omitted. The phrase comprising the term, for example, "3-10 membered saturated or partially unsaturated" means a heterocyclic group containing 4 to 9 carbon atoms, which at each occurrence, independently of each other, may be C 4 Heteroalkyl group, C 6 Heteroalkyl group, C 7 Heteroalkyl group, C 8 Heteroalkyl radicals or C 9 A heteroalkyl group. Suitable examples include, but are not limited to: aziridinyl, oxiranyl, azetidinyl, oxetanyl, oxazolidinyl, 1, 3-dioxolanyl, dioxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl-1, 1-dioxide, tetrahydropyranyl, 1, 3-oxazinoalkyl, hexahydropyrimidyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, 1, 3-oxazepanyl, 1, 2-dihydroazetidinyl, 2, 5-dihydro-1H-pyrrolyl, 2, 3-dihydro-1H-pyrrolyl, 1,2,3, 4-tetrahydropyridinyl, 1,2,3, 6-tetrahydropyridinyl, 2-pyrrolidinonyl or 2-piperidinonyl, tetrahydroisoquinolinyl, indolinyl.
The "heterocyclic group" in the present invention includes, but is not limited to, monocyclic heterocyclic groups and bicyclic heterocyclic groups. In some embodiments, monocyclic heterocyclyl includes 5-6 membered monocyclic heterocyclyl; in some embodiments, bicyclic heterocyclyl groups include 5-6 membered heterocyclo 5-6 membered heterocyclyl, 5-6 membered cycloalkylo 5-6 membered heterocyclyl, 5-6 membered heterocyclo phenyl, 5-6 membered heterocyclo 5-6 membered heteroaryl;
wherein, 5-6 membered cycloalkyl and 5-6 membered heterocyclyl include, but are not limited to:
5-6 membered heterocyclo and 5-6 membered heterocyclo include, but are not limited to:
the 5-6 membered heterocycloocene radical includes, but is not limited to:
"amino" refers to a derivative of ammonia having the formula-N (X) 2 Wherein each "X" is independently H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, or the like. Non-limiting types of amino groups include-NH 2 -N (alkyl) 2 NH (alkyl), -N (cycloalkyl) 2 NH (cycloalkyl), -N (heterocyclyl) 2 NH (heterocyclyl), -N (aryl) 2 NH (aryl), -N (alkyl) (heterocyclyl), -N (cycloalkyl) (heterocyclyl), -N (aryl) (heteroaryl), -N (alkyl) (heteroaryl) and the like.
"halogen" or "halo" refers to F, cl, br, or I.
Detailed explanation
One embodiment of the present invention provides a compound having a structure represented by formula (II):
X 1 and X 2 In, one is CH 2 One is NR, wherein R is
A is
R 1 And R 2 Each independently is: H. c 1-6 Alkyl, - (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Or- (CH) 2 ) n4 -NR b R c ；
n1, n2, n3, n4 are each independently 1,2,3,4 or 5;
Y 1 is a single bond, CR 11 R 12 、O、S、-(CR 11 ＝CR 12 ) -or NH-;
R 11 and R 12 Each independently selected from: H. c 1-4 Alkyl, phenyl, 5-6 membered heteroaryl or halogen; said C is 1-4 Alkyl, phenyl, 5-6 membered heteroaryl optionally further substituted with; halogen, cyano, hydroxy, phenyl, -NR b R c 、-COR d 、-COOR d or-CONR b R c ；
B is selected from: 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl;
R 0 is a substituent on B, m1 is 0, 1,2,3,4, 5 or 6;
R 0 at each occurrence, each group is independently selected from: c 1-4 Alkyl radical, C 1-4 Alkoxy, halogen, cyano, hydroxy, phenyl, 5-6 membered heteroaryl, = O, -NR b R c 、-COR d 、-COOR d 、-CONR b R c or-NHCOR d (ii) a Said C is 1-4 Alkyl radical, C 1-4 Alkoxy, phenyl, 5-6 membered heteroaryl optionally further substituted with: halogen, cyano, hydroxy or phenyl;
R a is C 1-4 An alkyl group;
R b 、R c each independently selected from: H. c 1-4 Alkyl or C 1-4 An alkoxy group;
R d selected from: h or C 1-4 An alkyl group.
In some embodiments, the compound of formula (II) is selected from compounds of any of the following formulae:
in some embodiments, R 1 And R 2 Each independently is: H. c 1-4 Alkyl, - (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Or- (CH) 2 ) n4 -NR b R c ；
In some embodiments, R 1 And R 2 Are all H. In some embodiments, R 1 And R 2 Are all methyl.
In some embodiments, R 1 And R 2 One of them is H and one is- (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Or- (CH) 2 ) n4 -NR b R c 。
In some embodiments, n1 is 1 or 2; in some embodiments, n2 is 2; in some embodiments, n3 is 1 or 2. In some embodiments, n1 is 2, n2 is 2, and n3 is 1,2,3, or 4.
In some embodiments, -NR 1 R 2 Selected from: -NH 2 、-NH-(CH 2 ) 2 -O-(CH 2 ) 2 -O-R a 、-NH(CH 2 ) 3 -NR b R c or-NH (CH) 2 ) 4 -NR b R c ；R a Is C 1-3 An alkyl group; r b 、R c Each independently selected from: h or C 1-3 An alkyl group.
In some embodiments, - (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Is- (CH) 2 ) 2 -O-(CH 2 ) 2 -O-CH 3 。
In some embodiments, n4 is 3. In some embodiments, - (CH) 2 ) n4 -NR b R c Is- (CH) 2 ) 3 -NH 2 。
In some embodiments, B is selected from: 3-8 membered monocycloalkyl, 5-6 membered cycloalkylacenyl, 5-6 membered heteromonocyclic group, 5-6 membered heterocyclo-5-6 membered heterocyclic group, 5-6 membered cycloalkylaceno 5-6 membered heterocyclic group, 5-6 membered heterocyclonophenyl, 5-6 membered heterocyclo 5-6 membered heteroaryl, 6-10 membered aryl, or 5-10 membered heteroaryl.
In some embodiments, B is selected from any of the following:
in some embodiments, R 0 At each occurrence, each group is independently selected from: c 1-4 Alkyl, halogen, -OCF 3 、-NH 2 、-COOR d or-CONR b R c 。
In some embodiments, the compound is selected from the group consisting of compounds represented by any one of the following formulas:
B 1 selected from: 3-8 membered monocycloalkyl, 5-6 membered cycloalkylacenyl, 5-6 membered heteromonocyclic group, 5-6 membered heterocyclo-5-6 membered heterocyclic group, 5-6 membered cycloalkylaceno 5-6 membered heterocyclic group, 5-6 membered heterocyclo-phenyl, 6-10 membered aryl, or 5-10 membered heteroaryl;
B 2 selected from the group consisting of: 6-10 membered aryl or 5-10 membered heteroaryl;
B 3 selected from the group consisting of: 6-10 membered aryl or 5-10 membered heteroaryl;
B 4 selected from: 6-10 membered aryl or 5-10 membered heteroaryl.
In some embodiments, B 3 Selected from the group consisting of: phenyl or 5-6 membered heteroaryl. In some embodiments, B 4 Selected from the group consisting of: phenyl or 5-6 membered heteroaryl.
In some embodiments, the 3-8 membered monocycloalkyl group is selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, or cyclooctyl.
In some embodiments, the 5-6 membered heteromonocyclic group is selected from: aziridinyl, oxiranyl, azetidinyl, oxetanyl, oxazolidinyl, 1, 3-dioxolanyl, dioxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl-1, 1-dioxide, tetrahydropyranyl, 1, 3-oxazinoalkyl, hexahydropyrimidyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, 1, 3-oxazepanyl, 1, 2-dihydroazetidinyl, 2, 5-dihydro-1H-pyrrolyl, 2, 3-dihydro-1H-pyrrolyl, 1,2,3, 4-tetrahydropyridinyl, 1,2,3, 6-tetrahydropyridinyl, 2-pyrrolidinonyl or 2-piperidinonyl;
in some embodiments, the 5-6 membered heterocyclyl and the 5-6 membered heterocyclyl is selected from any one of the following:
in some embodiments, the 5-6 membered cycloalkyl and 5-6 membered heterocyclyl is selected from any one of the following:
in some embodiments, the 5-6 membered cycloalkylacenyl is selected from: a cyclopentylphenyl group, or a cyclohexylphenylphenyl group;
in some embodiments, the 5-6 membered heterocyclo-phenyl is selected from any one of:
in some embodiments, the 6-10 membered aryl is selected from: benzene or naphthalene;
in some embodiments, the 5-10 membered heteroaryl is selected from: thiophene, furan, thiazole, isothiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2, 3-triazole, 1,2, 4-triazole, 1,2, 5-triazole, 1,3, 4-triazole, tetrazole, isoxazole, oxadiazole, 1,2, 3-oxadiazole, 1,2, 4-oxadiazole, 1,2, 5-oxadiazole, 1,3, 4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine, pyrazine or any of the following groups:
in some embodiments of the present invention, the,
selected from any one of the following groups:
z is NH, O, S, CR h R j Or C = O; w is CR h Or N; v is CR h Or N; and two adjacent Z are not hetero atoms at the same time;
R h and R j Each independently selected from: H. c 1-4 Alkyl radical, C 1-4 Alkoxy, halogen, cyano, hydroxy, phenyl, 5-6 membered heteroaryl, -NR b R c 、-COR d 、-COOR d 、-CONR b R c or-NHCOR d (ii) a Said C is 1-4 Alkyl radical, C 1-4 Alkoxy, phenyl, 5-6 membered heteroaryl optionally further substituted with: halogen, cyano, hydroxy, phenyl, -NR b R c or-COOR d 。
In some embodiments, in (A-1), Z is all-CR h R j (ii) a In some embodiments, one of Z in (A-1) is-NH and the others are-CR h R j0 (ii) a In some embodiments, one Z in (A-1) is-NH, one is CO, and the others are-CR h R j0 (ii) a In some embodiments, in (A-2), Z is all-CR h R j (ii) a In some embodiments, one of Z in (A-2) is-NH and the others are-CR h R j (ii) a In some embodiments, one Z in (A-2) is-NH, one is CO, and the others are-CR h R j (ii) a In some embodiments, in (A-3), Z is all-CR h R j (ii) a In some embodiments, 1,2 or 3 of Z in (A-3) are-NH and the remainder are-CR h R j (ii) a In some embodiments, in (A-3), Z is all-CR h R j (ii) a In some embodiments, 1 of (A-3) Z is-NH, one is CO, and the others areis-CR h R j (ii) a In some embodiments, (A-4) wherein Z is all-CR h R j (ii) a In some embodiments, 1,2 or 3 of (A-4) Z are-NH and the remainder are-CR h R j (ii) a In some embodiments, in (A-4), 1Z is NH, one Z is CO, and the remainder are-CR h R j . In some embodiments, (A-5) wherein Z is-CR h R j W is CR h (ii) a In some embodiments, (A-5) wherein Z is O and W are both CR h (ii) a In some embodiments, (A-5) wherein Z is NH and W is CR h (ii) a In some embodiments, (A-6) W is CR h (ii) a In some embodiments, 1,2 or 3 of (A-6) W are N, the remainder being CR h (ii) a In some embodiments, in (A-7), Z is NH and W is CR h (ii) a In some embodiments, (A-7) wherein Z is O and W is CR h (ii) a In some embodiments, in (A-7), one Z is NH, one Z is O, and W are both CR h (ii) a In some embodiments, in (A-8), Z is NH and W is CR h (ii) a In some embodiments, (A-8) wherein Z is O and W is CR h (ii) a In some embodiments, (A-9) wherein Z is all-CR h R j W is CR h (ii) a In some embodiments, (A-10) wherein V is CR h One Z is N, one Z is O and the other Z are-CR h R j (ii) a In some embodiments, (A-10) wherein V is CR h One Z is N and the others are-CR h R j (ii) a In some embodiments, the five-and five-membered ring of (a-10) is a 5-membered nitrogen-containing heterocyclocyclopentane; in some embodiments, the five-and five-membered ring of (a-10) is a 5-membered nitrogen-containing heterocycle and a 5-membered nitrogen-containing heterocycle; in some embodiments, the five-and five-membered ring of (a-10) is a 5-membered nitrogen-containing heterocycle and a 5-membered oxygen-containing heterocycle; in some embodiments, (A-11) wherein V are both CR h One Z is N and the others are-CR h R j (ii) a In some embodiments, (a-11) results in a five-and six-membered ring being a 5-membered nitrogen-containing heterocycle and a 6-membered nitrogen-containing heterocycle; in some embodiments, (A-12) wherein V are both CR h Z is all-CR h R j (ii) a In some embodiments, (A-12) wherein V are both CR h Has 1 portion,2.3 or 4 of Z are N and the remainder are-CR h R j (ii) a In some embodiments, the six-and six-membered ring of (a-12) is a 6-membered nitrogen-containing heterocyclocyclocyclohexane; in some embodiments, the six-and six-membered ring of (a-12) is a 6-membered nitrogen-containing heterocyclic and 6-membered nitrogen-containing heterocycle; in some embodiments, (A-13) wherein Z is all-CR h R j W is CR h (ii) a In some embodiments, 1 or 2 of (A-13) Z are NH and the remainder are-CR h R j W is CR h (ii) a In some embodiments, 1 or 2 of (A-13) Z are O, and the remainder are-CR h R j W is CR h (ii) a In some embodiments, in (A-14), Z is all-CR h R j W is CR h (ii) a In some embodiments, 1 or 2 of (A-14) Z are NH and the remainder are-CR h R j W is CR h (ii) a In some embodiments, 1 or 2 of (A-14) Z are O, and the remainder are-CR h R j W is CR h (ii) a In some embodiments, W in (A-15) is CR h (ii) a In some embodiments, 1,2 or 3 of (A-15) W are N, the remainder being CR h (ii) a In some embodiments, 1,2 or 3 of (A-16) W are N, the remainder being CR h 。
In some embodiments, B is selected from any of the following structures:
the above groups are optionally further substituted by R 0 Substitution; in some embodiments, R 0 At each occurrence, each group is independently selected from: c 1-4 Alkyl, halogen, -OCF 3 、-NH 2 、-COOR d or-CONR b R c 。
In some embodiments of the present invention, the,
selected from any of the following structures:
in some embodiments, R 0 At each occurrence, is independently selected from: c 1-4 Alkyl, halogen, -OCF 3 、-NH 2 、-COOR d 、-CONR b R c or-NHCOR d . In some embodiments, R 0 Selected from the group consisting of: F. cl, -NH 2 、-OCF 3 、-COOCH 3 -COOH or-NHCOCH 3 。
In some embodiments, R a Is methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, or 2-methyl-2-propyl;
in some embodiments, R b 、R c Each independently selected from: H. methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, 2-methyl-2-propyl, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, or 2-methyl-2-propoxy.
In some embodiments, R d Selected from: H. methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2-methyl-1-propyl, 2-butyl, or 2-methyl-2-propyl.
In some embodiments, the compound is selected from any one of:
another embodiment of the invention provides a use of a compound as described above, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating or preventing a disease associated with a kappa opioid receptor.
The invention provides a pharmaceutical composition, which comprises the compound shown as the above, or a stereoisomer and a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers and/or excipients.
In another aspect, the present invention provides the use of a compound as described above, or a stereoisomer, pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, for the manufacture of a medicament for the treatment or prevention of a kappa opioid receptor associated disease or condition in a mammal.
Preferably, wherein the kappa opioid receptor-related disorder is selected from the group consisting of pain, inflammation, itch, edema, hyponatremia, hypokalemia, ileus, cough and glaucoma.
Further, preferred pain is selected from neuropathic pain, trunk pain, visceral pain, skin pain, arthritis pain, kidney stone pain, uterine cramps, dysmenorrhea, endometriosis, dyspepsia, post-surgical pain, post-medical treatment pain, ocular pain, otitis pain, breakthrough cancer pain and pain associated with GI disorders.
The kappa opioid receptor (KOR receptor) -associated pruritus to which the present invention relates may be any pruritus disease and condition, such as ocular pruritus, pruritus associated with end-stage renal disease (where many patients receive renal dialysis) and other forms of cholestasis, skin conditions, insect bite pruritus and drug induced pruritus.
Wherein the ocular pruritus such as conjunctivitis-associated ocular pruritus; other forms of cholestasis include primary biliary cirrhosis, intrahepatic cholestasis of pregnancy, chronic cholesterol liver disease, uremia, malignant cholestasis, jaundice; skin conditions such as eczema (dermatitis), dermatophytosis, polycythemia, lichen planus, chronic lichen simplex, pediculosis, thyrotoxicosis, tinea pedis, urticaria, scabies, vaginitis, and anal pruritus associated with hemorrhoid; among them, eczema (dermatitis) includes atopic dermatitis or contact dermatitis.
The present invention provides a method of treating a disease or condition associated with a kappa opioid receptor comprising the steps of: administering the above compound, or a stereoisomer, a pharmaceutically acceptable salt, or a pharmaceutical composition thereof, to a subject to be administered.
The present invention will be described below with reference to specific examples.
Synthesis of intermediate 1-1
1-1, the synthesis procedure is as follows: weighing 10g of 2-Cl-resin, placing in a polypeptide synthesis tube, adding 50mL of dichloromethane, stirring with nitrogen, adding SM-1 (Fmoc-D-Lys (Boc) -OH,4.68g,10 mmol), DIEA (2.58g, 20mmol), stirring with nitrogen for 1h, adding methanol (5 mL), stirring with nitrogen for 30min, pumping, washing with DMF (50 mL multiplied by 3) for 3 times, pumping, and directly using in the next step; to the polypeptide synthesis tube was added 50mL piperidine/DMF (v: v =1 4), nitrogen was bubbled for 30min, the liquid was drained, DMF (50 mL × 3) was washed 3 times, drained and used directly in the next step; adding 50mLDMF into a polypeptide synthesis tube, stirring nitrogen, adding Fmoc-D-Leu-OH (7.06g, 20mmol), HATU (7.6g, 20mmol), DIEA (2.58g, 20mmol), stirring nitrogen for 1h, draining the reaction solution, washing with DMF (50 mL multiplied by 3) for 3 times, draining, and directly using in the next step; to the polypeptide synthesis tube was added 50mL piperidine/DMF (v: v =1 4), nitrogen was bubbled for 30min, the liquid was drained, DMF (50 mL × 3) was washed 3 times, drained and used directly in the next step; 50mL of DMF was added to the polypeptide synthesis tube, nitrogen purged, fmoc-D-Phe-OH (7.74g, 20mmol), HATU (7.6g, 20mmol), DIEA (2.58g, 20mmol), nitrogen purged for 1h, the reaction solution was drained, and DMF (50 mL. Times.3) was washed 3 times, drained and used directly in the next step; to the polypeptide synthesis tube was added 50mL piperidine/DMF (v: v =1 4), nitrogen was bubbled for 30min, the liquid was drained, DMF (50 mL × 3) was washed 3 times, drained and used directly in the next step; 50mL of DMF was added to the polypeptide synthesis tube, nitrogen gas was bubbled, boc-D-Phe-OH (5.38g, 20mmol), HATU (7.6g, 20mmol), DIEA (2.58g, 20mmol), nitrogen gas was bubbled for 1 hour, and the reaction solution was drained, DMF (50 mL. Times.3) was washed 3 times, drained, added MeOH (50 mL. Times.3) washed 3 times, drained, added DCM (50 mL. Times.3) washed 3 times, drained and used in the next step as is; to the polypeptide synthesis tube was added 50mL hexafluoroisopropanol/DCM (v: v =1, 4), nitrogen was bubbled for 30min, the liquid was aspirated off, repeated 3 times, the resulting liquids were combined and spun dry to give 1-1 (7.01 g, yield: 93.0%) of a white solid, [ M + H ]] + :754.4。
Synthesis of intermediate 1-2
1-2, adopting solid phase synthesis, wherein the synthesis operation is as follows: weighing 1g of 2-Cl-resin, placing the weighed 2-Cl-resin in a polypeptide synthesis tube, adding 5mL of dichloromethane, agitating nitrogen, adding Fmoc-D-Lys (Boc) -OH (0.47g and 1mmol), DIEA (0.26g and 2mmol), agitating nitrogen for 1h, adding methanol (1 mL), agitating nitrogen for 30min, draining, washing with DMF (5 mL multiplied by 3) for 3 times, draining, and directly using in the next step; to the polypeptide synthesis tube was added 5mL piperidine/DMF (v: v =1 4), nitrogen sparged for 30min, liquid was aspirated, DMF (5 mL × 3) washed 3 times, aspirated and used directly in the next step; adding 5mL DMF into a polypeptide synthesis tube, agitating nitrogen, adding Fmoc-D-Leu-OH (0.71g, 2mmol), HATU (0.76g, 2mmol), DIEA (0.26g, 2mmol), agitating nitrogen for 1h, draining the reaction solution, washing 3 times with DMF (5 mL multiplied by 3), draining, and directly using in the next step; to the polypeptide synthesis tube was added 5mL piperidine/DMF (v: v =1 4), nitrogen sparged for 30min, liquid was aspirated, DMF (5 mL × 3) washed 3 times, aspirated and used directly in the next step; adding 5mL of DMF into a polypeptide synthesis tube, bubbling nitrogen, adding Fmoc-D-Phe-OH (0.77g, 2mmol), HATU (0.76g, 2mmol), DIEA (0.26g, 2mmol), bubbling nitrogen for 1h, draining the reaction solution, washing 3 times with DMF (5 mL multiplied by 3), draining, and directly using in the next step; to the polypeptide synthesis tube was added 5mL piperidine/DMF (v: v =1 4), nitrogen sparged for 30min, liquid was aspirated, DMF (5 mL × 3) washed 3 times, aspirated and used directly in the next step; 5mL of DMF was added to a polypeptide synthesis tube, nitrogen gas was bubbled, and (R) -2- ((tert-butoxycarbonyl) (2-phenylpropyl) amino) acetic acid (0.58g, 2mmol), HATU (0.76g, 2mmol), DIEA (0.26g, 2mmol), nitrogen gas was bubbled for 1 hour, the reaction solution was drained, DMF (5 mL. Times.3) was washed 3 times, and MeOH (5 mL. Times.3) was added, washed 3 times, and drained,DCM (5 mL. Times.3) was added and the mixture was washed 3 times and taken in the next step with suction; to the polypeptide synthesis tube was added 5mL hexafluoroisopropanol/DCM (v: v =1 4), nitrogen was bubbled for 30min, the liquid was aspirated off, repeated 3 times, the resulting liquids were combined and spun dry to give 1-2 as a white solid (0.61 g, yield: 78.0%), [ M + H] + :782.4。
Example 1: synthesis of (R) -N- ((R) -6-amino-1- (2- (2- (3, 4-dichlorophenyl) acetyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (176)
(1) Synthesis of tert-butyl 2- (2- (3, 4-dichlorophenyl) acetyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (176-2)
176-1 (188.6mg, 0.92mmol), 1 (250mg, 1.105mmol), HOBT (149mg, 1.105mmol) and EDC. HCl (264.7mg, 1.38mmol) were dissolved in 5mL of dichloromethane, stirred at room temperature for 5 minutes, triethylamine (93mg, 0.92mmol) was added, and the reaction was stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate (5 mL), the organic phase was separated, the aqueous phase was extracted with dichloromethane (5 mL. Times.3), the organic phases were combined, washed with 1N dilute hydrochloric acid (5 mL), and concentrated to give crude 176-2 (312 mg, yield: 81.9%), ESI-MS (m/z): 413.1[ M ] +H] + 。
(2) Synthesis of 2- (3, 4-dichlorophenyl) -1- (2, 7-diazaspiro [3.5] nonan-2-yl) ethanone hydrochloride (176-3)
The crude 176-2 (312mg, 0.76mmol) was dissolved in 5mL of ethyl acetate, and then 2.5mL of dioxane hydrochloride was added, and the reaction was stirred at room temperature for 2 hours and filtered to obtain 176-3 (221 mg, yield: 83.5%) as a white solid, ESI-MS (m/z): 313.1[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (2- (2- (3, 4-dichlorophenyl) acetyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylalanyl) -4-methylpentanamide (176-4)
After 1-1 (300mg, 0.398mmol) and HATU (90.8mg, 0.239mmol) were dissolved in THF (5 mL), DIEA (102.8mg, 0.797mmol) was added thereto, and the reaction mixture was stirred at room temperature for 0.5 hour, then 176-3 (83.4mg.0.239mmol) was added thereto, and the reaction mixture was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (5 mL), the organic phase separated and the aqueous phase diluted with V DCM ：V MeOH Extraction of =10 (5 mL × 3), merging of organic phases, washing with 1N diluted hydrochloric acid (5 mL), concentration, and column chromatography (V) DCM :V MeOH = 20): 50.6%), ESI-MS (m/z): 1048.5[ M ] of +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2- (2- (3, 4-dichlorophenyl) acetyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (176)
After the above-mentioned product 176-4 (211mg, 0.20mmol) was dissolved in 5mL of ethyl acetate, 2.5mL of dioxane hydrochloride solution was added, and the reaction was stirred at room temperature for 2 hours, followed by suction filtration to obtain a brown solid 176 (121 mg, yield: 65.3%), ESI-MS (m/z): 848.4[ 2 ] M + H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.48-7.21(m,13H),4.91-4.87(m,1H),4.75-4.72(m,1H),4.44-4.38(m,1H),4.15-4.11(m,1H),3.79-3.75(m,3H),3.56-3.51(m,4H),3.26-3.23(m,2H),2.99-2.81(m,7H),1.89-1.4(m,13H),0.99(dd,6H)。
Example 2: synthesis of (R) -N- ((R) -6-amino-1- (2- (benzofuran-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (167)
(1) Synthesis of tert-butyl 2- (benzofuran-6-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (167-2)
Mixing 167-1 (149mg, 0.92mmol), 1 (250mg, 1.105mmol), HOBT (149mg, 1.105mmol), EDC & HCl (264.7 mg,1.3 mmol)8 mmol) was dissolved in 5mL of dichloromethane, stirred at room temperature for 5 minutes, added with triethylamine (93mg, 0.92mmol), and stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate (5 mL), the organic phase was separated, the aqueous phase was extracted with dichloromethane (5 mL. Times.3), the organic phases were combined, washed with 1N dilute hydrochloric acid (5 mL), and concentrated to give crude 167-2 (317 mg, yield: 93.2%), ESI-MS (m/z): 371[ M ] +H] + 。
(2) Synthesis of benzofuran-6-yl (2, 7-diazaspiro [3.5] nonan-2-yl) methanone hydrochloride (167-3)
The crude 167-2 (317mg, 0.86mmol) was dissolved in 5mL of ethyl acetate, and then 2.5mL of dioxane hydrochloride solution was added, stirred at room temperature for 2 hours, and filtered to obtain 167-3 (241 mg, yield: 83.6%) as a white solid, ESI-MS (m/z): 271[ 2 ], [ M ] +H] + 。
(3) (R) -N- ((R) -6-tert-Butoxycarbonylamino-1- (2- (benzofuran-5-carbonyl) -2, 7-diazaspiro [ 3.5)]Synthesis of nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-t-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (167-4) 1-1 (300mg, 0.398mmol), HATU (90.8mg, 0.239mmol) were dissolved in THF (5 mL), DIEA (102.8mg, 0.797mmol) was added, and after stirring at room temperature for 0.5 hour, 167-3 (73mg, 0.239mmol) was added, and the reaction was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (5 mL), the organic phase separated and the aqueous phase diluted with V DCM ：V MeOH Extraction of =10 (5 mL × 3), merging of organic phases, washing with 1N diluted hydrochloric acid (5 mL), concentration, and column chromatography (V) DCM :V MeOH = 20): 45.2%), ESI-MS (m/z): 1006.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2- (benzofuran-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (167)
After the above-mentioned product 167-4 (181mg, 0.18mmol) was dissolved in 5mL of ethyl acetate, 2.5mL of dioxane hydrochloride was added, and the reaction mixture was stirred at room temperature for 2 hours and suction-filtered to obtain 167 (101 mg, yield: 63.9%) as a brown solid, ESI-MS (m/z):806.5[M+H] + 。
1 HNMR(400MHz,CD 3 OD),δ8.01-7.98(m,1H),7.89-7.86(m,1H),7.68-7.66(m,2H),7.42-7.21(m,10H),7.01-6.91(m,1H),4.91-4.87(m,1H),4.75-4.72(m,1H),4.44-4.38(m,1H),3.99-3.91(m,1H),3.81-3.62(m,4H),3.26-3.23(m,2H),2.99-2.81(m,8H),1.89-1.4(m,13H),0.99(dd,6H).
example 3:
synthesis of (R) -N- ((R) -6-amino-1- (2- (imidazo [1,2-a ] pyridine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (168)
(1) Synthesis of tert-butyl 2- (imidazo [1,2-a ] pyridine-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (168-2)
After dissolving 168-1 (0.32g, 1.95mmol), HATU (0.80g, 2.12mmol) and DIEA (0.38g, 3.72mmol) in DMF (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 1 (0.40g, 1.77mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC monitored the completion of the reaction, the reaction mixture was diluted with water, extracted with DCM (10 mL. Times.3), the organic phases were combined and then 1N Na was added 2 CO 3 Washed once (10 mL) and once with saturated brine (10 mL), dried the organic phase and spin-dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =100:1 to 30:1 to give 168-2 as a brown solid (0.61 g, yield: 93.1%), ESI-MS (m/z): 371.1[ M ] +H] + 。
(2) Synthesis of imidazo [1,2-a ] pyridin-2-yl (2, 7-diazaspiro [3.5] nonan-2-yl) methanone hydrochloride (168-3)
168-2 (0.61g, 1.65mmol) was dissolved in ethyl acetate (12 mL), and dioxane hydrochloride (4M, 8mL) was added to the solution, after which the reaction mixture was stirred at room temperature for 6 hours. And (5) monitoring by TLC that most of raw materials are completely consumed, stopping the reaction, and directly spin-drying the reaction solution to obtain the product. TargetCompound 168-3 was a white solid (0.55 g, yield: 97.3%), ESI-MS (m/z): 271.0[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (2- (benzofuran-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (168-5)
After 1-1 (250mg, 0.33mmol), HATU (152mg, 0.40mmol) and DIEA (150mg, 1.16mmol) were dissolved in tetrahydrofuran (8 mL), the reaction mixture was stirred at room temperature for 30 minutes, 168-3 (123mg, 0.36mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC monitored for completion of the reaction, and then the reaction mixture was treated with 1NNa 2 CO 3 The crude product was obtained by washing once (4 mL) and once with saturated brine (4 mL), drying the organic phase and then spin-drying. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =100:1 to 25:1 to give 168-5 (95 mg, yield: 28.5%) as a brown solid, ESI-MS (m/z): 1006.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2- (benzofuran-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (168)
168-5 (95mg, 0.09mmol) was dissolved in ethyl acetate (5 mL), dioxane hydrochloride (4M, 2.5 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 1 hour. And (3) monitoring by TLC that most of raw materials are consumed, stopping the reaction, directly filtering the reaction solution, and drying the solid to obtain the product. Target compound 168 was a white solid (76 mg, yield: 87.9%), ESI-MS (m/z): 806.4[ 2 ] M + H] + 。
1 HNMR(400MHz,CD 3 OD,δ8.88-8.73(m,2H),8.07(m,1H),7.94(m,1H),7.57(m,1H),7.34-7.19(m,10H),4.70(m,1H),4.44-4.39(m,3H),4.14-3.98(m,3H),3.85-3.21(m,6H),3.03-2.93(m,4H),1.95-1.28(m,17H),0.99(d,3H),0.94(d,3H).
Example 4: synthesis of (R) -N- ((R) -6-amino-1- (1- (imidazo [1,2-a ] pyridine-2-carbonyl) -1, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trihydrochloride (198)
(1) Synthesis of 1- (imidazo [1,2-a ] pyridine-2-carbonyl) -1, 7-diazaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (198-2)
After dissolving 168-1 (0.19g, 1.2mmol), HATU (0.534g, 1.4mmol) and TEA (0.24g, 2.4mmol) in DMF (5 mL), the reaction mixture was stirred at room temperature for 30 minutes, 2 (0.23g, 1.0mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was used to monitor completion of the reaction, the reaction mixture was diluted with water (30 mL), extracted with DCM (10 mL. Times.3), and the organic phases were combined and then 1NNa was added 2 CO 3 Washed once (10 mL) and once with saturated brine (10 mL) and dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =100:1 to 30: 1A brown solid 198-2 (0.31 g, yield: 82.3%), ESI-MS (m/z): 371.1 2[ 2 ] M + H] + . (2) Imidazo [1,2-a ]]Pyridin-2-yl (1, 7-diazaspiro [3.5]]Synthesis of nonane-1-yl) methanone dihydrochloride (198-3)
198-2 (0.31g, 1.65mmol) was dissolved in THF (8 mL), dioxane hydrochloride solution (4M, 4 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 6 hours. And (4) monitoring by TLC that most of raw materials are completely consumed, stopping the reaction, and directly spin-drying the reaction solution to obtain the product. The objective compound 198-3 was a white solid (0.27 g, yield: 94.0%), ESI-MS (m/z): 271.0[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (1- (imidazo [1,2-a ] pyridine-2-carbonyl) -1, 7-diazaspiro [3.5] non-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (198-5)
After 1-1 (250mg, 0.33mmol), HATU (152mg, 0.40mmol) and DIEA (150mg, 1.16mmol) were dissolved in tetrahydrofuran (8 mL), the reaction mixture was stirred at room temperature for 30 minutes, and then added to the reaction mixture198-3 (123mg, 0.36mmol), the reaction was stirred at room temperature overnight, TLC monitored for completion, after which the reaction was run with 1NNa 2 CO 3 Washed once (3 mL) and once with saturated brine (3 mL) and dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =100:1 to 25:1 to obtain the objective compound 198-5 as a brown solid (75 mg, yield: 22.5%), ESI-MS (m/z): 1006.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (1- (imidazo [1,2-a ] pyridine-2-carbonyl) -1, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trihydrochloride (198)
198-5 (75mg, 0.07mmol) was dissolved in ethyl acetate (5 mL), and then dioxane hydrochloride (4M, 2.5 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 1 hour. And (3) monitoring by TLC that most of raw materials are consumed, stopping the reaction, directly filtering the reaction solution, and drying the solid to obtain the product. Target compound 198 as a white solid (52 mg, yield: 63.1%), ESI-MS (m/z): 806.3 2[ M ] +H] + 。
1 HNMR(400MHz,CD 3 OD),δ8.89-8.70(m,2H),8.04(m,1H),7.92(m,1H),7.53(m,1H),7.32-7.17(m,10H),4.70(m,1H),4.39(m,1H),4.15-4.73(m,5H),3.55(m,3H),3.36-3.13(m,2H),3.00-2.93(m,4H),2.37-2.19(m,4H),1.79-1.25(m,11H),0.96(d,3H),0.93(d,3H)。
Example 5: synthesis of (R) -N- ((R) -6-amino-1- (1- (2, 2-diphenylacetyl) -1, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (215)
(1) Synthesis of tert-butyl 1- (2, 2-diphenylacetyl) -1, 7-diazaspiro [3.5] nonane-7-carboxylate (215-3)
2 (50mg, 2.21X 10) -4 mol)，215-2(47mg，2.21×10 -4 mol)，HATU(126mg，3.31×10 - 4 mol) and triethylamine (86mg, 6.63X 10) -4 mol) was dissolved in tetrahydrofuran (10 mL), the reaction was stirred overnight at room temperature, TLC monitored for completion, after which the reaction was washed with 1NNa 2 CO 3 Washed once (5 mL) and once with 1N HCl (5 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V Ethyl acetate :V N-hexane =20:1 to 4:1 to give 215-3 as a pale yellow oil (60 mg, yield: 64.6%), ESI-MS (m/z): 421.2[ 2 ] M + H] + 。
(2) Synthesis of 2, 2-diphenyl-1- (1, 7-diazaspiro [3.5] nonan-1-yl) ethanone hydrochloride (215-4)
215-3 (60mg, 1.43X 10) -4 mol) was dissolved in DCM (6 mL), and dioxane hydrochloride (4M, 3 mL) was added to the solution, after which the reaction was stirred at room temperature for 2 hours. After the TLC monitoring reaction is completed, the reaction solution is directly dried by spinning to obtain the product. The aimed compound 215-4 was a pale yellow oil (50 mg, yield: 97.2%), ESI-MS (m/z): 321.2[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-Butoxycarbonylamino-1- (1- (2, 2-diphenylacetyl) -1, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (215-6)
1-1 (105mg, 1.39X 10) -4 mol)，HATU(79mg，2.09×10 -4 mol) and DIEA (72mg, 5.56X 10) -4 mol) was dissolved in tetrahydrofuran (10 mL), the reaction mixture was stirred at room temperature for 30 minutes, and 215-4 (50mg, 1.39X 10) -4 mol), the reaction was stirred at room temperature overnight, TLC monitored the completion of the reaction, after which the reaction was washed with 1NNa 2 CO 3 Washed once (5 mL) and once with 1N HCl (5 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =100:1 to 40:1 to give 215-6 (37 mg, yield: 26.7%) as a white solid, ESI-MS (m/z): 1056.6[ 2 ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (1- (2, 2-diphenylacetyl) -1, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (215)
215-6 (37mg, 3.70X 10) -5 mol) was dissolved in DCM (6 mL), and dioxane hydrochloride (4M, 3mL) was added to the solution, after which the reaction was stirred at room temperature for 2 hours. And stopping the reaction after the TLC monitoring reaction is completed, directly filtering the reaction solution, and drying the solid to obtain the product. The target compound 215 was a white solid (18 mg, yield: 52.4%), ESI-MS (m/z): 856.5[ M ] +H] + 。
Example 6: synthesis of (R) -N- ((R) -6-amino-1- (2- (isoindoline-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide bistrifluoroacetate (220)
(1) Synthesis of phenyl isoindoline-2-carboxylate (220-3)
After dissolving 220-1 (0.6 g, 3.86mmol) and triethylamine (0.97g, 9.65mmol) in tetrahydrofuran (20 mL), 220-2 (0.72g, 4.63mmol) was added to the solution in ice bath, the reaction was stirred overnight at room temperature, TLC monitored the reaction completion, the reaction was filtered, and the filtrate was directly spin-dried to give crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V N-hexane ：V Acetic acid ethyl ester =15:1 to 5:1 white solid 220-3 (0.55 g, yield: 59.6%), ESI-MS (m/z): 240.0 2[ M ] +H] + 。
(2) Synthesis of tert-butyl 2- (isoindoline-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (220-4)
After 220-3 (470mg, 1.97mmol) and 1 (490mg, 2.17mmol) were dissolved in DMSO (6 mL), K was added to the reaction mixture 2 CO 3 (816mg, 5.91mmol) and then the reaction mixture was stirred at 100 ℃ for 36 hours, and after completion of the reaction was monitored by TLC, the reaction mixture was poured into ice water (20 mL) and extracted with dichloromethane (10 mL. Times.3),the organic phases were combined and washed with saturated brine (10 mL. Times.2) and the organic phase was dried by spin-drying to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =80:1 to 20:1 to give 220-4 as a white solid (411 mg, yield: 56.3%), ESI-MS (m/z): 372.1[ 2 ] M + H] + 。
(3) Synthesis of Isofuran-2-yl (2, 7-diazepin [3,5] non-2-yl) methane trifluoroacetate (220-5)
After 220-4 (411mg, 1.11mmol) was dissolved in methylene chloride (6 mL), trifluoroacetic acid (2 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitoring most of the raw materials consumed, stop the reaction, directly spin-dry the reaction solution and slurry with ether, filter the solid and dry to obtain white solid compound 220-5 (415 mg, yield: 97.3%), ESI-MS (m/z): 272.1[ mu ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-tert-Butoxycarbonylamino-1- (2- (isoindoline-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-Butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (220-6)
After 1-1 (140mg, 0.19mmol), HATU (87mg, 0.23mmol) and DIEA (62mg, 0.48mmol) were dissolved in tetrahydrofuran (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 220-5 (77mg, 0.20mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was performed to monitor the completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed once (8 mL) and once with 1N HCl (8 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =80:1 to 25:1 to obtain the target compound 220-6 as a white solid (121 mg, yield: 64.6%), ESI-MS (m/z): 1007.5[ deg. ] M ] +H] + 。
(5) Synthesis of (R) -N- ((R) -6-amino-1- (2- (isoindoline-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide bistrifluoroacetate (220)
After dissolving 220-6 (121mg, 0.17mmol) in dichloromethane (4.5 mL), the solution was pouredAfter trifluoroacetic acid (1.5 mL) was added, the reaction was stirred at room temperature for 2 hours. TLC monitoring most of the raw materials are consumed, the reaction is stopped, the reaction solution is pulped by ether after being dried in a rotating way, and the solid is filtered and dried to obtain the product. Target compound 220 was a white solid (113 mg, yield: 90.9%), ESI-MS (m/z): 807.4[ M ] +H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.36-7.21(m,14H),4.73-4.70(m,6H),4.39(m,1H),4.09(m,1H),3.92-3.84(m,4H),3.72-3.63(m,2H),3.51-3.43(m,2H),3.27-3.19(m,2H),3.01-2.90(m,4H),1.86-1.29(m,13H),0.98(d,3H),0.94(d,3H).
Example 7: synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (E) -3- (thiophen-2-yl) acryloyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamido) -4-methylpentanamide dihydrochloride (229)
(1) Synthesis of (E) -2- (3- (thien-2-yl) acryloyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (229-3)
After dissolving 1 (150mg, 0.663mmol), 229-2 (102mg, 0.663mmol), HATU (378mg, 0.994mmol) and triethylamine (257mg, 1.99mmol) in tetrahydrofuran (10 mL), the reaction mixture was stirred at room temperature overnight, and after completion of the reaction was monitored by TLC, the reaction mixture was treated with 1NNa 2 CO 3 Washed once (3 mL) and once with 1N HCl (3 mL) respectively, dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V Ethyl acetate ：V N-hexane =20:1 to 4:1 to give 229-3 as a pale yellow oil (200 mg, yield: 83.2%), ESI-MS (m/z): 608.3[ 2 ] M + H] + 。
(2) Synthesis of (E) -1- (2, 7-diazaspiro [3.5] nonan-2-yl) -3- (thien-2-yl) prop-2-en-1-one hydrochloride (229-4)
229-3 (200mg, 0.552mmol) was dissolved in DCM (6 mL), and dioxane hydrochloride (4M, 3mL) was added to the solution, after which the reaction was stirred at room temperature for 2 hours. TLC monitoring of transAfter the reaction is completed, the reaction solution is directly spin-dried to obtain the product. Target compound 229-4 was a pale yellow oil (158 mg, yield: 96.0%), ESI-MS (m/z): 363.2[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1-oxo-1- (2- ((E) -3- (thien-2-yl) acryloyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (229-6)
After 1-1 (266mg, 0.353mmol), HATU (202mg, 0.530mmol) and DIEA (182mg, 1.41mmol) were dissolved in tetrahydrofuran (10 mL), the reaction mixture was stirred at room temperature for 30 minutes, 229-4 (158mg, 0.530mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC monitored the completion of the reaction, and then the reaction mixture was treated with 1NNa 2 CO 3 Washed once (3 mL) and once with 1N HCl (3 mL) respectively, dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =100:1 to 40:1 to obtain the aimed compounds 229-6 as white solids (195 mg, yield: 55.3%), ESI-MS (m/z): 998.5[ 2 ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (E) -3- (thiophen-2-yl) acryloyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamido) -4-methylpentanamide dihydrochloride (229)
229-6 (195mg, 1.95X 10) -5 mol) was dissolved in DCM (6 mL), and dioxane hydrochloride (4M, 3 mL) was added to the solution, after which the reaction was stirred at room temperature for 2 hours. And stopping the reaction after the TLC monitoring reaction is completed, directly filtering the reaction solution, and drying the solid to obtain the product. Target compound 229 was a white solid (120 mg, yield: 70.7%), ESI-MS (m/z): 798.4[ 2 ] M + H] + 。
1 H NMR(400MHz,CD 3 OD)δ8.00(s,1H),7.84–7.65(m,1H),7.55(t,J＝6.7Hz,1H),7.44–7.18(m,10H),7.16–7.00(m,1H),6.44(dd,J＝15.4,4.7Hz,1H),4.92–4.83(m,2H),4.74(dd,J＝9.8,4.8Hz,1H),4.42(d,J＝4.6Hz,1H),4.21–4.02(m,3H),3.94–3.59(m,7H),3.58–3.39(m,2H),3.25(dd,J＝13.9,5.3Hz,2H),3.08–2.79(m,8H),2.00–1.27(m,15H),0.99(dd,J＝15.2,6.2Hz,6H).
Example 8: synthesis of (R) -N- ((R) -6-amino-1- (2-cinnamoyl-2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide dihydrochloride (230)
(1) Synthesis of 2-cinnamoyl-2, 7-diazaspiro [3.5] nonane-7-carboxylic acid tert-butyl ester (230-2)
1 (250mg, 1.105mmol), 230-1 (136.3mg, 0.921mmol), HOBT (149mg, 1.105mmol) and EDC. HCl (264.7mg, 1.38mmol) were dissolved in methylene chloride (5 mL), stirred at room temperature for 5 minutes, triethylamine (93mg, 0.92mmol) was added, and the reaction was stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate (5 mL), the organic phase was separated, the aqueous phase was extracted with dichloromethane (5 mL. Times.3), the organic phases were combined, washed with 1N dilute hydrochloric acid (5 mL), and concentrated to give crude 230-2 (251 mg, yield: 76.7%), ESI-MS (m/z): 357.2[ M ] +H] + 。
(2) Synthesis of (E) -3-phenyl-1- (2, 7-diazaspiro [3.5] nonan-2-yl) prop-2-en-1-one hydrochloride (230-3)
After the crude 230-2 (251mg, 0.71mmol) was dissolved in 5mL of ethyl acetate, 2.5mL of dioxane hydrochloride solution was added, and the reaction was stirred at room temperature for 2 hours and filtered to obtain 230-3 (184 mg, yield: 89.2%) as a white solid, ESI-MS (m/z): 257.2[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (2-cinnamyl-2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (230-4)
After 1-1 (300mg, 0.398mmol) and HATU (90.8mg, 0.239mmol) were dissolved in THF (5 mL), DIEA (102.8mg, 0.797mmol) was added thereto, and the mixture was stirred at room temperature for 0.5 hour, then 230-3 (70mg, 0.239mmol) was added thereto, and the mixture was stirred at room temperature overnight. Adding saturated aqueous sodium bicarbonate solution (5 mL) of the reaction mixture, the organic phase was separated and the aqueous phase was quenched with V DCM ：V MeOH Extraction of =10 (5 mL × 3), merging of organic phases, washing with 1N diluted hydrochloric acid (5 mL), concentration, and column chromatography (V) DCM :V MeOH = 20): 48.6%), ESI-MS (m/z): 992.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2-cinnamoyl-2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamido) -4-methylpentanamide dihydrochloride (230)
After 230-4 (192mg, 0.19mmol) was dissolved in 5mL of ethyl acetate, 2.5mL of dioxane hydrochloride solution was added, and the mixture was stirred at room temperature for 2 hours and then vacuum-filtered to obtain a brown solid compound 230 (128 mg, yield: 76.5%), ESI-MS (m/z): 792.4[ M ] +H] + 。
1 HNMR(400MHz,CD 3 OD)δ7.65-7.52(m,3H),7.44-7.24(m,13H),6.77-6.67(m,1H),4.75-4.72(m,1H),4.46-4.41(m,1H),4.20-4.11(m,2H),3.90-3.80(m,2H),3.26-3.23(m,4H),3.04-2.89(m,8H),1.89-1.47(m,13H),0.99(dd,6H).
Example 9: synthesis of (R) -N- ((R) -6-amino-1- (2- ((E) -3- (furan-3-yl) acryloyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionylamino) -4-methylpentanamide dihydrochloride (239)
(1) Synthesis of (E) -tert-butyl 2- (3- (furan-3-yl) acrylate) -2, 7-diazepine [3.5] non-7-carboxylate (239-2)
1 (250mg, 1.105mmol), 239-1 (127mg, 0.921mmol), HOBT (149mg, 1.105mmol) and EDC. HCl (264.7 mg, 1.38mmol) were dissolved in 5mL of dichloromethane, stirred at room temperature for 5 minutes, added with triethylamine (93mg, 0.92mmol), and stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate (5 mL), the organic phase was separated, the aqueous phase was extracted with dichloromethane (5 mL. Times.3), the organic phases were combined, washed with 1N dilute hydrochloric acid (5 mL)And concentrated to give crude 239-2 (274 mg, yield: 43.7%), ESI-MS (m/z): 347.2[ 2 ] M + H] + 。
(2) Synthesis of (E) -3- (furan-3-yl) -1- (2, 7-diazepin [3.5] non-2-yl) prop-2-en-1-one hydrochloride (239-3)
Dissolving the crude product 239-2 (274mg, 0.79mmol) with 5mL ethyl acetate, adding 2.5mL dioxane hydrochloride solution, stirring at room temperature for reaction for 2 hours, filtering to obtain white solid 239-3 (186 mg, yield: 83.1%), ESI-MS (m/z): 247.2M + H +] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (2- ((E) -3- (furan-3-yl) acryloyl) -2, 7-diazaspiro [3.5] non-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (239-4)
After 1-1 (300mg, 0.398mmol) and HATU (90.8mg, 0.239mmol) were dissolved in THF (5 mL), DIEA (102.8mg, 0.797mmol) was added thereto, and the mixture was stirred at room temperature for 0.5 hour, then 239-3 (67mg.0.239mmol) was added thereto, and the mixture was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (5 mL), the organic phase separated and the aqueous phase diluted with V DCM ：V MeOH Extraction of =10 (5 mL × 3), merging of organic phases, washing with 1N diluted hydrochloric acid (5 mL), concentration, and column chromatography (V) DCM :V MeOH = 20): 53.0%), ESI-MS (m/z): 982.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2- ((E) -3- (furan-3-yl) acryloyl) -2, 7-diazaspiro [3.5] non-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionylamino) -4-methylpentanamide dihydrochloride (239)
After 239-4 (207mg, 0.21mmol) was dissolved in 5mL of ethyl acetate, 2.5mL of dioxane hydrochloride was added, the reaction was stirred at room temperature for 2 hours, and suction filtration was carried out to obtain a brown solid compound 239 (121 mg, yield: 67.1%), ESI-MS (m/z): 782.4[ M ] +H] + 。
1 H NMR(400MHz,CD 3 OD)δ7.81(s,1H),7.58(s,1H),7.55-7.52(m,1H),7.39-7.22(m,10H),6.83(s,1H),6.47-6.43(m,1H),4.75-4.72(m,1H),4.46-4.41(m,1H),4.20-4.11(m,2H),3.90-3.80(m,1H),3.26-3.23(m,4H),3.04-2.89(m,9H),1.89-1.47(m,13H),0.99(dd,6H).
Example 10: synthesis of (R) -N- ((R) -6-amino-1- (2- (imidazo [1,2-a ] pyridine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -4-methyl-2- ((R) -3-phenyl-2- (2- ((R) -2-phenylpropyl) amino) acetamido) pentanamide trihydrochloride (240)
(1) Synthesis of tert-butyl ((10R, 13R, 16R) -16-benzyl-10- (2- (imidazo [1,2-a ] pyridine-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carbonyl) -13-isobutyl-2, 2-dimethyl-4, 12, 15, 18-tetraoxo-3-oxa-5, 11, 14, 17-tetradecazanonan-19-yl) ((R) -2-phenylpropyl) carbamate (240-1)
After 1-2 (60mg, 0.077mmol), HATU (35mg, 0.092mmol) and DIEA (30mg, 0.231mmol) were dissolved in DMF (3 mL), the reaction mixture was stirred at room temperature for 30 minutes, 168-3 (22mg, 0.081mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, after completion of the reaction was monitored by TLC, the reaction mixture was directly purified by a reverse phase column to give 240-5 (42 mg, yield: 52.9%) as a brown solid, ESI-MS (m/z): 1034.4[ M ] of calcium chloride] + 。
(2) Synthesis of (R) -N- ((R) -6-amino-1- (2- (imidazo [1,2-a ] pyridine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -4-methyl-2- ((R) -3-phenyl-2- (2- ((R) -2-phenylpropyl) amino) acetamido) pentanamide trihydrochloride (240)
After 240-1 (42mg, 0.041mmol) was dissolved in ethyl acetate (3 mL), dioxane hydrochloride (4M, 1.5mL) was added to the solution, and then the reaction mixture was stirred at room temperature for 1 hour. And (3) monitoring by TLC that most of raw materials are consumed, stopping the reaction, directly filtering the reaction solution, and drying the solid to obtain the product. Target compound 240 was a white solid (29 mg, yield: 76.4%), ESI-MS (m/z): 834.3[ M ] +H] + 。
1 HNMR(400MHz,CD 3 OD),δ8.87-8.73(m,2H),8.07(m,1H),7.93(m,1H),7.57(m,1H),7.37-7.14(m,10H),4.70(m,1H),4.55(m,1H),4.47-4.39(m,3H),4.12-4.00(m,4H),3.88-3.72(m,2H),3.59-3.46(m,2H),3.23-2.83(m,7H),2.01-1.31(m,16H),0.97(d,3H),0.91(d,3H).
Example 11: synthesis of (R) -N- ((R) -6-amino-1- (2- (3, 3-difluoroazetidine-1-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide bistrifluoroacetate (201)
(1) Synthesis of tert-butyl 2- (3, 3-difluoroazetidine-1-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (201-2)
After 1 (100mg, 0.442mmol) and triphosgene (52.5mg, 0.177mmol) were dissolved in 3mL of dichloromethane, triethylamine (89mg, 0.884mmol) was added thereto, and the reaction was stirred at room temperature for 3 hours. The reaction was quenched with water (20 mL), the organic phase separated, the aqueous phase extracted with dichloromethane (5 mL. Times.3), the organic phases combined and concentrated.
After 201-1 (57.4 mg, 0.442mmol) was dissolved in 3mL of dichloromethane, triethylamine (89mg, 0.884mmol) and the DCM solution obtained in the previous step were added and the reaction was stirred at room temperature overnight. Quenching the reaction with water, separating the organic phase, extracting the aqueous phase with dichloromethane (10 mL. Times.3), combining the organic phases, concentrating, and performing column chromatography (V) DCM :V MeOH =20, 1) to give 201-2 as a white solid (130 mg, yield: 85.2%), ESI-MS (m/z): 346.2[ M ] +H] + 。
(2) Synthesis of (3, 3-difluoroazetidin-1-yl) (2, 7-diazaspiro [3.5] nonan-2-yl) methanone trifluoroacetate (201-3)
After dissolving 201-2 (130mg, 0.38mmol) in 5mL of dichloromethane, 2.5mL of trifluoroacetic acid was added, and the reaction was stirred at room temperature for 2 hours and concentrated to obtain 201-3 (115 mg, yield: 85.0%), ESI-MS (m/z): 246.2[ 2 ] M + H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1- (2- (3, 3-difluoroazetidine-1-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (201-4)
After 1-1 (150mg, 0.199mmol) and HATU (90.8mg, 0.239mmol) were dissolved in THF (5 mL), DIEA (102.8mg, 0.797mmol) was added, and the reaction mixture was stirred at room temperature for 0.5 hour, then 201-3 (85.8mg.0.239mmol) was added, and the reaction mixture was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (5 mL), the organic phase separated and the aqueous phase diluted with V DCM ：V MeOH Extraction of =10 (5 mL × 3), organic phases combined, washed with 1N dilute hydrochloric acid (5 mL), concentrated, and column chromatographed (V) DCM :V MeOH = 20): 54.4%), ESI-MS (m/z): 981.5[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1- (2- (3, 3-difluoroazetidine-1-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropanamido) -4-methylpentanamid bistrifluoroacetate (201)
After 201-4 (106mg, 0.11mmol) was dissolved in 5mL of dichloromethane, 2.5mL of trifluoroacetic acid was added, the reaction was stirred at room temperature for 2 hours, concentrated, and a solid was precipitated by addition of ether and suction-filtered to obtain a white solid 201 (91 mg, yield: 83.5%), ESI-MS (m/z): 781.4[ M ] +H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.40-7.24(m,10H),4.78-4.72(m,1H),4.47-4.27(m,5H),4.13-4.07(m,1H),3.81-3.63(m,8H),3.04-2.89(m,7H),1.89-1.34(m,13H),0.99(dd,6H).
Example 12: synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -5-oxopyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide bistrifluoroacetate (337)
(1) Synthesis of tert-butyl (R) -2- (5-oxopyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (337-2)
1 (210mg, 0.9 mmol), 337-1 (100mg, 0.775mmol), HOBT (125mg, 0.93mmol) and EDC. HCl (223mg, 1.163mmol) were dissolved in 5mL of dichloromethane, stirred at room temperature for 5 minutes, added triethylamine (78mg, 0.775mmol) and stirred at room temperature overnight. The reaction was quenched with saturated aqueous sodium bicarbonate (5 mL), the organic phase was separated, the aqueous phase was extracted 3 times with dichloromethane, the organic phases were combined, washed with 1N diluted hydrochloric acid, and concentrated to give crude 337-2 (214 mg, yield: 81.9%), ESI-MS (m/z): 338.1[ M ] +H] + 。
(2) Synthesis of (R) -5- (2, 7-diazaspiro [3.5] nonane-2-carbonyl) pyrrolidin-2-one (337-3)
The crude 337-2 (214mg, 0.63mmol) was dissolved in 5mL of dichloromethane, and then 2.5mL of trifluoroacetic acid was added thereto, followed by stirring at room temperature for 2 hours, followed by filtration to obtain a white solid 337-3 (205 mg, yield: 92.0%), ESI-MS (m/z): 238.1[ 2 ] M + H] + 。
(3) Synthesis of (R) -N- ((R) -6-tert-Butoxycarbonylamino-1-oxo-1- (2- ((R) -5-oxopyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (337-4)
After 1-1 (200mg, 0.265mmol) and HATU (121mg, 0.319mmol) were dissolved in THF (5 mL), DIEA (137mg, 1.062mmol) was added, and after stirring at room temperature for 0.5 hour, 337-3 (112mg, 0.319mmol) was added, and the reaction was stirred at room temperature overnight. The reaction was quenched by addition of saturated aqueous sodium bicarbonate (5 mL), the organic phase separated and the aqueous phase diluted with V DCM ：V MeOH Extraction of =10 (5 mL × 3), merging of organic phases, washing with 1N diluted hydrochloric acid (5 mL), concentration, and column chromatography (V) DCM :V MeOH = 20): 51.9%), ESI-MS (m/z): 973.5[ deg. ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -5-oxopyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide bistrifluoroacetate (337)
337-4 (134mg, 0.14mmol) was incorporated into 5mL of methylene chlorideAfter the alkane was dissolved, 1mL of trifluoroacetic acid was added, and the reaction was stirred at room temperature for 2 hours and suction-filtered to obtain a white solid 337 (90 mg, yield: 65.3%), ESI-MS (m/z): 773.5[ 2 ] M + H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.36-7.22(m,10H),4.76-4.67(m,1H),4.44-4.29(m,2H),4.15-4.05(m,3H),3.80-3.64(m,4H),3.49-3.42(m,2H),3.19-2.84(m,5H),2.47-2.29(m,3H),2.13-2.04(m,2H),1.88-1.40(m,14H),0.99(dd,6H).
Example 13: synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -1,2,3, 4-tetrahydroisoquinoline-3-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylalanyl) -4-methylpentanamide trihydrochloride (264)
(1) Synthesis of (R) - (9H-fluoren-9-yl) methyl 3- (7- (tert-butoxycarbonyl) -2, 7-diazaspiro [3.5] nonane-2-carbonyl) -3, 4-dihydroisoquinoline-2 (1H) -carboxylate (264-3)
After dissolving 1 (200mg, 0.884mmol), 264-2 (353mg, 0.884mmol), HATU (403mg, 1.06mmol) and DIEA (228mg, 1.77mmol) in tetrahydrofuran (10 mL), the reaction mixture was stirred overnight at room temperature, and after completion of the reaction was monitored by TLC, the reaction mixture was treated with 1NNa 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V Ethyl acetate ：V N-hexane =20:1 to 4:1 to give 264-3 (400 mg, yield: 74.5%) as a pale yellow oil, ESI-MS (m/z): 608.3[ 2 ] M + H] + 。
(2) Synthesis of (R) - (9H-fluoren-9-yl) methyl 3- (2, 7-diazaspiro [3.5] nonane-2-carbonyl) -3, 4-dihydroisoquinoline-2 (1H) -carboxylic acid ester hydrochloride (264-4)
264-3 (400mg, 0.658mmol) was dissolved in DCM (6 mL), and then dioxane hydrochloride (4M, 3mL) was added to the solution, followed by stirring at room temperature for 2 hours. After the TLC monitoring reaction is completed, the reaction solution is directly dried by spinning to obtain the product. Target compound264-4 as a pale yellow oil (356 mg, yield: 99.5%), ESI-MS (m/z): 508.3[ 2 ] M + H] + 。
(3) Synthesis of (R) - (9H-fluoren-9-yl) methyl 3- (7- ((6R, 9R,12R, 15R) -6, 9-dibenzyl-15- (4- (tert-butoxycarbonyl) amino) butyl) -12-isobutyl-2, 2-dimethyl-4, 7, 10, 13-tetraoxo-3-oxa-5, 8, 11, 14-tetraazahexadeca-16-acyl) -2, 7-diazaspiro [3.5] non-2-carbonyl) -3, 4-dihydroisoquinoline-2 (1H) -carboxylate (264-6)
After 1-1 (150mg, 0.199mmol), HATU (113mg, 0.298m mol) and DIEA (154mg, 0.796 mmol) were dissolved in tetrahydrofuran (10 mL), the reaction mixture was stirred at room temperature for 30 minutes, 264-4 (151mg, 0.298mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, and after completion of the reaction was monitored by TLC, the reaction mixture was treated with 1N Na 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =100:1 to 40:1 to obtain the target compound 264-6 as a white solid (135 mg, yield: 54.5%), ESI-MS (m/z): 1243.7[ 2 ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-tert-butoxycarbonylamino-1-oxo-1- (2- ((R) -1,2,3, 4-tetrahydroisoquinoline-3-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (264-7)
264-6 (135mg, 0.109mmol) was dissolved in dichloromethane (6 mL), piperidine (3 mL) was added, the mixture was stirred overnight at room temperature, and the reaction was monitored by TLC and then spun dry. Purifying by column chromatography, eluting with polar solvent V DCM ：V MeOH =100:1 to 20:1 to obtain the target compound 264-7 as a white solid (100 mg, yield: 89.8%), ESI-MS (m/z): 1021.6[ 2 ] M + H] + 。
(5) Synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -1,2,3, 4-tetrahydroisoquinoline-3-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylamido) -4-methylpentanamide trihydrochloride (264)
Mixing 264-7 (100mg, 9.79X 10) -5 mol) was dissolved in DCM (6 mL), and dioxane hydrochloride (4M, 3mL) was added to the solution, after which the reaction was stirred at room temperature for 2 hours. And stopping the reaction after the TLC monitoring reaction is completed, directly filtering the reaction solution, and drying the solid to obtain the product. The target compound 264 was a white solid (88 mg, yield: 96.6%), ESI-MS (m/z): 821.5[ M ] +H] + 。
1 H NMR(400MHz,CD 3 OD)δ7.55–7.14(m,14H),4.73(s,1H),4.61–4.02(m,6H),4.00–3.62(m,4H),3.61–3.35(m,4H),3.29–2.83(m,9H),2.10–1.28(m,17H),1.11–0.82(m,6H).
Example 14: synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -piperidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trihydrochloride (265)
(1) Synthesis of (R) -tert-butyl 2- (1- (((9H-fluoren-9-yl) methoxy) carbonyl) piperidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylic acid ester (265-3)
After dissolving 1 (200mg, 0.884mmol), 265-2 (311mg, 0.884mmol), HATU (403mg, 1.06mmol) and DIEA (228mg, 1.77mmol) in tetrahydrofuran (10 mL), the reaction was stirred overnight at room temperature, and after completion of the reaction was monitored by TLC, the reaction was monitored by 1NNa 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V Acetic acid ethyl ester ：V N-hexane =20:1 to 4:1 to give 265-3 as a pale yellow oil (460 mg, yield: 93.0%), ESI-MS (m/z): 560.3[ 2 ] M + H] + 。
(2) Synthesis of (R) - (9H-fluoren-9-yl) methyl 2- (2, 7-diazaspiro [3.5] nonane-2-carbonyl) piperidine-1-carboxylate (265-4)
265-3 (460mg g, 0.822mmol) was dissolved in DCM (6 mL), dioxane hydrochloride solution (4M, 3mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLCAnd (5) monitoring the reaction, and directly spin-drying the reaction solution to obtain the product. Target compound 265-4 was a pale yellow oil (406 mg, yield: 99.5%), ESI-MS (m/z): 460.3 2[ M ] +H] + 。
(3) Synthesis of (R) - (9H-fluoren-9-yl) methyl 2- (7- ((6R, 9R,12R, 15R) -6, 9-dibenzyl-15- (4- (tert-butoxycarbonyl) amino) butyl) -12-isobutyl-2, 2-dimethyl-4, 7, 10, 13-tetraoxo-3-oxa-5, 8, 11, 14-tetraazahexadec-16-acyl) -2, 7-diazaspiro [3.5] nonane (265-6)
After dissolving 1-1 (100mg, 0.133mmol), HATU (76mg, 0.199mmol) and DIEA (69mg, 0.532mmol) in tetrahydrofuran (10 mL), the reaction solution was stirred at room temperature for 30 minutes, 265-4 (99mg, 0.199mmol) was added to the reaction solution, the reaction solution was stirred at room temperature overnight, after monitoring the reaction by TLC, the reaction solution was then treated with 1NNa 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =100:1 to 40:1 to give 265-6 (50 mg, yield: 31.4%) as a white solid, ESI-MS (m/z): 1195.7[ mu ] M + H] + 。
(4) Synthesis of (R) -N- ((R) -6-tert-Butoxycarbonylamino-1-oxo-1- (2- ((R) -piperidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylamido) -4-methylpentanamide (265-7)
Mixing 265-6 (50mg, 4.18X 10) -5 mol), dissolved in dichloromethane (6 mL), piperidine (3 mL) was added, stirred at room temperature overnight, the reaction was monitored by TLC and, after completion of the reaction, was spun dry. Purifying by column chromatography, eluting with polar solvent V DCM ：V MeOH =100:1 to 20:1 to give 265-7 (30 mg, yield: 73.7%) as a white solid, ESI-MS (m/z): 973.6[ M ] +1] + 。
(5) Synthesis of (R) -N- ((R) -6-amino-1-oxo-1- (2- (R) -piperidine-2-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) hex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trihydrochloride (265)
265-7 (30mg, 3.08X 10) -5 mol) dissolved in DCM (6 mL) beforeDioxane hydrochloride solution (4M, 3mL) was added to the solution, and then the reaction solution was stirred at room temperature for 2 hours. And stopping the reaction after the TLC monitoring reaction is completed, directly filtering the reaction solution, and drying the solid to obtain the product. Target compound 265 as a white solid (25 mg, yield: 92.0%), ESI-MS (m/z): 773.5[ 2 ] M + H] + 。
1 H NMR(400MHz,CD 3 OD)δ7.53–7.16(m,10H),4.71(dt,J＝18.4,9.1Hz,1H),4.41(dt,J＝9.8,5.0Hz,1H),4.26–3.96(m,4H),3.94–3.59(m,4H),3.58–3.34(m,4H),3.29–2.76(m,8H),2.27–1.35(m,23H),0.98(dd,J＝16.0,6.2Hz,6H).
Example 15: synthesis of 7- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamido) -4-methylpentanoylamino) hexanoyl) -N- (3-fluoro-4- (trifluoromethoxy) phenyl) -2, 7-diazaspiro [3.5] nonane-2-carboxamide ditrifluoroacetate (253)
(1) Synthesis of phenyl (3-fluoro-4- (trifluoromethoxy) phenyl) carbamate (253-2)
253-1 (0.45g, 2.31mmol) and pyridine (0.36g, 4.62mmol) were dissolved in tetrahydrofuran (16 mL), and 220-2 (0.43g, 2.77mmol) was added to the solution under ice bath, after which the reaction was stirred at room temperature overnight, TLC monitored for completion of the reaction, filtered, and the filtrate was directly spin-dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V N-hexane ：V Ethyl acetate =15:1 to 5:1 to give 253-2 as a white solid (0.70 g, yield: 96.3%), ESI-MS (m/z): 316.0[ M ] +H] + 。
(2) Synthesis of tert-butyl 2- (3-fluoro-4- (trifluoromethoxy) phenyl) carbamoyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (253-3)
253-2 (700mg, 2.22mmol) and 1 (551mg, 2.44mmol) were dissolved in DMSO (6 mL), and K was added to the reaction mixture 2 CO 3 (1.22g, 8.88mmol) after which the reaction was stirred at room temperature overnight, TLC monitored the reaction was complete and poured into ice water (30 mL)After extraction with dichloromethane (10 mL. Times.3), the organic phases were combined and washed twice with saturated brine (10 mL), dried and spin-dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =80:1 to 20:1 to give 253-3 as a white solid (779 mg, yield: 78.4%), ESI-MS (m/z): 448.1[ 2 ] M + H] + 。
(3) Synthesis of N- (3-fluoro-4- (trifluoromethoxy) phenyl) -2, 7-diazaspiro [3.5] nonane-2-carboxamide trifluoroacetate (253-4)
After 253-3 (250mg, 0.56mmol) was dissolved in methylene chloride (6 mL), trifluoroacetic acid (2 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitored most of the starting material was consumed, the reaction was stopped, the reaction solution was spun dry and slurried with diethyl ether (10 mL), filtered, and the solid was dried to give 253-4 as a white compound (248 mg, yield: 96.2%), ESI-MS (m/z): 348.1 2[ M ] +H] + 。
(4) Synthesis of 7- ((R) -6-tert-butoxycarbonylamino-2- ((R) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionamino) -4-methylpentanoylamino) hexanoyl) -N- (3-fluoro-4- (trifluoromethoxy) phenyl) -2, 7-diazaspiro [3.5] nonane-2-carboxamide (253-6)
After 1-1 (140mg, 0.19mmol), HATU (87mg, 0.23mmol) and DIEA (62mg, 0.48mmol) were dissolved in tetrahydrofuran (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 253-4 (92mg, 0.20mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was performed to monitor the completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), and the organic phase dried and spun to the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =80:1 to 30:1 to give 253-6 as a white solid (121 mg, yield: 60.1%), ESI-MS (m/z): 1083.3[ 2 ] M + H] + 。
(5) Synthesis of 7- ((R) -6-amino-2- ((R) -2- ((R) -2- ((R) -2-amino-3-phenylpropionamino) -4-methylpentanoylamino) hexanoyl) -N- (3-fluoro-4- (trifluoromethoxy) phenyl) -2, 7-diazaspiro [3.5] nonane-2-carboxamide ditrifluoroacetate (253)
After 253-6 (121mg, 0.17mmol) was dissolved in methylene chloride (4.5 mL), trifluoroacetic acid (1.5 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitoring most raw materials are completely consumed, the reaction is stopped, the reaction liquid is dehydrated and then pulped by ether (20 mL), the solid is filtered and dried to obtain the product. Target compound 253 as a white solid (95 mg, yield: 76.5%), ESI-MS (m/z): 883.2[ M ] C + H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.62(m,2H),7.37-7.21(m,11H),4.86(m,1H),4.71(m,1H),4.38(m,1H),4.08(m,1H),3.85-3.63(m,6H),3.51-3.38(m,2H),3.26-3.19(m,2H),3.00-2.90(m,4H),1.88-1.42(m,13H)，0.98(d,3H),0.94(d,3H).
Example 16: synthesis of (2R) -N- ((2R) -6-amino-1- (2- (hexahydro-1H-furo [3,4-c ] pyrrole-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylamido) -4-methylpentanamide ditrifluoroacetate (256)
(1) Synthesis of 7-tert-butyl 2-phenyl 2, 7-diazaspiro [3.5] nonane-2, 7-dicarboxylate (256-2)
After 1 (0.5g, 2.21mmol) and triethylamine (0.45g, 4.42mmol) were dissolved in tetrahydrofuran (20 mL), 220-2 (0.41g, 2.65mmol) was added to the above solution under ice bath, the reaction solution was stirred at room temperature overnight, TLC was used to monitor completion of the reaction, and the filtrate was directly spin-dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V N-hexane ：V Acetic acid ethyl ester =15:1 to 5:1 to give 256-2 as a white solid (0.72 g, yield: 94.1%), ESI-MS (m/z): 347.1[ 2 ] M + H] + 。
(2) Synthesis of tert-butyl 2- (hexahydro-1H-furo [3,4-c ] pyrrole-5-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (256-3)
Mixing 256-2 (660mg, 1.91mmol) and 6H-1H-furan [3,4-C]After pyrrole hydrochloride (314mg, 2.10mmol) was dissolved in DMSO (6 mL), K was added to the reaction mixture 2 CO 3 (791mg, 5.73mmol), the reaction was stirred at 100 ℃ for 16 hours, TLC monitored completion, poured into ice water (30 mL), extracted with dichloromethane (10 mL. Times.3), and the organic phases were combined and washed twice with saturated brine (10 mL) and dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =80:1 to 25:1 to give 256-3 (405 mg, yield: 58.2%) as a white solid, ESI-MS (m/z): 366.1[ 2 ] M + H] + 。
(3) Synthesis of 2, 7-diazaspiro [3.5] nonan-2-yl (tetrahydro-1H-furo [3,4-c ] pyrrol-5 (3H) -yl) methanone (256-4)
After 256-3 (405mg, 1.11mmol) was dissolved in dichloromethane (6 mL), trifluoroacetic acid (2 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitored most of the starting material was consumed, the reaction was stopped, the reaction solution was directly spun dry and slurried with diethyl ether, filtered, and the solid was dried to give 256-4 (401 mg, yield: 95.3%) as a white solid, ESI-MS (m/z): 266.1[ M ] +H] + 。
(4) Synthesis of (2R) -N- ((2R) -6-tert-Butoxycarbonylamino-1- (2- (hexahydro-1H-furo [3,4-c ] pyrrole-5-carbonyl) -2, 7-diazaspiro [3.5] non-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (256-6)
After 1-1 (140mg, 0.19mmol), HATU (87mg, 0.23mmol) and DIEA (62mg, 0.48mmol) were dissolved in tetrahydrofuran (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 256-4 (76mg, 0.20mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was performed to monitor completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =80:1 to 25:1 to give 256-6 (127 mg, yield: 68.2%) as a white solid, ESI-MS (m/z): 1001.4[ 2 ] M + H] + 。
(5) Synthesis of (2R) -N- ((2R) -6-amino-1- (2- (hexahydro-1H-furo [3,4-c ] pyrrole-5-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylalanyl) -4-methylpentanamide ditrifluoroacetate (256)
After 256-6 (127mg, 0.13mmol) was dissolved in methylene chloride (4.5 mL), trifluoroacetic acid (1.5 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitoring most of raw materials are consumed, the reaction is stopped, the reaction solution is dehydrated, then is pulped by ether (10 mL), filtered and dried to obtain the product. The target compound 256 was a white solid (122 mg, yield: 93.4%), ESI-MS (m/z): 801.3[ 2 ] M + H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.39-7.21(m,10H),4.88(m,1H),4.73(m,1H),4.41(m,1H),4.11(m,1H),3.90(m,1H),3.83-3.58(m,10H),3.52-3.43(m,3H),3.29-3.21(m,2H),3.03-2.92(m,6H),1.43-1.45(m,13H)，0.99(d,3H),0.96(d,3H).
Example 17: synthesis of (2R) -N- ((2R) -6-amino-1- (2- (octahydrocyclopenta [ c ] pyrrole-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- (R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trifluoroacetate (343)
(1) Synthesis of tert-butyl 2- (octahydrocyclopenta [ c ] pyrrole-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (343-3)
After 256-2 (400mg, 1.116mmol) and 343-2 (154mg, 1.39mmol) were dissolved in DMSO (6 mL), K was added to the reaction mixture 2 CO 3 (800mg, 5.8 mmol) the reaction was stirred at 105 ℃ for 16 h, TLC monitored for completion, poured onto ice water (30 mL), extracted with dichloromethane (10 mL. Times.3), and the organic phases were combined and washed twice with saturated brine (10 mL), dried and spin-dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =60:1 to 20:1 to give 343-3 (305 mg, yield: 71.7%) as a pale brown solid, ESI-MS (m/z): 364.1 2[ 2 ] M + H] + 。
(2) Synthesis of (hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl) (2, 7-diazaspiro [3.5] nonan-2-yl) methanone (343-4)
After 343-3 (305mg, 0.85mmol) was dissolved in methylene chloride (12 mL), trifluoroacetic acid (4 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. TLC monitored most of the starting material consumption, stopped the reaction, spun the reaction solution directly and slurried with ether (10 mL), filtered, and dried to give target compound 343-4 as a white solid (270 mg, yield: 85.2%), ESI-MS (m/z): 264.1[ M ] +H] + 。
(3) Synthesis of (2R) -N- ((2R) -6-tert-butoxycarbonylamino-1- (2- (octahydrocyclopenta [ c ] pyrrole-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (343-6)
After 1-1 (150mg, 0.20mmol), HATU (91mg, 0.24mmol) and DIEA (77mg, 0.60mmol) were dissolved in tetrahydrofuran (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 343-4 (83mg, 0.22mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was performed to monitor completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed once (10 mL) and once with 1N HCl (10 mL), dried the organic phase and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM :V MeOH =60:1 to 20:1 to give 343-6 (117 mg, yield: 58.9%) as a white solid, ESI-MS (m/z): 999.4[ M ] +H] + 。
(4) Synthesis of (2R) -N- ((2R) -6-amino-1- (2- (octahydrocyclopenta [ c ] pyrrole-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -1-oxohex-2-yl) -2- ((R) -2- (R) -2-amino-3-phenylpropionamino) -4-methylpentanamide trifluoroacetate (343)
After 343-6 (117mg, 0.12mmol) was dissolved in methylene chloride (4.5 mL), trifluoroacetic acid (1.5 mL) was added to the solution, and the reaction mixture was stirred at room temperature for 2 hours. And (3) monitoring by TLC that most of raw materials are completely consumed, stopping the reaction, pulping the reaction solution by using diethyl ether after the reaction solution is dried in a rotating way, filtering and drying to obtain the product. Target compound 343 was a white solid (107 mg, yield: 89.0%), ESI-MS (m/z): 801.2[M+H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.37-7.23(m,10H),4.72(m,1H),4.39(m,1H),4.10(m,1H),3.81-3.41(m,9H),3.31-3.17(m,4H),3.09-2.92(m,4H),2.67(m,2H),1.85-1.45(m,21H)，0.99(d,3H),0.96(d,3H).
Example 18: synthesis of (R) -N- ((R) -1- (2- ((R) -1-acetylpyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -6-amino-1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropanamido) -4-methylpentanamid ditrifluoroacetate (354)
(1) Synthesis of tert-butyl (R) -2- (1-acetylpyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonane-7-carboxylate (354-2)
After 1 (400mg, 1.77mmol), HATU (806mg, 2.12mmol) and DIEA (457mg, 3.54mmol) were dissolved in tetrahydrofuran (12 mL), the reaction mixture was stirred at room temperature for 30 minutes, 354-1 (306mg, 1.95mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was used to monitor completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed separately (10 mL) and 1N HCl (10 mL), and the organic phase was dried and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =80:1 to 30:1 yellow oil 354-2 (611 mg, yield: 94.6%), ESI-MS (m/z): 366.1[ M ] +H] + 。
(2) Synthesis of (R) -1- (2, 7-diazaspiro [3.5] nonane-2-carbonyl) pyrrolidin-1-yl) ethanone trifluoroacetate (354-3)
354-2 (611mg, 1.54mmol) was dissolved in DCM (12 mL), and trifluoroacetic acid (4 mL) was added to the solution, after which the reaction was stirred at room temperature for 3 hours. TLC monitored most of the starting material consumption, stopped the reaction, dried the reaction solution by spinning and slurrying with diethyl ether (10 mL), filtered, and the solid dried to give 354-3 (594 mg, yield: 93.6%) as a white solid, ESI-MS (m/z): 266.1[ M ] +H] + 。
(3) Synthesis of (R) -N- ((R) -1- (2- ((R) -1-acetylpyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -6-tert-butoxycarbonylamino-1-oxohex-2-yl) -2- ((R) -2- ((R) -2-tert-butoxycarbonylamino-3-phenylpropionylamino) -4-methylpentanamide (354-5)
After 1-1 (148mg, 0.19mmol), HATU (87mg, 0.23mmol) and DIEA (62mg, 0.48mmol) were dissolved in tetrahydrofuran (6 mL), the reaction mixture was stirred at room temperature for 30 minutes, 354-3 (82mg, 0.20mmol) was added to the reaction mixture, the reaction mixture was stirred at room temperature overnight, TLC was performed to monitor the completion of the reaction, DCM (20 mL) was added to the reaction mixture, and 1NNa was used 2 CO 3 Washed separately (10 mL) and 1N HCl (10 mL), and the organic phase was dried and spin dried to give the crude product. Purifying the crude product by column chromatography, eluting with a polar solvent V DCM ：V MeOH =100:1 to 30:1 white solid 354-5 (115 mg, yield: 58.5%) was obtained, ESI-MS (m/z): 1001.4[ M ] +H] + 。
(4) Synthesis of (R) -N- ((R) -1- (2- ((R) -1-acetylpyrrolidine-2-carbonyl) -2, 7-diazaspiro [3.5] nonan-7-yl) -6-amino-1-oxohex-2-yl) -2- ((R) -2- ((R) -2-amino-3-phenylpropanamido) -4-methylpentanamid ditrifluoroacetate (354)
354-5 (115mg, 0.11mmol) was dissolved in methylene chloride (4.5 mL), and trifluoroacetic acid (1.5 mL) was added to the solution, followed by stirring the reaction solution at room temperature for 3 hours. TLC monitoring most of raw materials are consumed, the reaction is stopped, the reaction solution is pulped by ether (10 mL) after being dried, and the solid is filtered and dried to obtain the product. Target compound 354 was a white solid (83 mg, yield: 70.2%), ESI-MS (m/z): 801.3[ 2 ] M + H] + 。
1 HNMR(400MHz,CD 3 OD),δ7.35-7.20(m,10H),4.70(m,1H),4.43-3.92(m,5H),3.82-3.35(m,8H),3.28-3.17(m,2H),3.01-2.90(m,4H),2.21-1.38(m,21H)，0.97(d,3H),0.93(d,3H).
Biological test example
Agonistic activity at the kappa opioid receptor
ForsKolin (ForsKolin) can stimulate the release of human kappa opioid receptor high-expression cell line-OPRKI cell (DiscoveviRx) cAMP, and kappa opioid receptor agonist can inhibit ForsKolin-stimulated cAMP release. The agonistic activity of a compound at the human kappa opioid receptor can be determined by assaying the inhibitory effect of the test compound on forsKolin-stimulated cAMP release. First, a concentration of forsKolin and various concentrations of the test compound were incubated with human high expressing kappa opioid receptor cell line. cAMP immunoassay (LANCEPerKinElmer) based on time-resolved fluorescence resonance energy transfer (TR-FRET) was used to determine cAMP levels in the excited OPRK1 cells. The specific method comprises the following steps:
detection buffer solution: 1 × simulation buffer,500 μ M IBMX, ddH 2 And O. Compound preparation: dissolving the compound with DMSO to prepare a mother solution with a final concentration of 10mM, diluting the mother solution to a working concentration of 2mM, performing 4-fold gradient dilution on the compound with Echo, wherein the initial concentration is 2mM and 10 concentration gradients are added, 50nL is added into a 384 cell plate, the wells are doubled, the final concentration is 10 mu M, and then the cell plate is centrifuged at 1000rpm for 1min; transfer 50nLForsKolin (final concentration 3. Mu.M) to 384 cell plates using Echo.
Cell plating: thawing frozen cells, centrifuging for 5min at 1000rpm, removing supernatant, washing the cells with HBSS buffer solution twice, resuspending the cells with detection buffer solution, and adjusting cell density to 3.0 × 10 5 one/mL, added to 384 well plates at 10 μ L per well, 3000 cells. Shaking for 20s, centrifuging at 1000rpm for 1min, and culturing the cell plate in incubator at 23 deg.C for 60min. Preparation of a standard curve: the standard cAMP was diluted in detection buffer in 4-fold gradient for 8 concentration points with a maximum concentration of 800nM, and 10. Mu.L was added per well according to the microplate layout. Preparing a detection reagent: anti cAMP-Cryptate and AMP-d2 were diluted to 1X with lysis buffer, 10. Mu.L of detection reagent was added to each well according to the layout of the microplate, shaking 20s, centrifuging at 1000rpm for 1min, and the cell plates were incubated in an incubator at 23 ℃ for 60min. Finally the plate was read on Envision.
Percent activity was calculated using Microsoft Excel software, using the formula% Effect = 100X for agonist (Sample Raw Value-Low Control Average)/(High Control Average-Low Control Average). The agonist was selected as Dose-response-Stimulation-log [ aginst ] using GraphPad Prism 5 data analysis software]vs.response--Variable sloCarrying out fitting analysis on the pe mode to obtain the EC of each detection sample 50 The value is obtained. The control group 1 (CR-845, patent CN 1015336A for preparation method reference) and the control group 2 (patent WO2019015644A1 for preparation method reference) have the following structures, respectively, and the experimental data are shown in table 1.
Table 1: agonistic activity of test compounds at kappa opioid receptors
| Compound numbering | EC 50 (nM) |
| 167 | 0.005811 |
| 168 | 0.006291 |
| 176 | 0.04229 |
| 198 | 0.2411 |
| 215 | 0.05256 |
| 229 | 0.008215 |
| 230 | 0.3546 |
| 239 | 0.04643 |
| 256 | 0.001839 |
| 265 | 0.02646 |
| 337 | 0.4266 |
| 343 | 0.464 |
| CR845 | 0.06232 |
| Control group 2 | 0.01154 |
And (4) conclusion: by the above EC 50 As a result, compounds of the embodiments of the present invention have excellent agonistic potency at the kappa opioid receptor. Other compounds of the invention have similarly superior agonistic potency at the kappa opioid receptor.
Rat PK experiment
The compounds were administered intravenously in a single dose (vehicle 5% DMSO +10%Solutol (HS-15) + 85%) to male SD rats (dose 1mg/Kg, 6 per group) and blood was drawn at fixed time points. Immediately after blood collection, the tubes were gently inverted at least 5 times to ensure mixing and then placed on ice. Blood was anticoagulated with heparin and then centrifuged at 8000rpm for 5 minutes to separate serum from erythrocytes. The serum was transferred to a 2mL polypropylene tube using a pipette, the name and time point of the compound was indicated, and stored in a-80 ℃ refrigerator before LC-MS analysis to be tested. After sample treatment, the plasma was analyzed quantitatively by LCMS/MS. Pharmacokinetic parameters were calculated from plasma concentration/time curves obtained in this manner by a validated pharmacokinetic computer program, and the experimental data are shown in table 2:
table 2: results of test Compounds on rat PK
| Compound numbering | Mode of administration | (T 1/2 ,h) | AUClast(h*ng/mL) |
| 265 | iv | 0.22 | 420.9 |
| 256 | iv | 0.14 | 392.7 |
| 168 | iv | 0.2 | 194.4 |
And (4) conclusion: the compound of the invention has better pharmacokinetic property.
All possible combinations of the technical features of the above embodiments may not be described for the sake of brevity, but should be considered as within the scope of the present disclosure as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A compound having a structure represented by formula (II), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:
X 1 and X 2 In, one is CH 2 One is NR, wherein R is
A is
R 1 And R 2 Each independently is: H. c 1-6 Alkyl, - (CH) 2 ) n1 -O-[(CH 2 ) n2 -O] n3 -R a Or- (CH) 2 ) n4 -NR b R c ；
n1, n2, n3, n4 are each independently 1,2,3,4 or 5;
Y 1 is a single bond, CR 11 R 12 、O、S、-(CR 11 ＝CR 12 ) -or-NH-;
R 11 and R 12 Each independently selected from: H. c 1-4 Alkyl, phenyl, 5-6 membered heteroaryl or halogen; said C is 1-4 Alkyl, phenyl, 5-6 membered heteroaryl optionally further substituted with; halogen, cyano, hydroxy, phenyl, -NR b R c 、-COR d 、-COOR d or-CONR b R c ；
B is selected from: 3-10 membered cycloalkyl, 3-10 membered heterocyclyl, 5-10 membered aryl or 5-10 membered heteroaryl;
R 0 is a substituent on B, m1 is 0, 1,2,3,4, 5 or 6;
R 0 at each occurrence, is independently selected from: c 1-4 Alkyl radical, C 1-4 Alkoxy, halogen, cyano, hydroxy, phenyl, 5-6 membered heteroaryl, = O, -NR b R c 、-COR d 、-COOR d 、-CONR b R c or-NHCOR d (ii) a Said C is 1-4 Alkyl radical, C 1-4 Alkoxy, phenyl, 5-6 membered heteroaryl optionally further substituted with: halogen, cyano, hydroxy or phenyl;
R a is C 1-4 An alkyl group;
R b 、R c each independently selected from: H. c 1-4 Alkyl or C 1-4 An alkoxy group;
R d selected from: h or C 1-4 An alkyl group.
2. The compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, wherein B is selected from: 3-8 membered monocyclic alkyl, 5-6 membered cycloalkylacenyl, 5-6 membered heterocyclo-5-6 membered heterocyclyl, 5-6 membered cycloalkylo-5-6 membered heterocyclyl, 5-6 membered heterocyclo-phenyl, 5-6 membered heterocyclo-5-6 membered heteroaryl, 6-10 membered aryl, or 5-10 membered heteroaryl.
3. The compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, which is selected from compounds represented by any one of the following general formulae:
B 1 selected from: 3-8 membered monocycloalkyl, 5-6 membered cycloalkylacenyl, 5-6 membered heteromonocyclic group, 5-6 membered heterocyclo-5-6 membered heterocyclic group, 5-6 membered cycloalkylaceno 5-6 membered heterocyclic group, 5-6 membered heterocyclo-phenyl, 6-10 membered aryl, or 5-10 membered heteroaryl;
B 2 selected from: 6-10 membered aryl or 5-10 membered heteroaryl;
B 3 selected from: 6-10 membered aryl or 5-10 membered heteroaryl;
B 4 selected from: 6-10 membered aryl or 5-10 membered heteroaryl.
4. A compound, pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 2 or 3, wherein the 3-8 membered monocycloalkyl is selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, or cyclooctyl;
5-6 membered mono heterocyclic group selected from: aziridinyl, oxiranyl, azetidinyl, oxetanyl, oxazolidinyl, 1, 3-dioxolanyl, dioxanyl, imidazolidinyl, tetrahydrofuryl, tetrahydrothienyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, thiomorpholine-1, 1-dioxide, tetrahydropyranyl, 1, 3-oxazinoalkyl, hexahydropyrimidyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, 1, 3-oxazepanyl, 1, 2-dihydroazetidinyl, 2, 5-dihydro-1H-pyrrolyl, 2, 3-dihydro-1H-pyrrolyl, 1,2,3, 4-tetrahydropyridinyl, 1,2,3, 6-tetrahydropyridinyl, 2-pyrrolidinonyl or 2-piperidinonyl;
5-6 membered heterocyclyl and 5-6 membered heterocyclyl is selected from any one of the following:
the 5-6 membered cycloalkyl and 5-6 membered heterocyclyl is selected from any one of the following:
the 5-6 membered cycloalkylacenyl is selected from: a cyclopentylphenyl group, or a cyclohexylphenyl group;
5-6 membered heterocyclo-phenyl any of the following:
6-10 membered aryl is selected from: benzene or naphthalene;
the 5-10 membered heteroaryl is selected from: thiophene, furan, thiazole, isothiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, 1,2, 3-triazole, 1,2, 4-triazole, 1,2, 5-triazole, 1,3, 4-triazole, tetrazole, isoxazole, oxadiazole, 1,2, 3-oxadiazole, 1,2, 4-oxadiazole, 1,2, 5-oxadiazole, 1,3, 4-oxadiazole, thiadiazole, pyridine, pyridazine, pyrimidine, pyrazine or any of the following groups:
5. the compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 1, wherein R is 0 At each occurrence, each is independently selected from: c 1-4 Alkyl, halogen, -OCF 3 、-NH 2 、-COOR d or-CONR b R c 。
6. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 1,
selected from any one of the following:
z is NH, O, S, CR h R j Or C = O; w is CR h Or N; v is CR h Or N; and two adjacent Z are not hetero atoms at the same time;
R h and R j Each independently selected from: H. c 1-4 Alkyl radical, C 1-4 Alkoxy, halogen, cyano, hydroxy, phenyl, 5-6 membered heteroaryl, -NR b R c 、-COR d 、-COOR d 、-CONR b R c or-NHCOR d (ii) a Said C is 1-4 Alkyl radical, C 1-4 Alkoxy, phenyl, 5-6 membered heteroaryl optionally further substituted with: halogen, cyano, hydroxy, phenyl, -NR b R c or-COOR d 。
7. The compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 1,
selected from any one of the following structures:
8. a compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 1, characterized by-NR 1 R 2 Selected from the group consisting of: -NH 2 、-NH-(CH 2 ) 2 -O-(CH 2 ) 2 -O-R a 、-NH(CH 2 ) 3 -NR b R c or-NH (CH) 2 ) 4 -NR b R c ；R a Is C 1-3 An alkyl group; r b 、R c Each independently selected from: h or C 1-3 An alkyl group.
9. The compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 1, which is selected from any one of the following compounds:
10. use of a compound, pharmaceutically acceptable salt, stereoisomer, or co-crystal thereof of any one of claims 1 to 9 in the manufacture of a medicament for treating or preventing a disease or condition associated with a kappa opioid receptor selected from pain, inflammation, itch, edema, hyponatremia, hypokalemia, ileus, cough, and glaucoma.
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