CN118255685A - Tricyclic compound, pharmaceutical composition and application thereof - Google Patents

Tricyclic compound, pharmaceutical composition and application thereof Download PDF

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Publication number
CN118255685A
CN118255685A CN202311773333.3A CN202311773333A CN118255685A CN 118255685 A CN118255685 A CN 118255685A CN 202311773333 A CN202311773333 A CN 202311773333A CN 118255685 A CN118255685 A CN 118255685A
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alkyl
independently
membered
substituted
compound
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刘金镖
唐静洁
邓向君
高昂
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Kelan Shanghai Pharmaceutical Technology Co ltd
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Kelan Shanghai Pharmaceutical Technology Co ltd
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Abstract

The invention discloses a tricyclic compound, a pharmaceutical composition and application thereof. Specifically, the invention discloses a compound shown as a formula (I) or pharmaceutically acceptable salt thereof. The compound shown as the formula (I) or the pharmaceutically acceptable salt thereof has one or more of the following advantages: the SMO inhibitor has good SMO inhibition; shows good Hh signal path inhibition ability; has good metabolism stability of liver microsome.

Description

Tricyclic compound, pharmaceutical composition and application thereof
Technical Field
The invention relates to a tricyclic compound, a pharmaceutical composition thereof and application thereof.
Background
Hedgehog (Hh) signaling pathway plays an important role in embryogenesis in organogenesis and is involved in tissue repair and homeostasis maintenance in adults. The Hh signaling pathway mainly includes secreted ligands (SHH, IHH and DHH), the negative regulatory receptor Patched (PTCH), the positive regulatory protein Smoothened (SMO), and GLI transcription factors.
Abnormal activation of the Hh pathway has been found in a variety of tumors. 90% of patients with basal cell carcinoma (basal cell carcinomas) and about 20% of patients with medulloblastoma (medulloblastomas) have constitutive activation of the Hh pathway by PTCH inactivating mutations or acquired mutation of SMO. In addition, pathway activation due to Hh ligand overexpression is also present in a variety of tumors, including pancreatic cancer, colon cancer, gastric cancer, lung cancer, breast cancer, leukemia, multiple myeloma, and the like.
SMO is a 7-transmembrane G-protein coupled receptor-like protein. Three small molecule inhibitors developed for SMO are currently commercially available. Including vmod gide (Vismodegib) and sonid gide (Sonidegib) against basal cell carcinoma, and glad gide (Glasdegib) against leukemia. Binding sites for all three drugs are located in the transmembrane region (TMD) of SMO. The better response rates of vemod and sony dyad against basal cell carcinoma, but one important issue is the rapid development of drug resistance mutations following treatment. Wherein about half of the drug resistance mutations occur in the transmembrane region of SMO, about 17% of the mutations are D473 mutations.
The extracellular cysteine rich region (CRD) of SMO can be activated by covalent modification with a steroid ligand, and drugs targeting the CRD region are expected to inhibit drug-resistant mutations and to act synergistically with drugs already on the market for TMD.
Disclosure of Invention
The invention aims to overcome the defect of lack of inhibitors targeting SMO in the prior art, and provides a tricyclic compound with a novel structure, a preparation method thereof, a pharmaceutical composition thereof and application thereof. The tricyclic compounds of the invention have one or more of the following advantages: the SMO inhibitor has good SMO inhibition; shows good Hh signal path inhibition ability; has good metabolism stability of liver microsome.
The present invention solves the above-mentioned problems by the following method.
The invention provides a compound shown as a formula (I) or pharmaceutically acceptable salt thereof:
Wherein,
R 1 is-OH, -NH 2 or-NR 1-1SO2R1-2; or R 1 is bound to a carbon atom to form
R 1-1 and R 1-2 are independently H or C 1-C6 alkyl;
L 1 is- (CH 2) n-where n is an integer from 1 to 12, 0,1, 2 or 3-CH 2 -are replaced by-CHR L-1 -or-CR L- 1RL-1 -;
R L-1 is independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-C(＝O)NR2-1-、-NR2-1C(＝O)-、-NR2-1C(＝O)NR2-1-、-C(＝O)-、-C(＝O)O-、-C(＝O)NR2-1NR2-1 C(＝O)-、-NR2-1C(＝O)O-、-SO2NR2-1- or 5-10 membered heteroarylene, in L 2, the hetero atom in the heteroarylene is one, two or three selected from N, O and S, and the number of the hetero atoms is 1, 2 or 3;
r L-2' is OH; r L-2 is H or C 1-C6 alkyl;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1;
R 2-1-1 is independently OH, COOH or C 1-C6 alkoxy;
L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, -CHR L-3-、-NRL-4 -, -O-, -S-or 3-6 membered heterocycloalkylene; in L 3, in the heterocycloalkylene group, the type of the heteroatom is one, two or three selected from N, O and S, and the number of the heteroatom is 1, 2 or 3;
R L-3 is independently C 1-C6 alkyl or C 1-C6 alkoxy;
R L-4 is independently H or C 1-C6 alkyl, C 1-C6 alkoxy, C 1-C6 alkyl substituted with 1, 2 or 3R 2-2;
R 2-2 is OH, NR 2-2-1R2-2-1, halogen, C 1-C6 alkoxy, 5-6 membered heteroaryl, -C (=O) NR 2-2-1R2-2-1、-NR2 -2-1C(＝O)-C1-C6 alkyl, -NR 2-2-1C(＝O)NR2-2-1R2-2-1、-C(＝O)-C1-C6 alkyl, -C (=O) OR 2-2-1、-C(＝O)NR2-2-1NR2-2-1C(＝O)C1-C6 alkyl OR-SO 2NR2-2-1R2-2-1;R2-2, wherein the hetero atoms are selected from one, two OR three of N, O and S, and the number of the hetero atoms is 1, 2 OR 3;
R 2-2-1 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4, wherein in A, the type of heteroatom is selected from one, two or three of N, O and S, and the number of heteroatoms is 1, 2, 3 or 4; in the A, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1, 2 or 3;
R 3-1 is independently oxo, CN, NR 3-1-1R3-1-2、COOR3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1, 2 or 3R 3-1-4, wherein in the heterocycloalkyl, the type of heteroatom is selected from one, two or three of N, O and S, and the number of the heteroatom is 1, 2 or 3;
r 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl;
R 3-1-4 is independently halogen or 5-10 membered heteroaryl, wherein in the heteroaryl, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2, 3 or 4;
R 3-2 is independently halogen, (c=o) NR 3-2-3R3-2-4、C1-C6 alkyl, C 1-C6 alkoxy, -NR 3-2-1(S＝O)2R3 -2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
wherein the compound represented by formula (I) is not the following compound or stereoisomer thereof:
The invention provides a compound shown as a formula (I) or pharmaceutically acceptable salt thereof:
Wherein,
R 1 is-OH, -NH 2 or-NR 1-1SO2R1-2; or R 1 is bound to a carbon atom to form
R 1-1 and R 1-2 are independently H or C 1-C6 alkyl;
L 1 is- (CH 2) n-where n is an integer from 1 to 12, 0,1, 2 or 3-CH 2 -are replaced by-CHR L-1 -or-CR L- 1RL-1 -;
R L-1 is independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-C(＝O)NR2-1-、-NR2-1C(＝O)-、-NR2-1C(＝O)NR2-1-、-C(＝O)-、-C(＝O)O-、-C(＝O)NR2-1NR2-1C(＝O)-、-NR2-1C(＝O)O-、-SO2NR2-1- or 5-6 membered heteroarylene, in L 2, the hetero atom in the heteroarylene is one, two or three selected from N, O and S, and the number of the hetero atoms is 1,2 or 3;
r L-2' is OH; r L-2 is H or C 1-C6 alkyl;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1;
R 2-1-1 is independently OH, COOH or C 1-C6 alkoxy;
L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, -CHR L-3-、-NRL-4 -, -O-, -S-or 3-6 membered heterocycloalkylene; in L 3, in the heterocycloalkylene group, the type of the heteroatom is one, two or three selected from N, O and S, and the number of the heteroatom is 1, 2 or 3;
R L-3 is independently C 1-C6 alkyl or C 1-C6 alkoxy;
R L-4 is independently H or C 1-C6 alkyl, C 1-C6 alkoxy, C 1-C6 alkyl substituted with 1, 2 or 3R 2-2;
R 2-2 is OH, NR 2-2-1R2-2-1, halogen, C 1-C6 alkoxy, 5-6 membered heteroaryl, -C (=O) NR 2-2-1R2-2-1、-NR2 -2-1C(＝O)-C1-C6 alkyl, -NR 2-2-1C(＝O)NR2-2-1R2-2-1、-C(＝O)-C1-C6 alkyl, -C (=O) OR 2-2-1、-C(＝O)NR2-2-1NR2-2-1C(＝O)C1-C6 alkyl OR-SO 2NR2-2-1R2-2-1;R2-2, wherein the hetero atoms are selected from one, two OR three of N, O and S, and the number of the hetero atoms is 1, 2 OR 3;
R 2-2-1 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4, wherein in A, the type of heteroatom is selected from one, two or three of N, O and S, and the number of heteroatoms is 1, 2, 3 or 4; in the A, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1, 2 or 3;
R 3-1 is independently CN, NR 3-1-1R3-1-2、COOR3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1,2 or 3R 3-1-4, wherein the hetero atoms are selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2 or 3;
r 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl;
R 3-1-4 is independently halogen or 5-10 membered heteroaryl, wherein in the heteroaryl, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2, 3 or 4;
R 3-2 is independently halogen, (c=o) NR 3-2-3R3-2-4、C1-C6 alkyl, C 1-C6 alkoxy, -NR 3-2-1(S＝O)2R3 -2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Wherein the compound shown in the formula (I) is not the following compound:
In certain preferred embodiments of the present invention, certain groups in the compounds of formula (I) or pharmaceutically acceptable salts thereof are defined below, and the unrecited groups are as recited in any of the embodiments of the present invention (referred to herein as "in one embodiment").
In one embodiment of the invention ,R1-1、R1-2、RL-1、RL-2、R2-1、RL-3、RL-4、R2-2、R3-1、R3-3、R2-2-1、R3 -1-1、R3-1-2、R3-1-3、R3-2、R3-2-1、R3-2-2、R3-2-3、R3-2-4、 and R 3-4, each "C 1-C6 alkyl" may independently be methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, preferably methyl, ethyl or tert-butyl.
In one embodiment of the invention, when L 2 is a 5-10 membered heteroarylene, the 5-10 membered heteroarylene may be a 5-6 membered monocyclic heteroarylene or a 9-10 membered bicyclic heteroarylene, for example, a 5-6 membered heteroarylene; the hetero atom in the heteroarylene group is selected from one or both of N and O, and the number of hetero atoms is 2 or 3, for example, triazolylene, oxadiazolylene or benzimidazolylene, further for example, 1,2, 3-triazolylene, 1,3, 4-oxadiazolylene, 1,2, 4-oxadiazolylene or benzimidazolylene, preferably Further preferred is Wherein the j side is connected with L 1.
In one embodiment of the present invention, when L 2 -6 membered heteroarylene is selected from the group consisting of N and O, the 5-6 membered heteroarylene may be a5 membered heteroarylene having 2 or 3 hetero atoms, such as triazolylene or oxadiazolylene, further such as 1,2, 3-triazolylene, 1,3, 4-oxadiazolylene or 1,2, 4-oxadiazolylene, preferably Further preferred isWherein the j side is connected with L 1.
In one embodiment of the invention, R 2-1-1、RL-3、RL-4、R2-2 and R 3-2, the C 1-C6 alkoxy group may independently be methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy.
In one embodiment of the present invention, the 3-6 membered heterocycloalkylene group in L 3 may be one or two of N and O, and the 5-6 membered heterocycloalkylene group having 1 or 2 hetero atoms, such as piperidylene, tetrahydropyranyl, piperazinylene or morpholinylene, further such as piperidylene, preferably
In one embodiment of the invention, in A, each "C 6-C10 aryl" may independently be phenyl or naphthyl.
In one embodiment of the invention, each "5-10 membered heteroaryl" in A may independently be a 5-6 membered heteroaryl or a 9-10 membered bicyclic heteroaryl, such as pyridinyl, oxadiazolyl, azaindolyl, benzimidazolyl, indolyl, benzothiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, indazolyl, benzotriazolyl, indolinyl, benzomorpholinyl, benzoxadiazolyl, quinazolinyl, quinolinyl, isoquinolinyl, 4,5,6, 7-tetrahydro-1H-benzimidazolyl or 2, 3-dihydrobenzimidazolyl, for example
In one embodiment of the invention, each "5-10 membered heteroaryl" is independently a 5-6 membered heteroaryl or a 9-10 membered bicyclic heteroaryl, such as pyridinyl, oxadiazolyl, azaindolyl, benzimidazolyl, indolyl, benzothiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, indazolyl, benzotriazolyl, indolinyl, benzomorpholinyl, benzoxadiazolyl, quinazolinyl, quinolinyl, or isoquinolinyl, more such as
In one embodiment of the present invention, R 2-2 -6 membered heteroaryl may be one or two of N and O, and the number of the heteroatoms is 1 or 2, for example, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyridazinyl or pyrazinyl.
In one embodiment of the invention, each of A and R 3-1, each "C 3-C6 cycloalkyl" may independently be cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
In one embodiment of the invention, each of A and R 3-1, 3-6 membered heterocycloalkyl may independently be a heteroatom selected from one or both of N and O, a 5-6 membered heterocycloalkyl having 1 or 2 heteroatoms, such as piperidinyl, tetrahydropyranyl, morpholinyl or piperazinyl, further such as
In one embodiment of the invention, each "halogen" in R 2-2、R3-1、R3-1-4 and R 3-2 may independently be F, cl, br or I, such as F or Cl.
In one embodiment of the invention, R 1 is-OH or-NR 1-1SO2R1-2.
In one embodiment of the invention, R 1-1 is H.
In one embodiment of the invention, R 1-2 is C 1-C6 alkyl.
In one embodiment of the invention, R 1 is-OH or-NR 1-1SO2R1-2,R1-1 is H and R 1-2 is C 1-C6 alkyl. Preferably, R 1 is-OH or-NHSO 2CH3.
In one embodiment of the invention, 0, 1, 2 or 3-CH 2 -in L 1 are replaced with CHR L-1.
In one embodiment of the invention, n is 2, 3 or 4, preferably 2 or 3.
In one embodiment of the invention, L 1 is- (CH 2) n-where n is an integer from 1 to 12, where 0, 1, 2 or 3-CH 2 -are replaced by CHR L-1;RL-1 is C 1-C6 alkyl; preferably, n is 2, 3 or 4; more preferably, n is 2 or 3.
In one embodiment of the invention, L 1 is Preferably is Wherein side c is connected to L 2.
In one embodiment of the invention, R 2-1 is H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3C 1-C6 alkoxy groups.
In one embodiment of the invention, L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NR2-1NR2-1(C＝O)-、-NR2-1(C＝O)O- or a 5-to 10-membered heteroarylene, preferably -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NR2-1NR2-1(C＝O)-、-NR2-1(C＝O)O- or a 5-to 6-membered heteroarylene; preferably, L 2 is -CH(OH)-、-NH-、-(C＝O)NR2-1-、-NH(C＝O)-、-NH(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NHNH(C＝O)-、-NH(C＝O)O- or a 5-6 membered heteroarylene; more preferably, L 2 is- (c=o) NR 2-1、-NH(C＝O)-、-NH(C＝O)NH-、-NH-,R2-1 is H, C 1-C6 alkyl or C 1-C6 alkyl substituted by 1,2 or 3C 1-C6 alkoxy groups.
In one embodiment of the invention, L 2 is
Preferably is
Wherein the d-side is connected to L 1.
In one embodiment of the invention, L 2 is
Preferably is
Wherein the d-side is connected to L 1.
In one embodiment of the invention, 0, 1,2 or 3-CH 2 -in L 3 are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene.
In one embodiment of the invention, m is 1,2, 3, 4, 5, 6, 7, 8, 9, 10 or 11.
In one embodiment of the invention, R L-4 is H or C 1-C6 alkyl.
In one embodiment of the present invention, when R L-4 is C 1-C6 alkyl substituted with 1,2 or 3R 2-2, R 2-2 is at the end of C 1-C6 alkyl, preferably R 2-2 is 1.
In one embodiment of the invention, L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1,2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is H or C 1-C6 alkyl; preferably, L 3 is a bond or- (CH 2) m-, m is 1,2, 3, 4, 5,6, 7, 8, 9, 10 or 11, wherein 0 or 1-CH 2 -in- (CH 2) m-is replaced by-CH=CH-, NH, O or 3-6 membered heterocycloalkylene, when 0 or 1-CH 2 -in- (CH 2) m-is replaced with a 3-6 membered heterocycloalkylene group, the 3-6 membered heterocycloalkylene group is linked to L 2.
In one embodiment of the invention, L 3 is a bond,
Preferably a connecting bond,
Wherein the e side is connected with A.
In one aspect of the invention, -L 1-L2-L3 -is any one of the following:
i: -in L 1-L2-L3 -, L 1 is- (CH 2) n-, n is 2,3 or 4, wherein 0 or 1 of- (CH 2) n-CH 2 -are replaced by CHR L-1 -or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-NR2-1(C＝O)O- or 5-6 membered subunit
Heteroaryl; r L-2' is OH; r L-2 is H or C 1-C6 alkyl; r 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 2-1-1; r 2-1-1 is independently COOH or C 1-C6 alkoxy;
L 3 is- (CH 2) m-, m is an integer from 1 to 12, wherein, 0,1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is independently H or C 1-C6 alkyl;
ii: -in L 1-L2-L3 -, L 1 is- (CH 2) n-, n is 2,3 or 4, wherein 0, 1, 2 or 3-CH 2 -are replaced by CHR L -1 or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-to 10-membered heteroarylene, preferably -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)- or 5-to 6-membered heteroarylene;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
l 3 is a connecting bond.
In one embodiment of the invention, -L 1-L2-L3 -is
Preferably is
Wherein the f side is connected with A.
In one embodiment of the invention R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 3-1-4, preferably CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 3-1-4.
In one embodiment of the invention, R 3-1-1 and R 3-1-2 are independently H.
In one embodiment of the invention, R 3-1-3 is H or C 1-C6 alkyl.
In one embodiment of the invention, R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, or C 1-C6 alkyl substituted with 1,2, or 3R 3-1-4, R 3-1-1 and R 3-1-2 are independently H, R 3-1-3 is H or C 1-C6 alkyl, and R 3-1-4 is independently halogen or 5-10 membered heteroaryl.
In one embodiment of the invention, R 3-1 is independently CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4, R 3-1-1 and R 3-1-2 are independently H, and R 3-1-3 is H or C 1-C6 alkyl.
In one embodiment of the invention, R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3 -2-2 or NR 3-2-3R3-2-4.
In one embodiment of the invention, R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl.
In one embodiment of the invention, R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3 -2-2 or NR 3-2-3R3-2-4,R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 is independently C 1-C6 alkyl.
In one embodiment of the invention, A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2, or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3-4;
R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4, R 3-1-4 is independently halogen or 5-10 membered heteroaryl; preferably CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1,2 or 3R 3-1-4;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Preferably, A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1 or C 6-C10 aryl substituted with 1,2 or 3R 3-2;
R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4, R 3-1-4 is independently halogen or 5-10 membered heteroaryl; preferably CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1,2 or 3R 3-1-4;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl.
In one embodiment of the invention, A is-NR 3-3R3-3、C6-C10 aryl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3 -4;
R 3-1 is independently oxo, CN, COOR 3-1-3、C1-C6 alkyl, halogen, CONR 3-1-1R3-1-2、C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
R 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl; r 3-1-4 is independently halogen or 5-10 membered heteroaryl;
r 3-2 is independently OH, NR 3-2-1(S＝O)2R3-2-2、NR3-2-3R3-2-4;R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
r 3-4 is independently C 1-C6 alkyl.
In one embodiment of the invention, A is H,
Preferably H,
In one embodiment of the invention, -L 1-L2-L3 -A is any one of the following:
I: -in L 1-L2-L3 -a, L 1 is- (CH 2) n-, n is 2, 3 or 4, wherein 0 or 1 of- (CH 2) n-CH 2 -are replaced by CHR L-1 -or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-NR2-1(C＝O)O- or 5-6 membered subunit
Heteroaryl; r L-2' is OH; r L-2 is H or C 1-C6 alkyl; r 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 2-1-1; r 2-1-1 is independently COOH or C 1-C6 alkoxy;
L 3 is- (CH 2) m-, m is an integer from 1 to 12, wherein, 0,1, 2 or 3-CH 2 -in- (CH 2) n-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-C10 aryl substituted with 1, 2 or 3R 3-2, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4;
R 3-1 is C 1-C6 alkyl; r 3-2 is independently halogen, C 1-C6 alkyl or (c=o) NR 3-2-3R3-2-4;R3-2-3 and R 3-2-4 is independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
II: -in L 1-L2-L3 -a, L 1 is- (CH 2) n-, n is 2,3 or 4, wherein 0,1, 2 or 3-CH 2 -are replaced by CHR L-1 or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-to 10-membered heteroarylene group;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
L 3 is a bond;
A is-NR 3-3R3-3、C6-C10 aryl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3-4;
R 3-1 is independently oxo, CN, COOR 3-1-3、C1-C6 alkyl, halogen, CONR 3-1-1R3-1-2、C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
R 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl; r 3-1-4 is independently halogen or 5-10 membered heteroaryl;
r 3-2 is independently OH, NR 3-2-1(S＝O)2R3-2-2、NR3-2-3R3-2-4;R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Preferably, in-L 1-L2-L3 -A, L 1 is- (CH 2) n-, n is 2, 3 or 4, wherein 0, 1, 2 or 3-CH 2 -are replaced by CHR L-1 or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-6 membered heteroarylene group;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
L 3 is a bond;
A is C 6-C10 aryl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3-4;
R 3-1 is CN, COOR 3-1-3、C1-C6 alkyl, halogen, CONR 3-1-1R3-1-2、C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 3-1-4;
R 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl; r 3-1-4 is independently halogen or 5-10 membered heteroaryl;
r 3-2 is independently OH, NR 3-2-1(S＝O)2R3-2-2、NR3-2-3R3-2-4;R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
r 3-4 is independently C 1-C6 alkyl.
In one embodiment of the invention, -L 1-L2-L3 -A is:
Preferably is
In one embodiment of the present invention, the compound represented by formula (I) is a compound represented by formula (I-A):
Wherein R 1、L1、L2、L3 and A are as defined in any one of the embodiments of the present invention.
In one embodiment of the present invention, the compound represented by formula (I) is a compound represented by formula (I-B):
Wherein,
R 1 is-OH or-NR 1-1SO2R1-2;R1-1 and R 1-2 is independently H or C 1-C6 alkyl;
L 2 is-NR L-2-、-C(＝O)NR2-1-、-NR2-1 C (=o) -or-NR 2-1C(＝O)NR2-1-,R2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted by 1,2 or 3R 2-1-1; r 2-1-1 is independently C 1-C6 alkoxy; r L-2 is H or C 1-C6 alkyl;
L 3 is a bond or- (CH 2) m-, m is 1, 5, 6, 7 or 8, 0 or 1 of the- (CH 2) m-s-CH 2 -s are replaced by-ch=ch-, -NH-, or-O-;
A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2;
R 3-1 and R 3-2 are as defined in any one of the embodiments of the invention.
In one embodiment of the present invention, the compound represented by formula (I) is a compound represented by formula (I-C):
Wherein, Is 3-6 membered heterocycloalkylene; q is- (CH 2)2-4 -; A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1 or C 6-C10 aryl substituted with 1,2 or 3R 3-2, R 3-1 and R 3-2 are as defined in any of the embodiments of the invention.
In one embodiment of the present invention, the compound represented by formula (I) is a compound represented by formula (I-D):
Wherein L 2 is-NR 2-1C(＝O)N R2-1-、-C(＝O)NR2-1 -or-NR 2-1-C(＝O)-,R2-1 is H or C 1-C6 alkyl;
L 3 is- (CH 2) m-and m is 9,10 or 11;
A is H, CN, -NR 3-3R3-3 OR-OR 3-3;
R 3-3 is independently H or C 1-C6 alkyl.
The present invention also provides a compound as shown below or a pharmaceutically acceptable salt thereof:
In one embodiment of the present invention, the pharmaceutically acceptable salts of the compounds of formula (I) are:
The invention also provides a pharmaceutical composition which comprises a substance X and a pharmaceutically acceptable carrier, wherein the substance X is a compound shown in the formula (I) or pharmaceutically acceptable salt thereof.
The invention also provides an application of the compound shown in the formula (I), the pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing an SMO inhibitor.
The invention also provides an application of the compound shown in the formula (I), the pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing a medicament for treating and/or preventing SMO related diseases.
The invention also provides an application of the compound shown in the formula (I), the pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing medicines for treating and/or preventing diseases related to abnormal activation of Hh signal channels.
The invention also provides an application of the compound shown in the formula (I), pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparing medicines for treating and/or preventing basal cell carcinoma or medulloblastoma and other diseases.
On the basis of conforming to the common knowledge in the field, the above preferred conditions can be arbitrarily combined to obtain the preferred examples of the invention.
The reagents and materials used in the present invention are commercially available.
Unless otherwise indicated, the terms used in the present invention have the following meanings:
Those skilled in the art will appreciate that, in accordance with the convention used in the art, the present invention describes the structural formula of the group used Meaning that the corresponding group is linked to other fragments, groups in the compound through this site.
As used herein, a substituent may be appended with a single dash "-" indicating that the named substituent is attached to the parent moiety through a single bond. It is also possible to add "=", indicating that the named substituents are attached to the parent moiety through a double bond.
If a linking group is denoted as "bond", it means that the structures on both sides of the linking group are directly linked, e.g. -A-B-C-, when B is a bond, -A-B-C-is-A-C-.
The term "pharmaceutically acceptable" refers to salts, solvents, excipients, and the like, which are generally non-toxic, safe, and suitable for patient use. The "patient" is preferably a mammal, more preferably a human.
The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared with relatively non-toxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a pharmaceutically acceptable base in pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to: lithium salt, sodium salt, potassium salt, calcium salt, aluminum salt, magnesium salt, zinc salt, bismuth salt, ammonium salt, diethanolamine salt. When the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in pure solution or in a suitable inert solvent. The pharmaceutically acceptable acids include inorganic acids including, but not limited to: hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, sulfuric acid, and the like. The pharmaceutically acceptable acid includes organic acids including, but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acidic citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentisic acid, fumaric acid, gluconic acid, sugar acid, formic acid, ethanesulfonic acid, pamoic acid (i.e., 4' -methylene-bis (3-hydroxy-2-naphthoic acid)), amino acids (e.g., glutamic acid, arginine), and the like. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. See, for example, berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science 66:1-19 (1977), or 、Handbook of Pharmaceutical Salts:Properties,Selection,and Use(P.Heinrich Stahl and Camille G.Wermuth,ed.,Wiley-VCH,2002).
The terms "compound" and "pharmaceutically acceptable salt" when present, may exist as single stereoisomers or as mixtures thereof (e.g., racemates). The term "stereoisomer" refers to a cis, trans or optical isomer. These stereoisomers may be isolated, purified and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin layer chromatography, rotary chromatography, column chromatography, gas chromatography, high pressure liquid chromatography, etc.), and may be obtained by chiral resolution by bonding (chemical bonding, etc.) or salifying (physical bonding, etc.) other chiral compounds. The term "single stereoisomer" means that one stereoisomer of the compound of the present invention is present in an amount of not less than 95% by mass relative to all stereoisomers of the compound.
When any variable (e.g., R 3-1) occurs multiple times in the definition of a compound, the definition of each position of the variable occurs independently of the definition of the other positions, their meanings are independent of each other and do not affect each other. Thus, if a group is substituted with 1, 2 or 3R 3-1 groups, that is, the group may be substituted with up to 3R 3-1, the definition of position R 3-1 is independent of the definition of the remaining positions R 3-1. In addition, combinations of substituents and/or variables are allowed only if the combination yields a stable compound.
The "inhibitors" of the invention are useful in mammalian organisms; it is also useful in vitro, mainly as an experimental use, for example: the kit can be used as a standard sample or a control sample for comparison or prepared according to a conventional method in the field, and can be used for rapidly detecting the SMO inhibition effect.
The term "alkyl" refers to a straight or branched chain alkyl group having the indicated number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl and the like.
The term "halogen" refers to F, cl, br or I.
The term "cycloalkyl" refers to a saturated monocyclic group having the specified number of ring carbon atoms (e.g., C 3-C6), the ring atoms consisting of only carbon atoms. Including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
The term "alkylene" refers to a saturated, straight-chain, divalent hydrocarbon radical having the indicated number of carbon atoms. Thus, C 1 alkylene (i.e., methylene) refers to-CH 2-,C2 alkylene (i.e., ethylene) refers to-CH 2-CH2-,C3 alkylene refers to-CH 2-CH2-CH2 -.
The term "alkenylene" refers to a straight-chain divalent hydrocarbon radical having the indicated number of carbon atoms and at least one carbon-carbon double bond, where the carbon-carbon double bond may be located anywhere within the alkenylene radical. Thus, C 2 alkenylene (i.e., vinylidene) refers to-ch=ch-, C 3 alkenylene refers to-CH 2 -ch=ch-, and-CH 2＝CH-CH2-,C4 alkenylene refers to-CH 2-CH＝CH-CH2-、-CH2＝CH-CH2-CH2 -, and-CH 2-CH-CH2＝CH2 -.
The term "heteroaryl" refers to a cyclic, unsaturated monovalent group of a specified number of heteroatoms (e.g., 1, 2,3 or 4) having a specified number of ring atoms (e.g., 5-10 members), of a specified heteroatom species (1, 2 or 3 of N, O and S), which is monocyclic or bicyclic, where two atoms and one bond are shared between each monocyclic ring, and at least one or each ring has aromaticity, through which the heteroaryl group may be attached to other moieties in the molecule. Examples of heteroaryl groups include, but are not limited to, oxazolyl, oxadiazolyl, triazolyl, pyridyl, azaindolyl, benzimidazolyl, indolyl, benzothiadiazolyl, benzoxadiazolyl, pyridazinyl, pyrimidinyl, indazolyl, pyrazinyl, benzotriazolyl, indolinyl, benzomorpholinyl, quinolinyl, isoquinolinyl, 4,5,6, 7-tetrahydro-1H-benzimidazolyl, 2, 3-dihydrobenzimidazolyl, and the like.
The term "heteroarylene" refers to a divalent heteroaryl group. Examples of 5-membered heteroarylenes, as defined above, include, but are not limited to oxadiazolyl or triazolyl.
The term "alkoxy" refers to the group-O-R X, wherein R X is alkyl as defined above.
The term "aryl" refers to any stable monocyclic or polycyclic (e.g., monocyclic or bicyclic) carbocycle of up to 6 atoms in each ring, each of which is aromatic.
The term "heterocycloalkyl" refers to a saturated cyclic group of a specified number of heteroatoms (e.g., 1,2, 3, or 4) of a specified number of ring atoms (e.g., 3-6 members), of a specified heteroatom species (1, 2, or 3 of N, O and S), which is a single ring. Heterocycloalkyl groups include, but are not limited to, tetrahydropyranyl, piperazinyl, morpholinyl, piperidinyl, and the like.
The term "pharmaceutically acceptable excipients" refers to excipients and additives used in the manufacture of medicaments and formulation prescriptions, and are all substances contained in the pharmaceutical formulation except the active ingredient. See the pharmacopoeia of the people's republic of China (2015 Edition), or Handbook of Pharmaceutical Excipients (Raymond C Rowe,2009 sibth Edition).
The term "treatment" refers to therapeutic therapy. When specific conditions are involved, treatment refers to: (1) alleviating a disease or one or more biological manifestations of a disorder, (2) interfering with (a) one or more points in a biological cascade that results in or causes a disorder or (b) one or more biological manifestations of a disorder, (3) ameliorating one or more symptoms, effects, or side effects associated with a disorder, or one or more symptoms, effects, or side effects associated with a disorder or treatment thereof, or (4) slowing the progression of a disorder or one or more biological manifestations of a disorder.
The term "preventing" refers to a reduced risk of acquiring or developing a disease or disorder.
The invention has the positive progress effects that: the compounds of the present invention have one or more of the following advantages: the SMO inhibitor has good SMO inhibition; shows good Hh signal path inhibition ability; has good metabolism stability of liver microsome.
Detailed Description
The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.
In the following examples:
Source of NIH3T3/Gli-luc/12 cells
NIH-3T3 cells were purchased from the China academy of sciences typical culture Collection Committee cell Bank, catalogue number: SCSP-515.NIH-3T3/Gli-Luc/12 cells were transfected with pGliBS and pRL-TK-IRES-ZSGREEN plasmids on the basis of NIH-3T3 cells and were screened by puromycin to obtain successfully constructed monoclonal cells. The cell line stably expresses RL-TK and ZSGREEN1, and can express luciferase protein under the stimulation of signals.
Wherein NIH-3T3 cell line, pGliBS plasmid information is described in the following references:
Fu,L.,Wu,H.,Cheng,S.Y.,Gao,D.,Zhang,L.,and Zhao,Y.(2016).Set7 mediated Gli3 methylation plays a positive role in the activation of Sonic Hedgehog pathway in mammals.Elife 5.
pRL-TK-IRES-ZSGREEN plasmid information:
pRL-TK, pLVX-IRES-ZSGREEN1 as a commercial plasmid, was purchased from Addgene and IRES-ZSGREEN was cloned into pRL-TK to obtain pRL-TK-IRES-ZSGREEN plasmid.
2. NIH-3T3 cell sources with endogenous SMO knockdown
The plasmid pX330 and pLVX-IRES-Puro containing Smo gRNA were transfected on the basis of NIH-3T3 cells, and the endogenous Smo gene knockout monoclonal cell line was selected after puromycin selection. The cells do not express Smoothened protein and thus can be used in related studies.
Wherein the pX330 plasmid pLVX-IRES-Puro plasmid is a commercial plasmid, available commercially from Addgene.
3. Starvation medium (containing Shh-N conditional medium)
HEK293T cells were derived from ATCC, and after transfection of pHus-SHH-1-198 with the plasmid, the medium was changed to serum-free DMEM medium and culture was continued for 48 hours when the cells were substantially full. Collecting supernatant, filtering, packaging, and freezing to-80 deg.C for use. Reference may be made in particular to the following documents:
Nedelcu,D.,Liu,J.,Xu,Y.,Jao,C.,and Salic,A.(2013).Oxysterol binding to the extracellular domain of Smoothened in Hedgehog signaling.Nat Chem Biol 9,557-564.
pSMO-D473H, pSMO-E518K mutant plasmids, pGLIBS and pEGFP-N1 plasmids
First, SMO cDNA was constructed on pcDNA3.0 plasmid, and the vector was digested with EcoR1 and BamH1, and constructed by the method of enzymatic ligation. SMO-D473H and SMO-E518K were further constructed by point mutation on the basis of SMO-WT.
PGlIBS plasmid information is described in the document 1 above.
PEGFP-N1 was purchased from addgene.
RNA extraction and qPCR
Day 0, cells of NIH-3T3 were seeded in a 60-mm dish at a cell size of 1X 10 6, and cultured in 10% FCS medium for 24 hours;
day 2, the original culture medium is sucked off and replaced by DMEM medium to starve for 6 hours. Starvation medium (containing Shh-N conditional medium in DMEM) was then changed to contain different concentrations of compounds. The Hh signaling pathway is maximally activated upon stimulation by Shh-N.
Day 3 after 30 hours of incubation, cells were rinsed once with PBS, 1ml trizol Reagent (Invitrogen, 15596026) was added per plate and left to stand at room temperature for 5min; transferring the cell lysate to a precooled 1.5ml EP tube, adding 200 μl of chloroform, mixing for 20 times upside down, and standing in a 4-degree refrigerator for 10min; centrifugation at 13200rpm/4℃for 10min, transferring 400. Mu.l of supernatant into a fresh 1.5ml EP tube; adding 600 μl of isopropanol, mixing, standing in a refrigerator at 4deg.C for 10min; centrifuging at 13200 rpm/4deg.C for 10min, sucking off supernatant, adding 1ml75% ethanol, and washing the precipitate; centrifuging at 13200 rpm/4deg.C for 10min, sucking supernatant, standing at room temperature, and air drying; adding 22ul DEPC water, sucking and blowing up and down for several times to fully dissolve the precipitate, and quantifying the concentration of RNA to 1 mug/. Mu.l; by M-MLV REVERSE TRANSCRIPTASE (Promega) orIIQ RT SuperMix for qPCR (Vazyme R222-01) reverse transcribing the RNA into cDNA; using MonAmp TM FastGREEN QPCR Mix (monad MQ 10101S) kit QpcR (Quantitative Real-time PCR) was performed.
7. Others
Lipo2000 was purchased from Thermo under accession number 11668019;
mouse liver microsomes (BPI, lot number: MIC 255038);
rat liver microsomes (BPI, lot number: MIC 254035);
Human liver microsomes (XENOTECH, lot number: 1910096);
dog microsomes (XENOTECH, lot number: 1410114);
cynomolgus monkey liver microsomes (XENOTECH, lot number: 2010074).
1. Synthetic routes for intermediates I and III
In the first step, t-BuOK (potassium tert-butoxide) (25.00 g,222.8mmol,7.4 eq) was dispersed in t-BuOH (tert-butanol) (250 mL), and starting materials (3 a 'S,9a' R,9b 'S,11a' S) -9a ',11a' -dimethyl-2 ',3',3a ',3b',4',5',7',8',9',9a',9b ',10',11',11a' -tetradecanospiro [1, 3-dioxolan-2, 1 '-cyclopenta [2,1-a ] phenanthren ] -7' -one (I-1) (10.00 g,30.3mmol,1.0 eq) and CH 3 I (methyl iodide) (7.28 mL,116.9mmol,8.0 eq) were added under ice-bath, and the reaction solution was stirred at 50℃for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction solution was concentrated, poured into 200mL of ice water, extracted with ethyl acetate (ethyl acetate) (250 mL. Times.2), and the organic layers were combined and washed with water (250 mL. Times.2). The organic layer was dried over Na 2SO4, filtered and concentrated and the crude product was purified with petroleum ether: ethyl acetate=10:1 column chromatography, and sample collection gave (3 a's,3b ' r,9a ' r,9b's,11 a's) -6',6',9a ',11a ' -tetramethyl-2 ',3',3a ',3b ',4',6',7',8',9',9a ',9b ',10',11',11a ' -tetradecanospiro [1, 3-dioxolan-2, 1' -cyclopenta [1,2-a ] phenanthren ] -7' -one (I-2) (6.90 g, purity 85%, yield 54.1％).1H NMR(400MHz,CDCl3)δ5.65–5.45(m,1H),3.99–3.80(m,4H),2.53(ddd,J＝19.1,14.4,8.4Hz,2H),2.19–1.94(m,3H),1.84–1.59(m,6H),1.57–1.35(m,4H),1.31-1.23(m,7H),1.09(dd,J＝15.2,7.7Hz,1H),0.88(dd,J＝11.6,5.2Hz,6H).
In the second step LiAlH 4 (lithium aluminum hydride) (2.54 g,66.9mmol,1.5 eq) was dispersed in THF (tetrahydrofuran) (200 mL), a solution of the starting materials (3 a ' S,3b ' R,9a ' R,9b ' S,11a ' S) -6',6',9a ',11a ' -tetramethyl-2 ',3',3a ',3b ',4',6',7',8',9',9a ',9b ',10',11',11a ' -tetradecanospiro [1, 3-dioxolan-2, 1' -cyclopenta [1,2-a ] phenanthren ] -7' -one (I-2) (10 g,30.260mmol,1.0 eq) in THF (tetrahydrofuran) (100 mL) was added under ice-bath, and the reaction solution was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give white solid (3 a's,3b' r,7's,9a' r,9b's,11 a's) -6',6',9a ',11a' -tetramethyl-2 ',3',3a ',3b',4',6',7',8',9',9a',9b ',10',11',11a' -tetradecanospiro [1, 3-dioxolan-2, 1 '-cyclopenta [1,2-a ] phenanthren ] -7' -ol (I-3) (10.70 g, purity 90%, yield 59.9%), the crude product was directly fed to the next reaction .1H NMR(400MHz,CDCl3)δ5.56(dd,J＝4.6,2.9Hz,1H),3.96–3.83(m,4H),3.24(dd,J＝10.7,4.9Hz,1H),2.18–2.08(m,1H),2.01(ddd,J＝14.2,11.4,3.0Hz,1H),1.83–1.63(m,7H),1.57–1.49(m,3H),1.44–1.32(m,3H),1.26(ddd,J＝17.7,11.7,5.8Hz,1H),1.15(d,J＝6.1Hz,3H),1.07(d,J＝6.7Hz,6H),1.02–0.91(m,1H),0.86(d,J＝6.1Hz,3H).13C NMR(101MHz,CDCl3)δ149.73,119.94,119.51,77.44,77.34,77.23,77.03,76.71,65.16,64.55,51.09,50.76,45.67,41.62,36.81,36.68,34.28,31.97,31.19,30.59,27.43,27.24,23.65,22.71,21.32,20.00,14.21.
In a third step, (3 a 'S,3b' R,7'S,9a' R,9b 'S,11a' S) -6',6',9a ',11a' -tetramethyl-2 ',3',3a ',3b',4',6',7',8',9',9a',9b ',10',11',11a' -tetradecanospiro [1, 3-dioxolan-2, 1 '-cyclopenta [1,2-a ] phenanthrene ] -7' -ol (I-3) (2.00 g,5.5mmol,1.0 eq) was dissolved in a mixed solution (25 mL) of THF (tetrahydrofuran) and methanol (methanol), and Pd (OH) 2 (palladium hydroxide) (200 mg) was added under nitrogen. The reaction solution was replaced with hydrogen 3 times. The reaction mixture was stirred at room temperature for 16hrs under hydrogen atmosphere. 1 H NMR showed the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give white solid (3 a's,3b ' r,5a ' r,7's,9a ' r,9b's,11 a's) -6',6',9a ',11a ' -tetramethyl-2 ',3',3a ',3b ',4',5',5a ',6',7',8',9',9a ',9b ',10',11',11a ' -hexadecanospiro [1, 3-dioxolan-2, 1' -cyclopenta [1,2-a ] phenanthren ] -7' -ol (I-4) (1.60 g, purity 85%, yield 67.5%), the crude product was directly taken to the next step .1H NMR(400MHz,CDCl3)δ3.97–3.78(m,3H),3.21(dd,J＝11.4,4.4Hz,1H),2.09-1.92(m,1H),1.83-1.74(m,2H),1.72–1.57(m,5H),1.55–1.46(m,4H),1.40–1.17(m,6H),1.08–1.00(m,1H),1.00–0.92(m,3H),0.98-0.96(m,3H),0.85–0.81(m,4H),0.82-0.79(m,3H).
In a fourth step, (3 a ' S,3b ' R,5a ' R,7' S,9a ' R,9b ' S,11a ' S) -6',6',9a ',11a ' -tetramethyl-2 ',3',3a ',3b ',4',5',5a ',6',7',8',9',9a ',9b ',10',11',11a ' -hexadecanospiro [1, 3-dioxolan-2, 1' -cyclopenta [1,2-a ] phenanthren ] -7' -ol (I-4) (3.30 g,9.1mmol,1.0 eq) was dissolved in THF (50 mL), HCl (hydrochloric acid) was added under ice-bath (30 mL,3 mol/L). The reaction was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2). The organic layers were combined and washed with saturated sodium bicarbonate solution (30 ml×2). The organic layer was dried over anhydrous sodium sulfate and concentrated to give (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -7-hydroxy-6, 9a,11 a-tetramethylhexadeca-1H-cyclopenta [1,2-a ] phenanthren-1-one (I-5) (2.40 g, purity 70%, yield) as a white solid 57.9％).1H NMR(400MHz,CDCl3)δ3.25–3.17(m,1H),2.44(dd,J＝19.3,9.1Hz,1H),2.13–2.01(m,1H),1.97–1.86(m,2H),1.81–1.76(m,1H),1.75–1.61(m,4H),1.60–1.41(m,5H),1.40–1.16(m,5H),0.99–0.95(m,3H),0.89-0.88(m,4H),0.85(s,3H),0.81(s,3H),0.72–0.62(m,1H).
In a fifth step, (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -7-hydroxy-6, 9a,11 a-tetramethylhexadeca-hydro-1H-cyclopenta [1,2-a ] phenanthren-1-one (I-5) (2.40 g,7.5mmol,1.0 eq) was dissolved in DMF (N, N-dimethylformamide) (50 mL), imidazole (2.05 g,30.1mmol,4 eq) and TBS-Cl (t-butyldimethylchlorosilane) (3.41 g,22.6mmol,2.5 eq) were added at room temperature. The reaction was stirred at room temperature for 16hrs. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (50 mL. Times.2). The organic layers were combined and washed with water (80 ml×4). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1 column chromatography and samples were collected to give (3 as,3br,5ar,7s,9ar,9bs,11 as) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -6, 9a,11 a-tetramethylhexadeca-1H-cyclopenta [1,2-a ] phenanthren-1-one (I-6) (2.90 g, 80% purity, yield) as a white solid 71.1％).1H NMR(400MHz,CDCl3)δ3.17(dd,J＝11.0,4.6Hz,1H),2.43(dd,J＝19.2,8.5Hz,1H),2.10-2.01(m,1H),1.97–1.83(m,2H),1.81–1.75(m,1H),1.57(ddd,J＝32.2,24.0,12.5Hz,8H),1.36(dd,J＝12.9,3.6Hz,1H),1.29(dd,J＝12.5,3.8Hz,1H),1.25–1.15(m,2H),1.04(s,1H),0.98–0.91(m,1H),0.90–0.87(m,15H),0.85–0.81(m,4H),0.77(s,3H),0.69–0.60(m,1H),0.03(d,J＝2.6Hz,6H).13C NMR(101MHz,CDCl3)δ221.43,79.41,77.33,77.01,76.70,56.57,54.86,51.62,47.69,39.54,37.31,36.56,35.89,34.66,31.84,31.52,28.58,27.98,25.92,21.79,21.27,19.95,18.12,16.01,14.42,13.76,-3.78,-4.92.
In a sixth step, (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -6, 9a,11 a-tetramethylhexadeca-hydro-1H-cyclopenta [1,2-a ] phenanthren-1-one (I-6) (250 mg,0.6mmol,1.0 eq), m-CPBA (m-chloroperoxybenzoic acid) (199.38 mg,1.2mmol,2.0 eq) (25 mL), naHCO 3 (sodium bicarbonate) (73 mg,0.87mmol,1.5 eq) were dispersed in methylene chloride (methylene chloride). The reaction was stirred at room temperature for 16hrs. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. The reaction solution was washed with saturated sodium sulfite (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate and the crude product was concentrated using petroleum ether: ethyl acetate=10:1 column chromatography and samples were collected to give white solid (4 as,4br,6ar,8s,10ar,10bs,12 as) -8- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -7, 10a,12 a-tetramethyl-3, 4a,4b,5, 6a,7,8,9,10 a,10b,11,12 a-hexadeca-2H-naphtho [2,1-f ] chromen-2-one (I-7) (160 mg, purity 90% yield 55.6％).1H NMR(400MHz,CDCl3)δ7.23(s,2H),3.15(dd,J＝10.9,4.8Hz,1H),2.63(ddd,J＝19.0,8.8,2.4Hz,1H),2.57-2.45(m,1H),1.98-1.87(m,3H),1.67–1.37(m,8H),1.35-1.22(m,5H),1.20–0.93(m,3H),0.88–0.82(m,12H),0.82–0.75(m,5H),0.72(s,3H),0.03–-0.01(d,6H).13C NMR(101MHz,CDCl3)δ171.65,83.38,79.30,77.34,77.22,77.02,76.70,55.26,54.18,46.44,39.41,39.22,37.52,37.12,36.41,31.60,28.69,28.56,27.89,25.91,21.46,21.32,20.09,19.81,18.12,16.00,14.24,-3.79,-4.92.
In the seventh step LiAlH 4 (lithium aluminum hydride) (0.88 g,23.2mmol,1.5 eq) was dispersed in THF (tetrahydrofuran) (100 mL), a solution of the starting material (4 aS,4bR,6aR,8S,10aR,10bS,12 aS) -8- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -7, 10a,12 a-tetramethyl-3, 4a,4b,5, 6a,7,8,9,10 a,10b,11,12 a-hexadeca-2H-naphtho [2,1-f ] chromen-2-one (I-7) (5.2 g,11.587mmol,1.0 eq) in THF (50 mL) was added under ice-bath, and the reaction solution was stirred at room temperature for 3hrs. TLC (petroleum ether: ethyl acetate=1:1) showed the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give white solid { [ (2 s,4ar,4bs,7s,8ar,8s,10 ar) -7-hydroxy-8- (3-hydroxypropyl) -1, 4a, 7-tetramethyl decatetrahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (I-8) (3.50 g, purity 90%, yield 61.9%), the crude product was directly fed to the next reaction.
1H NMR(400MHz,dmso-d6)δ4.27(s,1H),4.02(s,1H),3.38–3.32(m,2H),3.20(dd,J＝10.4,5.1Hz,1H),1.97(d,J＝11.7Hz,1H),1.61–1.44(m,8H),1.42–1.35(m,1H),1.20(ddd,J＝41.7,29.7,19.8Hz,3H),1.08–0.96(m,4H),0.89(s,3H),0.86(d,J＝2.5Hz,11H),0.84–0.78(m,3H),0.76(s,3H),0.70(s,3H),0.63(dd,J＝16.9,6.0Hz,1H),0.03(d,J＝1.7Hz,6H).13C NMR(101MHz,CDCl3)δ79.45,77.33,77.22,77.01,76.70,74.05,61.70,55.55,54.40,51.37,42.32,40.43,39.39,37.28,36.48,33.01,32.78,28.59,28.00,25.91,23.94,22.22,21.73,21.43,18.12,16.03,14.34.
In the eighth step { [ (2S, 4aR,4bS,7S,8aR,8S,10 aR) -7-hydroxy-8- (3-hydroxypropyl) -1, 4a, 7-tetramethyl decatetrahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (I-8) (3.50 g,7.9mmol,1.0 eq) was dissolved in dichloromethane (150 mL), and the reaction mixture was stirred at room temperature for 3hrs while adding DMAP (0.19 g,1.5mmol,0.2 eq), triethylamine (5.37 mL,38.6mmol,5.0 eq) and Ac2O (1.2 mL,12.8mmol,1.7 eq) under an ice bath. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was washed with water (100 mL. Times.2) and saturated sodium bicarbonate (100 mL). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=5:1 column chromatography and samples were collected to give acetic acid-3- [ (1 s,2s,4as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2-hydroxy-2, 4b, 8-tetramethyl deca-hydrophenanthreyl ] propyl ester (I-9) as a white solid (3.20 g, purity 90%, yield 75.3％).1H NMR(400MHz,CDCl3)δ4.10–3.93(m,2H),3.19–3.08(m,1H),2.01(s,3H),1.98–1.90(m,1H),1.76–1.40(m,10H),1.15-1.32(m,3H)1.02(s,3H),0.85(s,12H),0.80–0.73(m,4H),0.72(s,3H),0.00(d,J＝1.5Hz,6H).13C NMR(101MHz,CDCl3)δ171.25,79.44,77.33,77.21,77.01,76.69,73.40,64.88,55.58,54.31,53.74,42.58,40.03,39.39,37.30,36.52,33.16,30.11,29.70,28.59,28.00,25.91,24.63,22.25,21.73,21.15,21.05,18.12,16.03,14.34,-3.79,-4.92.
Triphenylphosphine (2.54 g,9.7mmol,1.5 eq) was dissolved in dichloromethane (100 mL), iodine (2.46 g,9.7mmol,1.5 eq) was added at room temperature, and acetic acid-3- [ (1S, 2S,4aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2-hydroxy-2, 4b, 8-tetramethyl deca-hydrophenanthreyl ] propyl ester (I-9) (3.20 g,6.5mmol,1.0 eq) was added after stirring the reaction solution at room temperature for 5 min. The reaction was stirred at room temperature for 16h, tlc (petroleum ether: ethyl acetate=10:1) indicated that the reaction was complete. The reaction was quenched with saturated sodium sulfite and washed with saturated sodium bicarbonate (100 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1 column chromatography, and samples were collected to give acetic acid-3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (I-10) as a white solid (1.10 g, purity 90%, yield 42.2％).1H NMR(400MHz,CDCl3)δ4.10–3.97(m,2H),3.23(dd,J＝11.5,4.6Hz,1H),2.20-2.10(m,2H),2.06(s,3H),2.02–1.84(m,3H),1.82-1.65(m,6H),1.60(s,3H),1.49(s,2H),1.43(s,1H),1.38(dd,J＝13.0,3.6Hz,1H),1.33–1.27(m,1H),1.12-1.07(m,2H),1.02–0.94(m,4H),0.92–0.83(m,4H),0.82(s,3H),0.79(s,3H).
In a tenth step acetic acid-3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (I-10) (600 mg,1.6mmol,1.0 eq) was dissolved in DMF (N, N-dimethylformamide) (50 mL), and imidazole (450 mg,6.6mmol,4.0 eq) and TBS-Cl (t-butyldimethylsilyl chloride) (748 mg,5.0mmol,5.0 eq) were added at room temperature. The reaction was stirred at room temperature for 16hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete and the reaction was extracted with ethyl acetate (50 ml×2). The organic layers were combined and washed with water (80 ml×4). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1 was passed through the column and the sample was collected and taken directly into the next step.
The product of the previous step was dissolved in methanol (methanol) (20 mL), an aqueous solution (3 mL) of 20% NaOH (sodium hydroxide) was added under ice bath, and the reaction solution was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (50 mL), and washed with water (40 mL. Times.3). The organic layer was concentrated to give 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propan-1-ol (I-11) as a white solid (400 mg, purity 90%, two-step yield 55.6％).1H NMR(400MHz,CDCl3)δ3.57–3.50(m,2H),3.08(dd,J＝11.2,4.7Hz,1H),1.95(m,6H),1.53-1.47(m,9H),1.37(s,5H),1.27-1.20(m,1H),0.99–0.87(m,2H),0.82-0.80(m 18H),0.79(s,8H),0.78(s,3H),0.71(s,3H),0.65(s,3H).
In the eleventh step, DMP (dess-Martin reagent) (2.54 g,6.0mmol,2.0 eq) was dispersed in methylene chloride (methylene chloride) (5 mL), and a methylene chloride solution (10 mL) of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propan-1-ol (I-11) (1.30 g,3.0mmol,1.0 eq) was added under ice-bath, and the reaction solution was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction mixture was diluted with dichloromethane (50 mL) and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=15:1, and concentrated to give 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (1.10 g, 90% purity, yield) as a colorless solid 76.5％).1H NMR(400MHz,CDCl3)δ9.73(s,1H),3.15(dd,J＝11.2,4.7Hz,1H),2.51–2.25(m,2H),2.12-2.00(m,2H),1.95-1.83(m,2H),1.71–1.47(m,9H),1.46–1.21(m,4H),1.01-0.95(m,3H),0.85(s,12H),0.80–0.75(m,4H),0.72(s,3H),0.00(d,J＝2.8Hz,6H).
In the twelfth step, 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (1.10 g,2.5mmol,1.0 eq) was dissolved in t-BuOH (t-butanol) (20 mL), an aqueous solution (5 mL) of 2-methyl-2-butene (1.07 g,15.2mmol,6.0 eq) and sodium dihydrogen phosphate (0.91 g,7.6mmol,3.0 eq) were added under ice bath, and the reaction mixture was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (20 mL), and washed with saturated sodium bicarbonate (20 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=3:1, and concentrated to give 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) as a white solid (1.10 g, purity 90%, yield 86.7％).1H NMR(400MHz,MeOD)δ3.19(dd,J＝11.1,4.7Hz,1H),2.10-2.40(m,4H),2.00–1.82(m,3H),1.72–1.56(m,6H),1.56–1.35(m,5H),1.30–1.21(m,4H),1.07–0.92(m,3H),0.80-0.85(m,10H),0.79(s,3H),0.73(s,3H),0.00(d,J＝3.1Hz,6H).13C NMR(101MHz,MeOD)δ176.06,131.62,127.93,79.57,54.38,54.20,48.23,48.02,47.80,47.59,47.38,47.17,46.95,39.09,37.79,36.63,36.12,33.21,32.91,32.00,27.88,27.77,25.03,24.28,21.83,21.55,20.13,18.21,17.58,15.11,13.44,-4.95.
In the thirteenth step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (0.50 g,1.1mmol,1.0 eq) was dissolved in tetrahydrofuran (10 mL), and a tetrahydrofuran solution of TBAF (10.0 mL,10.0mmol,9.0 eq) was added thereto, and the reaction mixture was stirred overnight at 60 ℃. TLC (petroleum ether: ethyl acetate=2:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (30 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and the crude product was purified by passing methanol=10:1 through a column to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionic acid (III) (187.3 mg, yield=50.98%, purity ＝96.7％).1H NMR(400MHz,DMSO)δ12.00(s,1H),4.32(d,J＝4.8Hz,1H),3.33(s,1H),3.00(dd,J＝12.2,8.0Hz,1H),2.50(s,2H),2.37–2.15(m,3H),2.15–2.02(m,2H),2.02–1.77(m,3H),1.67–1.46(m,7H),1.02–0.87(m,5H),0.83–0.73(m,5H),0.72–0.64(m,3H).
2. Intermediate II & IV synthetic route
The first step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propan-1-ol (I-11) (6 g,13.800mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (100 mL), and after adding methanesulfonyl chloride (4.74 g, 24.84mmol, 1.8 eq), 4-Dimethylaminopyridine (DMAP) (1.35 g,11.040mmol,0.8 eq) and N, N-Diisopropylethylamine (DIEA) (4.561 mL,27.600mmol,2.0 eq), the mixture was stirred at 40℃for 4hr; TLC (petroleum ether: ethyl acetate=2:1) monitoring the reaction showed no starting material remaining, with product formation; dichloromethane (30 mL) was added to the reaction solution, which was washed with saturated aqueous sodium hydrogencarbonate (100 mL) and water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 19:1) to give 4-methylbenzenesulfonic acid-3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl ester (II-1) (5 g,7.640mmol, 55.37%) as a white solid .1H NMR(400MHz,CDCl3)δ7.80(d,J＝8.3Hz,2H),7.35(d,J＝8.0Hz,2H),4.01(dd,J＝6.4,3.3Hz,2H),3.17(dd,J＝11.1,4.7Hz,1H),2.45(s,3H),2.13–1.98(m,4H),1.85(d,J＝11.3Hz,3H),1.77–1.66(m,3H),1.65–1.57(m,4H),1.53(s,3H),1.37–1.29(m,2H),1.03–0.93(m,3H),0.89(s,12H),0.77(s,3H),0.74(s,3H),0.03(d,J＝2.7Hz,6H).
The second step compound 4-methylbenzenesulfonic acid-3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (II-1) (5 g, 8.4819 mmol,1.0 eq) was added to a reaction flask containing dimethyl sulfoxide (DMSO) (50 mL), sodium cyanide (1.66 g,33.958mmol,4.0 eq) was added, and the mixture was warmed to 40℃and stirred for 16hr. TLC (petroleum ether: ethyl acetate=20:1) monitoring the reaction showed complete reaction of the starting materials; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (20 ml×3), the organic phases were combined, washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 97:3) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanenitrile (II-2) (3.5 g,7.098mmol, 83.61%) as a white solid .1H NMR(400MHz,CDCl3)δ3.18(dd,J＝11.1,4.7Hz,1H),2.39–2.15(m,5H),2.13–2.04(m,2H),1.91(d,J＝10.8Hz,3H),1.80–1.69(m,3H),1.64(s,2H),1.62(s,3H),1.59(s,1H),1.34(d,J＝3.2Hz,1H),1.01(dd,J＝17.9,4.5Hz,2H),0.89(s,10H),0.84(d,J＝6.6Hz,3H),0.81(s,3H),0.76(d,J＝7.6Hz,3H),0.04(d,J＝2.8Hz,6H)
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanenitrile (II-2) (3 g,6.760mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (40 mL), cooled to-50-40℃under nitrogen protection, and then diisobutylaluminum hydride (8.112 mL,8.112mmol,1.2 eq) (1.0M solution in hexanes) was added and stirred for 1hr; TLC (petroleum ether: ethyl acetate=20:1) monitored disappearance of starting material; the reaction solution was quenched by dropwise addition of saturated aqueous ammonium chloride (10 mL) at-40 ℃ and extracted with dichloromethane (20 mL x 3), the organic phases were combined, washed with water (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 49:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanal (II-3) (1.4 g,2.820mmol, 41.72%) as a colorless oil .1H NMR(400MHz,CDCl3)δ9.77(s,1H),3.18(dd,J＝11.2,4.6Hz,1H),2.40(t,J＝7.2Hz,2H),2.19–2.05(m,4H),2.01–1.88(m,3H),1.78–1.65(m,4H),1.59(s,3H),1.54(t,J＝9.7Hz,3H),1.48–1.28(m,2H),1.05(d,J＝16.5Hz,2H),0.88(d,J＝4.7Hz,13H),0.81(s,3H),0.76(d,J＝7.9Hz,3H),0.03(d,J＝2.8Hz,6H).
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanal (II-3) (1.4 g,3.133mmol,1.0 eq) was added to a reaction flask containing t-butanol (20 mL), and after adding 2-methyl-2-butene (1.32 g,18.80 mmol,6.0 eq) at 0℃water (5 mL) of sodium dihydrogen phosphate (1.13 g, 9.mmol, 3.0 eq) and sodium chlorite (0.99 g,10.967mmol,3.5 eq) was further added and stirred for 2hr at room temperature; TLC (petroleum ether: ethyl acetate=1:1) (T1) (dichloromethane: methanol=20:1) (T2) showed disappearance of starting material and formation of a dot with increased polarity; after the reaction solution was concentrated under reduced pressure (water pump, 50 ℃) and most of the tert-butanol was removed, water (20 mL) was added, extracted with ethyl acetate (20 mL), the organic phases were combined, spin-dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (dichloromethane: methanol=1:0 to 24:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanoic acid (II) (900 mg,1.750mmol, 55.86%) as a white solid .1H NMR(400MHz,DMSO)δ11.95(s,1H),3.19(m,1H),2.12(m,3H),2.04–1.79(m,6H),1.60(m,6H),1.52(s,3H),1.48(m,2H),1.36–1.24(m,2H),0.97(m,2H),0.84(s,12H),0.74(s,3H),0.69(s,3H),0.00(d,J＝3.3Hz,6H).
In a fifth step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (200 mg,0.432mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), tetrabutylammonium fluoride (1M in THF) (2 mL) was added at room temperature, warmed to 50℃and stirred overnight, TLC and LC-MS were examined for reaction, after completion of the reaction water was added thereto to kill, then extracted (50 mL) with ethyl acetate, the organic phase was washed twice with saturated sodium chloride, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 0:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (77 mg,0.221mmol, yield: 51.16) as a white solid compound.
LC-MS(ESI)[M-H]-＝347.2.
3. Intermediate V synthetic route
The first step compound 2-methylpyridine (3 g,32.2mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (30 mL), cooled to-78 ℃ under nitrogen protection, then lithium diisopropylamide (19.3 mL,38.6mmol,1.2eq,2 m) was added dropwise, stirred for 0.5hr, a solution of 6-bromohexan-1-ol (7.58 g,41.8mmol,1.3 eq) in anhydrous tetrahydrofuran (10 mL) was added, stirring was resumed to room temperature (20 ℃) for 16hr, tlc (petroleum ether: ethyl acetate=2:1) was monitored to generate a new spot of increased reaction polarity (R f =0.25), (10 mL) was quenched, then water (10 mL) was added, extracted with ethyl acetate (15 mL 3), the organic phases were combined, washed with water (20 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying (water pump, 40 ℃) to give a yellow liquid. The crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:4) and the product at R f =0.25 was collected to give the product 7- (pyridin-2-yl) hept-1-ol (V-1) (1.60 g,8.27mmol, 25.7%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.53(d,J＝4.2Hz,1H),7.62(td,J＝7.7,1.5Hz,1H),7.20–7.09(m,2H),3.63(t,J＝6.6Hz,2H),2.85–2.77(m,2H),1.73(dd,J＝14.8,7.2Hz,2H),1.57(dd,J＝13.3,6.6Hz,2H),1.37(d,J＝3.3Hz,6H).
In a second step, 7- (pyridin-2-yl) heptan-1-ol (V-1) (800 mg,4.13mmol,1.0 eq) was added to a reaction flask containing acetonitrile (20 mL), triphenylphosphine (2.17 g,8.27mmol,2.0 eq) and carbon tetrabromide (2.74 g,8.27mmol,2.0 eq) were added at 0deg.C, and after returning to room temperature (18 ℃) and stirring was continued for 16hr, TLC (petroleum ether: ethyl acetate=2:1) was monitored to show disappearance of starting material. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil, which was extracted with water (15 mL), dichloromethane (10 ml×3), the organic phases were combined, washed with water (20 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃). The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1), separated and the new spot (R f =0.50) was collected to give the product 2- (7-bromoheptyl) pyridine (V-2) (720 mg,2.81mmol, 67.9%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.58–8.50(m,1H),7.65(td,J＝7.7,1.5Hz,1H),7.22–7.11(m,2H),3.40(t,J＝6.8Hz,2H),2.88–2.78(m,2H),1.90–1.80(m,2H),1.74(dd,J＝14.9,7.2Hz,2H),1.41(ddd,J＝13.7,10.0,5.6Hz,6H).
The third step compound 2- (7-bromoheptyl) pyridine (V-2) (720 mg,2.81mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (7 mL), and after adding potassium phthalimide (6278 mg,3.37mmol,1.2 eq), stirred at room temperature (18 ℃) for 16hr, and TLC (Petroleum ether: ethyl acetate=2:1) monitored to show the completion of the reaction of the starting material. The reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil. Water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1), and the new spot (R f =0.20) was collected to give the product 2- [7- (pyridin-2-yl) heptyl ] isoindole-1, 3-dione (V-3) (800 mg,1.94mmol, 69.2%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.58–8.50(m,1H),7.65(td,J＝7.7,1.5Hz,1H),7.22–7.11(m,2H),3.40(t,J＝6.8Hz,2H),2.88–2.78(m,2H),1.90–1.80(m,2H),1.74(dd,J＝14.9,7.2Hz,2H),1.41(ddd,J＝13.7,10.0,5.6Hz,6H).
The fourth step compound 2- [7- (pyridin-2-yl) heptyl ] isoindole-1, 3-dione (V-3) (200 mg,0.620mmol,1.0 eq) was added to a reaction flask containing ethanol (3 mL), hydrazine hydrate (0.060 mL,1.24mmol,2.0 eq) was added and then heated to 80 ℃ and stirred for 2hr, lc-MS monitored for basic reaction completion, cooled and filtered, the filtrate was dried under reduced pressure (water pump, 50 ℃) to give a pale yellow oil, water (10 mL) was added, and the mixture was extracted with a mixed solution of chloroform and isopropanol (chloroform: isopropanol=3:1, 8mL 5), the organic phase was dried under reduced pressure, and the oil pump was pulled dry to give 7- (pyridin-2-yl) hept-1-amine (V) (140 mg, 0.234 mmol, 58.6%) as a pale yellow oil .1H NMR(400MHz,MeOD)δ8.43(d,J＝4.7Hz,1H),8.22(dd,J＝5.9,3.3Hz,1H),7.81(dd,J＝6.0,3.3Hz,1H),7.76(dd,J＝7.7,1.7Hz,1H),7.32(d,J＝7.8Hz,1H),7.26(dd,J＝7.1,5.4Hz,1H),2.88–2.74(m,4H),1.79–1.66(m,2H),1.65–1.55(m,2H),1.38(d,J＝9.3Hz,6H).
Example 1
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3- { methyl [3- (pyridin-2-yl) propyl ] amino } propyl) propanamide
In a first step, the reaction 3- (pyridin-2-yl) propan-1-ol (1 g,7.290 mmol), dess-martin oxidant (30.92 g,72.897 mmol) and solvent dichloromethane (15 mL) were added to a 100mL round bottom flask and the reaction stirred at room temperature for 2 hours. LC-MS monitoring. After the reaction was completed, it was diluted with 80mL of methylene chloride, quenched by 1:1 addition of saturated sodium thiosulfate solution (100 mL) and saturated aqueous sodium bicarbonate solution (100 mL), extracted with methylene chloride (3X 100 mL) and concentrated in vacuo to give the crude product. The crude product was purified on silica gel (dichloromethane/methanol=15/1) to give 3- (pyridin-2-yl) propanal (1-1) (260 mg, 1.6271 mmol, 22.23%) as a black solid. LC-MS (ESI) [ m+h ] + =136.1
In a second step, reactant 3- (pyridin-2-yl) propanal (1-1) (100 mg,0.740 mmol), { [3- (methylamino) propyl ] amino } methanoic acid-2-methylpropan-2-yl ester (167.14 mg,0.88 mmol), acetic acid (0.148 mL,0.148 mmol), and solvent dichloromethane (3 mL) were added to a 50mL round bottom flask, the reaction was stirred at room temperature under nitrogen for 2 hours, and then sodium borohydride acetate (312.10 mg,1.480 mmol) was added and the reaction was stirred at room temperature overnight. LC-MS monitoring. To the reaction was added water (10 mL), extracted with dichloromethane (3×15 mL), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated in vacuo to give the crude product, which was purified by column (dichloromethane/methanol=5/1) over silica gel to give [ (3- { methyl [3- (pyridin-2-yl) propyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (1-2) (130 mg,0.412mmol, 55.65%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =308.1
In a third step, the reaction [ (3- { methyl [3- (pyridin-2-yl) propyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (1-2) (125 mg,0.407 mmol), dioxane hydrochloride solution (2 mL) and solvent methanol (2 mL) were added to a 50mL round bottom flask and stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction was completed, the reaction mixture was concentrated in vacuo to give 3- { methyl [3- (pyridin-2-yl) propyl ] amino } propan-1-amine hydrochloride (1-3) (100 mg,0.406mmol, 99.88%) as a brown oily substance, and the crude product was directly poured downward. LC-MS (ESI) [ m+h ] + =208.2
In a fourth step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylhydrophenanthrene ] butanoic acid (I) (60 mg,0.134 mmol), 3- { methyl [3- (pyridin-2-yl) propyl ] amino } propan-1-amine hydrochloride (1-3) (32.25 mg,0.156 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (73.95 mg,0.194 mmol), N, N-diisopropylethylamine (0.064 mL,0.389 mmol) and solvent N, N-dimethylformamide (1.5 mL) were added to a 5mL round bottom flask, and the reaction mixture was stirred at room temperature and the MS was monitored. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=12/1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [2- (1-methylindol-5-yl) ethyl ] butanamide (1-4) as a pale yellow solid (34.5 mg,0.047mmol, 34.97%). LC-MS (ESI) [ m+h ] + = 638.5
In a fifth step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] butanamide (1-4) (41 mg,0.064 mmol), tetrabutylammonium fluoride (168.01 mg,0.643 mmol) and the solvent tetrahydrofuran (2 mL) were added to a 25mL round bottom flask, and the reaction mixture was stirred at 50℃for 28 hours and monitored by LC-MS. After the reaction was completed, 15mL of ethyl acetate was added for dilution, and the mixture was washed 3 times with 20mL of water. The product was dissolved in water. The aqueous phase was dried by spin-drying, washed with dichloromethane/methanol=5/1, filtered and concentrated to give the crude product. Preparation of the crude product from formic acid to give 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3- { methyl [3- (pyridin-2-yl) propyl ] amino } propyl) propanamide as a white solid (1)(6.48mg,0.011mmol,17.67％).1H NMR(400MHz,DMSO)δ8.46(d,J＝4.1Hz,1H),8.27(s,1H),7.82(s,1H),7.68(t,J＝7.6Hz,1H),7.24(d,J＝7.7Hz,1H),7.22–7.14(m,1H),3.02(dt,J＝15.4,6.9Hz,3H),2.75–2.69(m,2H),2.46–2.43(m,2H),2.26(s,3H),2.10(dd,J＝18.8,11.3Hz,3H),2.01–1.78(m,7H),1.66–1.44(m,11H),1.30(dd,J＝30.9,18.4Hz,3H),0.98–0.91(m,2H),0.87(d,J＝5.9Hz,3H),0.76(d,J＝10.9Hz,5H),0.68(d,J＝6.5Hz,3H).13C NMR(101MHz,DMSO)δ172.33,161.59,149.40,136.88,132.69,127.18,123.19,121.67,77.37,56.61,54.64,54.35,53.95,41.60,38.91,37.90,37.03,36.53,35.48,35.22,33.51,32.14,28.64,27.88,26.59,26.38,25.55,22.03,21.86,19.74,16.27,14.56,14.39.LC-MS(ESI)[M+H]+＝524.4.
Example 2
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylpyrrolio [2,3-b ] pyridin-5-yl) butanamide
In the first step, the reactant 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108 mmol), 1-methylpyrrolo [2,3-b ] pyridin-5-amine (19.08 mg,0.130 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (61.62 mg,0.162 mmol), N, N-diisopropylethylamine (0.054 mL,0.324 mmol) was added to solvent N, N-dimethylformamide (3 mL) and the reaction mixture was stirred at room temperature and monitored by LC-MS. After completion of the reaction, water (15 mL) and ethyl acetate (3X 10 mL) were added thereto and extracted. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: petroleum ether: ethyl acetate=3/1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrazolo [2,3-b ] pyridin-5-yl) butanamide (2-1) (56 mg,0.095mmol, 87.56%) as a white solid. LC-MS (ESI) [ m+h ] + = 592.4.
In a second step, the reactant 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrrolio [2,3-b ] pyridin-5-yl) butanamide (2-1) (56 mg,0.095 mmol), tetrabutylammonium fluoride (0.946 mL,0.946 mmol) was added to the solvent tetrahydrofuran (3 mL) and the reaction mixture stirred at 50℃for 28 hours under nitrogen atmosphere, as monitored by LC-MS. After completion of the reaction, ethyl acetate (15 mL) was added thereto for dilution, and the mixture was washed with water (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=15/1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methylpyrrolo [2,3-b ] pyridin-5-yl) butanamide as a white solid (2)(23.96mg,0.046mmol,48.97％).1H NMR(400MHz,DMSO)δ9.89(s,1H),8.31(s,1H),8.25(d,J＝1.8Hz,1H),7.47(d,J＝3.2Hz,1H),6.41(d,J＝3.3Hz,1H),4.32(s,1H),3.79(s,3H),3.00(s,1H),2.30(t,J＝6.9Hz,2H),2.08(dd,J＝23.8,15.1Hz,2H),1.97–1.82(m,3H),1.69–1.47(m,10H),1.23(s,3H),1.05–0.92(m,2H),0.87(s,3H),0.76(s,5H),0.68(s,3H).13C NMR(101MHz,DMSO)δ171.63,144.71,136.33,133.02,131.06,129.48,127.02,119.80,119.75,99.07,77.38,54.36,53.98,37.99,37.04,36.53,36.50,33.53,32.22,31.33,28.63,27.90,24.56,22.08,21.90,19.83,16.27,14.57.LC-MS(ESI)[M+H]+＝478.3.
Example 3
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylbenzo [ d ] imidazol-5-yl) butanamide
In a first step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (60 mg,0.130 mmol), 1-methylbenzo [ d ] imidazol-5-amine (22.90 mg,0.156 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (73.95 mg,0.194 mmol), N, N-diisopropylethylamine (0.064 mL,0.389 mmol) and solvent N, N-dimethylformamide (1.5 mL) were added to a 5mL round bottom flask, and the reaction solution was stirred overnight at room temperature and LC-MS monitored. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=12/1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methylbenzo [ d ] imidazol-5-yl) butanamide (3-1) as a pale yellow solid (52.5 mg,0.089mmol, 68.41%). LC-MS (ESI) [ m+h ] + = 592.4.
In a second step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylbenzo [ d ] imidazol-5-yl) butanamide (7-1) (65 mg,0.110 mmol), tetrabutylammonium fluoride (287.11 mg,1.098 mmol) and solvent tetrahydrofuran (2 mL) were added to a 25mL round bottom flask, and the reaction mixture was stirred at 50℃for 28 hours and monitored by LC-MS. After the reaction was completed, 15mL of ethyl acetate was added for dilution, and the mixture was washed 3 times with 20mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. Purification of the crude product by preparative plate (dichloromethane/methanol=10/1) gives 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylbenzo [ d ] imidazol-5-yl) butyramide as a white solid (3)(31.52mg,0.063mmol,57.27％).1H NMR(400MHz,DMSO)δ9.80(s,1H),8.10(s,1H),7.99(d,J＝1.5Hz,1H),7.45(d,J＝8.7Hz,1H),7.39(d,J＝1.7Hz,1H),4.31(d,J＝5.1Hz,1H),3.80(s,3H),3.03–2.96(m,1H),2.29(s,2H),2.24–1.78(m,7H),1.78–1.37(m,12H),1.36–1.21(m,2H),1.02–0.96(m,1H),0.87(s,3H),0.77(s,3H),0.68(s,3H).13C NMR(101MHz,DMSO)δ171.26,145.38,143.81,134.35,133.06,131.36,126.99,115.90,110.23,110.18,77.39,54.37,53.99,38.93,38.00,37.05,36.73,36.54,33.54,32.24,31.13,28.64,27.90,24.60,22.09,21.90,19.83,16.27,14.57.LC-MS(ESI)[M+H]+＝478.3.
Example 4
Preparation of the Compound 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] propionamide
In a first step, 1H-indole-5-carbaldehyde (1.5 g,10.333 mmol) as a reactant and N, N-dimethylformamide (3 mL) as a solvent were charged in a 50mL three-necked flask, the reaction mixture was placed in an ice bath to be filled with N2, sodium hydrogen (0.50 g,20.667 mmol) was added at 0℃and stirred for 30 minutes, and methyl iodide (2.93 g,20.667 mmol) was added at 0℃and the reaction mixture was stirred at room temperature for 1 hour and monitored by LC-MS. After the reaction was completed, 70mL of water was added thereto, and the mixture was extracted with ethyl acetate (3X 50 mL) and concentrated in vacuo to give a crude product. The crude product was purified by column chromatography over silica gel (petroleum ether: ethyl acetate=3/1) to give 1-methylindole-5-carbaldehyde (4-1) as a yellow solid (1.2 g,7.538mmol, 72.95%). LC-MS (ESI) [ m+h ] + =160.1
In a second step, the reaction 1-methylindole-5-carbaldehyde (4-1) (1 g, 6.284 mmol), ammonium acetate (0.97 g,12.564 mmol) and solvent acetic acid (3 mL), nitromethane (15 mL) were placed in a 100mL round bottom flask, and the reaction was stirred under nitrogen at 80℃for 8 hours and monitored by LC-MS. The reaction mixture was concentrated in vacuo to give crude product, which was purified by silica gel column (petroleum ether: ethyl acetate=5/1) to give 1-methyl-5- [ (1E) -2-nitrovinyl ] indole (4-2) (630 mg,2.763mmol, 43.98%) as a yellow solid. LC-MS (ESI) [ m+h ] + =203.1
In a third step, the reaction 1-methyl-5- [ (1E) -2-nitrovinyl ] indole (4-2) (200 mg,0.989 mmol) and solvent tetrahydrofuran (5 mL) were charged to a 50mL round bottom flask, the system was purged with nitrogen, and lithium aluminum hydride (93.84 mg,2.473 mmol) was added at 0deg.C. The reaction solution was stirred under nitrogen atmosphere for 5 hours. LC-MS monitoring. The reaction mixture was quenched with sodium sulfate decahydrate, filtered, washed and concentrated in vacuo to give crude product, which was purified by silica gel column (dichloromethane/methanol=4/1) to give 2- (1-methylindol-5-yl) ethan-1-amine (4-3) (100 mg,0.492mmol, 49.75%) as pale yellow oily substance. LC-MS (ESI) [ m+h ] + =175.1
In a fourth step, the reaction 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (60 mg,0.134 mmol), 2- (1-methylindol-5-yl) ethan-1-amine (4-3) (27.96 mg,0.160 mmol), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (76.26 mg,0.201 mmol), N, N-diisopropylethylamine (51.84 mg,0.401 mmol) and solvent N, N-dimethylformamide (3 mL) were added to a 25mL round bottom flask, and the reaction solution was stirred overnight at room temperature and LC-MS monitored. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: petroleum ether: ethyl acetate=3/1) to give 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] propionamide (4-4) as a pale yellow solid (45 mg,0.071mmol, 52.85%).
LC-MS(ESI)[M+H]+＝605.4
In a fifth step, the reaction 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] propionamide (4-4) (45 mg,0.071 mmol), tetrabutylammonium fluoride (194.48 mg,0.744 mmol) and solvent tetrahydrofuran (2 mL) were charged in a 25mL round bottom flask, and the reaction stirred at 50deg.C for 28 hours, monitored by LC-MS. After the reaction was completed, 15mL of ethyl acetate was added for dilution, and the mixture was washed 3 times with 20mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. Purification of the crude product by preparative plate (dichloromethane/methanol=10/1) gives 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] propionamide as a white solid (4)(5.57mg,0.011mmol,14.96％).1H NMR(400MHz,CDCl3)δ7.43(s,1H),7.29(s,1H),7.12–7.00(m,2H),6.43(d,J＝2.8Hz,1H),5.42(s,1H),3.79(s,3H),3.62–3.50(m,2H),3.21(dd,J＝11.5,4.6Hz,1H),2.90(t,J＝6.7Hz,2H),2.36(ddd,J＝19.3,13.3,5.5Hz,2H),2.24–1.99(m,4H),1.85(d,J＝16.8Hz,2H),1.71–1.61(m,6H),1.33–1.24(m,6H),1.01–0.94(m,4H),0.91–0.83(m,2H),0.79(d,J＝7.3Hz,6H).13C NMR(101MHz,CDCl3)δ135.68,131.81,129.55,129.23,128.77,128.37,122.53,120.58,109.43,100.56,54.25,53.90,41.03,38.82,37.94,36.92,36.48,35.75,35.64,33.48,32.90,32.08,29.78,29.33,28.05,27.62,27.22,25.23,21.87,21.74,19.51,15.37,14.26.LC-MS(ESI)[M+H]+＝491.4.
Example 5
Synthesis of Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [1- (6-phenylhexyl) -1,2, 3-triazacyclopentan-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
Reference example 93 conversion of hept-6-yn-1-ol to hex-5-yn-1-ol gave the compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [1- (6-phenylhexyl) -1,2, 3-triazacyclopent-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (19 mg,0.036mmol, yield) ：92.33％).1H NMR(400MHz,CDCl3)δ7.28(d,J＝7.4Hz,1H),7.24(s,2H),7.17(dd,J＝12.3,7.2Hz,3H),4.30(t,J＝7.1Hz,2H),3.23(dd,J＝11.5,4.6Hz,1H),2.94–2.73(m,1H),2.70–2.56(m,3H),2.43(dd,J＝16.3,7.0Hz,2H),2.23(d,J＝8.9Hz,1H),2.00(s,2H),1.91–1.85(m,3H),1.79–1.75(m,1H),1.71(d,J＝15.0Hz,2H),1.66–1.61(m,3H),1.58(s,3H),1.44–1.24(m,7H),1.08–0.99(m,2H),0.98(d,J＝4.9Hz,3H),0.90(dd,J＝21.1,6.6Hz,2H),0.81(d,J＝10.6Hz,6H).13C NMR(101MHz,CDCl3)δ142.40,132.10,128.38,128.31,128.20,125.73,120.46,79.10,54.30,53.94,50.33,38.84,37.97,36.96,36.53,35.77,33.52,32.17,31.16,30.30,29.04,28.59,28.06,27.65,26.36,24.56,24.05,21.93,21.84,19.62,15.39,14.31.LC-MS(ESI)[M+1]+＝518.
Example 6
Preparation of the Compound 4- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylpyrrolio [3,2-b ] pyridin-5-yl) butanamide
In a first step, the reaction mixture, 5-bromo-1H-pyrrolo [3,2-b ] pyridine (1 g,5.075 mmol) and solvent N, N-dimethylformamide (20 mL), were placed in a 50mL three-necked flask, the reaction mixture was placed in an ice bath under nitrogen protection, sodium hydride (0.37 g,15.226 mmol) was added at 0℃and stirred for 30 minutes, methyl iodide (1.44 g,10.151 mmol) was added at 0℃and the reaction mixture was stirred at room temperature for 1.5 hours and monitored by LC-MS. After the reaction was completed, 70mL of water was added thereto, and the mixture was extracted with ethyl acetate (3X 50 mL) and concentrated in vacuo to give a crude product. The crude product was purified by column chromatography over silica gel (petroleum ether: ethyl acetate=3/1) to give 5-bromo-1-methylpyrrolo [3,2-b ] pyridine (6-1) (813 mg, 3.850 mmol, 75.90%) as a yellow solid.
In a second step, the reaction 5-bromo-1-methylpyrrolo [3,2-b ] pyridine (6-1) (180.19 mg,2.772 mmol), cuprous oxide (89.45 mg,0.370 mmol), L-proline (638.22 mg,5.543 mmol) and solvent water (3 mL) and dimethyl sulfoxide (12 mL) were added to a 50mL round bottom flask, and the reaction was stirred under nitrogen at 100deg.C for one hour. The reaction solution was diluted with ethyl acetate and water. The aqueous layer was extracted with ethyl acetate (3X 50 mL). The ethyl acetate layers were combined and washed with saturated brine (1×50 mL). The ethyl acetate layer was dried over Na2SO4, filtered and concentrated. The crude product was purified on silica gel (dichloromethane/methanol=20/1) to give 5-azido-1-methylpyrrolo [3,2-b ] pyridine (6-2) (120 mg,0.686mmol, 37.12%) as a white solid.
LC-MS(ESI)[M+H]+＝174.1.
In the third step, 5-azido-1-methylpyrrolo [3,2-b ] pyridine (6-2) (100 mg,0.577 mmol) was dissolved in methanol (15 mL), stannous chloride (1094.81 mg,5.774 mmol) was added and placed under nitrogen protection, concentrated hydrochloric acid (5 mL) was added dropwise, and the mixture was heated at 70℃under reflux overnight. After the completion of the reaction, the reaction mixture was returned to room temperature, and then the pH was adjusted to 8 with a saturated aqueous sodium hydrogencarbonate solution. The organic phase was separated three times using ethyl acetate (25 mL) and dried to give crude product which was purified by column chromatography over silica gel (dichloromethane/methanol=10/1) to give 1-methylpyrrolo [3,2-b ] pyridin-5-amine (6-3) (26 mg,0.177mmol, 30.59%) as a white product. LC-MS (ESI) [ m+h ] + =148.1.
In the fourth step 1-methylpyrrolo [3,2-b ] pyridin-5-amine (6-3) (20 mg,0.136 mmol), 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (69.18 mg,0.149 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (77.51 mg,0.204 mmol) was dissolved in N, N-dimethylformamide (2 mL), and N, N-diisopropylethylamine was added, and the reaction solution was stirred at room temperature overnight and monitored by LC-MS. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: petroleum ether: ethyl acetate=3/1) to give the product 4- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrro [3,2-b ] pyridin-5-yl) butanamide (6-4) (60 mg,0.096mmol, 70.86%). LC-MS (ESI) [ m+h ] + = 592.4.
In a fifth step, the reaction 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrrolio [3,2-b ] pyridin-5-yl) butanamide (6-4) (49 mg,0.083 mmol), tetrabutylammonium fluoride (0.830 mL,0.830 mmol) and the solvent tetrahydrofuran (3 mL) were added to a 25mL round bottom flask, the reaction was stirred overnight at 50℃and monitored by LC-MS. After the reaction, 15mL of ethyl acetate was added for dilution, and the mixture was washed with 20mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by prep. plate (developer: petroleum ether: ethyl acetate=2/1) to give 4- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrrolo [3,2-b ] pyridin-5-yl) butanamide as a white solid (6)(35.07mg,0.071mmol,85.86％).1H NMR(400MHz,CDCl3)δ8.36(s,1H),8.18(d,J＝8.9Hz,1H),7.67(d,J＝8.9Hz,1H),7.25(d,J＝3.2Hz,1H),6.48(d,J＝3.1Hz,1H),3.82(s,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.45–2.36(m,2H),2.17(dt,J＝16.3,7.1Hz,3H),2.04–1.86(m,4H),1.79–1.64(m,6H),1.57(dd,J＝13.2,3.7Hz,1H),1.37(ddd,J＝25.9,13.0,3.7Hz,2H),1.27(d,J＝11.9Hz,1H),1.12–1.06(m,1H),1.03(d,J＝4.3Hz,1H),0.97(d,J＝5.8Hz,3H),0.90(dd,J＝12.5,3.8Hz,1H),0.86–0.83(m,1H),0.82(d,J＝7.6Hz,4H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ132.48,132.39,127.79,127.64,119.04,108.04,100.44,54.28,53.97,38.83,37.80,37.73,36.96,36.52,33.56,33.33,32.11,28.46,28.06,27.66,24.30,21.92,21.85,19.64,15.38,14.30.LC-MS(ESI)[M+H]+＝478.3.
Example 7
Preparation of the Compound N- (benzo [2,1-c ] [1,2,5] thiadiazepin-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
The compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108 mmol) was added to a reaction flask containing methylene chloride (2 mL), oxalyl chloride (0.018 mL,0.216 mmol) and N, N-dimethylformamide (0.05 mL) were added and stirred at room temperature (20 ℃ C.) for 0.5hr, the reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a colorless oil, and sodium hydride (8.64 mg,0.216 mmol) and benzo [ c ] [1,2,5] thiadiazepine-5-amine (19.60 mg,0.130 mmol) were further stirred for 18hr; the reaction liquid is monitored by LC-MS to have no raw material residue; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product N- (benzo [2,1-c ] [1,2,5] thiadiazepin-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (7) (4 mg,0.008mmol, 7.46%) as a white solid
1H NMR(400MHz,CDCl3)δ8.42(s,1H),7.93(d,J＝9.3Hz,1H),7.59(dd,J＝9.4,2.1Hz,1H),7.29(d,J＝11.6Hz,1H),5.35(m,1H),3.22(dd,J＝11.5,4.6Hz,1H),2.41(s,2H),2.20(m,3H),2.00(d,J＝8.9Hz,2H),1.90(d,J＝14.8Hz,1H),1.76(m,1H),1.69(m,2H),1.63(s,3H),1.40(m,2H),1.30(s,2H),1.26(s,3H),1.05(m,2H),0.97(s,2H),0.88(s,2H),0.82(s,3H),0.79(s,2H).LC-MS(ESI)[M+H]+＝478.30.
Example 8
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] butanamide
In a first step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (60 mg,0.130 mmol), 2- (1-methylindol-5-yl) ethan-1-amine (27.11 mg,0.156 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (73.95 mg,0.194 mmol), N, N-diisopropylethylamine (0.064 mL,0.389 mmol) and solvent N, N-dimethylformamide (1.5 mL) were added to a 5mL round bottom flask, and the reaction solution was stirred at room temperature and monitored by LC-MS. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: petroleum ether: ethyl acetate=3/1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] butanamide (8-1) as a pale yellow solid (50 mg,0.075mmol, 57.89%). LC-MS (ESI) [ m+h ] + =619.4.
In a second step, the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] butanamide (8-1) (50 mg,0.081 mmol), tetrabutylammonium fluoride (211.20 mg, 0.806 mmol) and solvent tetrahydrofuran (2 mL) were charged in a 25mL round bottom flask, and the reaction was stirred at 50℃for 28 hours, as monitored by LC-MS. After the reaction was completed, 15mL of ethyl acetate was added for dilution, and the mixture was washed 3 times with 20mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. Purification of the crude product by preparative plate (dichloromethane/methanol=10/1) gives 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (1-methylindol-5-yl) ethyl ] butyramide as a white solid (8)(18.08mg,0.033mmol,40.78％).1H NMR(400MHz,DMSO)δ7.80(t,J＝5.5Hz,1H),7.32(d,J＝8.6Hz,2H),7.26(d,J＝3.0Hz,1H),6.98(dd,J＝8.3,1.4Hz,1H),6.32(d,J＝2.8Hz,1H),4.31(d,J＝5.1Hz,1H),3.75(s,3H),3.26(dd,J＝14.1,6.4Hz,2H),3.04–2.96(m,1H),2.75(s,2H),2.01(s,9H),1.53(s,10H),1.45–1.20(m,4H),1.01–0.96(m,1H),0.88(s,3H),0.76(s,3H),0.69(s,3H).13C NMR(101MHz,DMSO)δ172.33,135.69,133.19,130.26,130.04,128.66,126.75,122.59,120.28,109.85,100.30,77.39,54.37,53.99,41.44,38.93,38.09,37.05,36.52,35.95,33.51,32.90,32.20,28.82,28.64,27.90,24.90,22.09,21.92,19.78,16.27,14.57.LC-MS(ESI)[M+H]+＝505.3.
Example 9
Preparation of the Compound 4- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [ (1-methylindol-5-yl) methyl ] butanamide
The first step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (80 mg,0.173mmol,1.0 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (2 mL), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (78.87 mg,0.207mmol,1.2 eq) and N, N-diisopropylethylamine (0.057 mL,0.346mmol,2.0 eq) were added, and stirring was continued at room temperature (18 ℃) for 0.5 hours, followed by (1-methylindol-5-methanamine) (0.33 mg, 1.2 eq); TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction showed the disappearance of starting material. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give the crude product which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1) to give 4- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (1-methylindol-5-yl) methyl ] butyramide (9-1) (52.6 mg,0.087mmol, 50.07%) as a white solid.
The second step compound 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (1-methylindol-5-yl) methyl ] butanamide (13-1) (50 mg,0.096mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.962 mL,0.962mmol,10 eq) was added thereto, followed by heating to 50℃and stirring for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitoring the reaction showed substantial disappearance of starting material; the reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a brown oil, which was added with water (20 mL), extracted with ethyl acetate (20 ml×3), the organic phases were combined, washed with water (30 ml×2), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:2) to give the product 4- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -N- [ (1-methylindol-5-yl) methyl ] butyramide (9) (17.42 mg,0.038mmol, 39.44%) as a white solid ;1H NMR(400MHz,DMSO)δ8.21(t,J＝5.8Hz,1H),7.44–7.25(m,3H),7.05(d,J＝8.4,1.4Hz,1H),6.35(d,J＝3.0Hz,1H),4.31(t,J＝5.0Hz,3H),3.76(s,3H),3.04–2.95(m,1H),2.09(t,J＝7.1Hz,3H),1.99–1.85(m,J＝33.7,9.5Hz,4H),1.66–1.57(m,3H),1.53(d,J＝7.2Hz,3H),1.50–1.37(m,3H),1.32–1.21(m,2H),1.04–0.91(m,J＝20.2,12.3,7.8Hz,2H),0.88(d,J＝7.4Hz,3H),0.75(s,4H),0.69(d,J＝8.8Hz,3H).13C NMR(101MHz,DMSO)δ172.25,136.01,133.19,130.64,130.29,128.38,126.80,121.59,119.49,109.85,100.51,77.38,54.37,53.98,42.96,38.92,38.11,37.05,36.52,35.95,33.49,32.95,32.21,28.83,28.63,27.90,24.96,22.06,21.91,19.78,16.26,14.56.LC-MS(ESI)[M+H]+＝491.73.
Example 10
Preparation of the Compound N- (1, 3-dimethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
In the first step, the compound 5-nitro-1H-indole-3-carbaldehyde (500 mg,2.629mmol,1.0 eq) was dissolved in DMF (10 mL), then nitrogen was replaced, cooled to 0℃and then sodium hydrogen (210.35 mg, 5.299 mmol,2.0 eq) was added thereto, after stirring for 30 minutes, methyl iodide (0.164 mL,2.629mmol,1.0 eq) was added thereto, followed by stirring at room temperature overnight. LC-MS detection, after the reaction was completed, saturated sodium bicarbonate was added thereto, then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated saline, then dried over anhydrous sodium sulfate, and spin-dried to give a crude product, which was purified by flash chromatography (dichloromethane/methanol=1:0 to 10:1) to give 1-methyl-5-nitroindole-3-carbaldehyde (10-1) (271 mg, 50.48%) as a pale yellow solid. LC-MS (ESI) [ m+1] + =205.1
In the second step, the compound 1-methyl-5-nitroindole-3-carbaldehyde (10-1) (100 mg,0.490mmol,1.0 eq) was dissolved in ethanol (10 mL), then Pd/C (52.12 mg,0.490mmol,10.0 eq) was added thereto, hydrogen was replaced, and the temperature was raised to 40℃and stirred for 3 hours. LC-MS detection, when the reaction was completed, filtration and spin-drying gave 1, 3-dimethylindol-5-amine (10-2) (48 mg, yield: 61.17%) as an oily compound. LC-MS (ESI) [ m+1] + =161.1
In the third step, 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (30 mg,0.086mmol,1.0 eq), HATU (49.09 mg,0.129mmol,1.5 eq) and 1, 3-dimethylindol-5-amine (16.55 mg,0.103mmol,1.2 eq) were dissolved in DMF (3 mL), then N, N-diisopropylethylamine (0.036 mL,0.215mmol,2.5 eq) was added thereto and stirred overnight at room temperature. LC-MS monitored the reaction, after which water was added to quench it after the reaction was completed, then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, dried by spin-drying, and then purified by preparative plate separation (petroleum ether: ethyl acetate=2:1) to give the solid compound N- (1, 3-dimethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (31 mg, yield: 69.72%) as a white solid .1H NMR(400MHz,CDCl3)δ7.77(s,1H),7.26(s,1H),7.20(d,J＝1.7Hz,3H),6.81(s,1H),3.73(d,J＝18.2Hz,3H),3.22(dd,J＝11.3,4.3Hz,1H),2.35(dd,J＝15.6,8.5Hz,2H),2.29(d,J＝7.1Hz,3H),2.17(t,J＝11.2Hz,3H),1.99(d,J＝7.3Hz,2H),1.89(dd,J＝17.6,11.8Hz,2H),1.80–1.68(m,4H),1.61(d,J＝15.6Hz,5H),1.41–1.32(m,2H),1.26(s,1H),1.12–1.03(m,2H).13C NMR(101MHz,CDCl3)δ171.17,134.52,132.54,129.57,128.67,127.67,127.45,115.80,111.16,110.23,109.10,79.13,54.30,53.98,38.83,37.85,37.57,36.97,36.54,33.60,32.62,32.14,28.56,28.06,27.66,24.55,21.94,21.88,19.69,15.38,14.31,9.57.LC-MS(ESI)[M+1]+＝491.3.
Example 11
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [2- (pyrazin-2-yl) ethyl ] indol-5-yl } butanamide
The first step of the compound 5-nitro-1H-indole (750 mg,4.625 mmol) and 2-vinyl pyrazine (490.89 mg,4.625mmol,1.0 eq) were added to a reaction flask containing dimethyl sulfoxide (8 mL), potassium hydroxide (519.06 mg,9.251mmol,2.0 eq) and water (4 mL) were added, and after heating to 120deg.C and stirring for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitoring the reaction showed starting material remained; the reaction mixture was taken up in water (20 mL) and extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried under reduced pressure (water pump, 45 ℃ C.) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give the product 5-nitro-1- [2- (pyrazin-2-yl) ethyl ] indole (11-1) (500 mg,1.677mmol, 36.26%) as a yellow solid .1H NMR(400MHz,CDCl3)δ8.56(d,J＝2.2Hz,2H),8.44(d,J＝2.5Hz,1H),8.19(d,J＝1.3Hz,1H),8.08(dd,J＝9.1,2.2Hz,1H),7.30(d,J＝9.1Hz,1H),7.13(d,J＝3.3Hz,1H),6.63(d,J＝3.2Hz,1H),4.66(t,J＝6.9Hz,2H),3.34(t,J＝6.9Hz,2H).
The second step compound, 5-nitro-1- [2- (pyrazin-2-yl) ethyl ] indole (11-1) (50 mg,0.186mmol,1.0 eq), was added to a reaction flask containing methanol (2 mL), and after addition of palladium on carbon 10% (10 mg,0.009mmol,0.05 eq), the mixture was stirred under hydrogen atmosphere (15 psi) at room temperature 25℃for 16hr. LC-MS (CL 210743-275-R1) monitors no starting material remaining; the reaction mixture was filtered through celite, the filter cake was washed with methanol (10 mL), the filtrate was dried under reduced pressure (water pump, 40 ℃ C.), and the oil pump was pulled dry to give the product 1- [2- (pyrazin-2-yl) ethyl ] indol-5-amine (11-2) (48 mg,0.181mmol, 97.28%) as a tan oil .1H NMR(400MHz,CDCl3)δ8.57–8.52(m,1H),8.41(d,J＝2.5Hz,1H),8.16(s,1H),7.12(d,J＝8.6Hz,1H),6.91(d,J＝1.9Hz,1H),6.84(d,J＝3.1Hz,1H),6.66(dd,J＝8.6,2.1Hz,1H),6.23(d,J＝3.0Hz,1H),4.50(t,J＝7.0Hz,2H),3.28(t,J＝7.0Hz,2H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (40 mg,0.086mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (32.87 mg,0.086mmol,1.0 eq) and N, N-Diisopropylethylamine (DIEA) (0.014 mL,0.172mmol,2.0 eq) were added and then stirred at room temperature (25 ℃) for 0.5hr, followed by 1- [2- (pyrazin-2-yl) ethyl ] indol-5-amine (11-2) (24.72 mg,0.104 eq, 1.1 hr) at room temperature; TLC (dichloromethane: methanol=20:1) (petroleum ether: ethyl acetate=1:1) monitoring the reaction showed the disappearance of starting material; the reaction mixture was added with ethyl acetate (10 mL) and water (20 mL), extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with saturated aqueous saline (20 mL x 3), dried under reduced pressure (water pump, 40 ℃) to give crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {1- [2- (pyrazin-2-yl) ethyl ] indol-5-yl } butanamide (11-3) (40 mg,0.053mmol, 60.98%) as white solid .1H NMR(400MHz,CDCl3)δ8.57(s,1H),8.45(d,J＝2.2Hz,1H),8.15(s,1H),7.75(s,1H),7.24(d,J＝4.9Hz,1H),7.11(s,1H),6.92(d,J＝3.0Hz,1H),6.38(d,J＝2.9Hz,1H),4.58(t,J＝6.9Hz,2H),3.33(t,J＝6.8Hz,2H),3.18(dd,J＝11.2,4.5Hz,1H),2.35(dd,J＝15.2,8.8Hz,2H),2.17(d,J＝8.8Hz,3H),1.96(dd,J＝42.4,22.9Hz,6H),1.62(s,3H),1.52(d,J＝9.4Hz,3H),1.38(d,J＝4.6Hz,1H),1.03(dd,J＝23.2,12.7Hz,3H),0.91–0.86(m,15H),0.81(s,3H),0.80(s,1H),0.75(s,3H),0.03(d,J＝2.7Hz,6H).
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [2- (pyrazin-2-yl) ethyl ] indol-5-yl } butanamide (11-3) (40 mg,0.059mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.586 mL,0.586mmol,10 eq), the mixture was stirred at 50℃for 16hr; the reaction was monitored by TLC (petroleum ether: ethyl acetate=1:2) to show a small amount of starting material remained; the reaction mixture was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was dissolved in ethyl acetate (20 mL), washed with water (15 mL x 3), dried under reduced pressure, and purified by prep-TLC (petroleum ether: ethyl acetate=1:2) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {1- [2- (pyrazin-2-yl) ethyl ] indol-5-yl } butanamide (11) (15.87 mg,0.027mmol, 45.81%) as a white solid .1H NMR(400MHz,CDCl3)δ8.55(s,1H),8.43(s,1H),8.15(s,1H),7.76(s,1H),7.24(s,1H),7.12(s,1H),6.92(d,J＝3.0Hz,1H),6.38(d,J＝3.0Hz,1H),4.56(t,J＝6.9Hz,2H),3.29(t,J＝6.9Hz,2H),3.22(dd,J＝11.4,4.6Hz,1H),2.44–2.28(m,2H),2.17(m,3H),1.98(m,2H),1.95–1.84(m,2H),1.84–1.73(m,2H),1.72(m,2H),1.66(m,2H),1.63(s,3H),1.51(m,1H),1.38(m,1H),1.06(m,1H),1.00(m,1H),0.97(s,3H),0.94–0.89(m,1H),0.87(m,1H),0.85(m,1H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ171.14,153.99,144.25,144.24,144.22,132.52,130.33,128.76,128.47,127.71,116.06,112.99,109.24,101.71,79.13,77.34,77.22,77.02,76.70,54.31,53.99,45.54,38.84,37.85,37.57,36.98,36.54,35.97,33.59,32.14,28.55,28.07,27.66,24.53,21.94,21.88,19.69,15.38,14.31.LC-MS(ESI)[M+1]+＝569.5.
Example 12
Preparation of the Compound N- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1-methylindole-5-carboxamide
The first step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propan-1-ol (II-1) (6 g,13.800mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (100 mL), p-toluenesulfonyl chloride (4.74 g, 24.84mmol, 1.8 eq), 4-dimethylaminopyridine (1.35 g,11.040mmol,0.8 eq) and N, N-diisopropylethylamine (4.561 mL,27.600mmol,2.0 eq) were added, and then stirred at 40℃for 4 hours; TLC (petroleum ether: ethyl acetate=2:1) monitoring the reaction showed no starting material remaining; dichloromethane (30 mL) was added to the reaction solution, which was washed with saturated aqueous sodium hydrogencarbonate (100 mL) and water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 19:1) to give 4-methylbenzenesulfonic acid-3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl ester (12-1) (5 g,7.640mmol, 55.37%) as a white solid .1H NMR(400MHz,CDCl3)δ7.80(d,J＝8.3Hz,2H),7.35(d,J＝8.0Hz,2H),4.01(dd,J＝6.4,3.3Hz,2H),3.17(dd,J＝11.1,4.7Hz,1H),2.45(s,3H),2.13–1.98(m,4H),1.85(d,J＝11.3Hz,3H),1.77–1.66(m,3H),1.65–1.57(m,4H),1.53(s,3H),1.37–1.29(m,2H),1.03–0.93(m,3H),0.89(s,12H),0.77(s,3H),0.74(s,3H),0.03(d,J＝2.7Hz,6H).
The second step compound 4-methylbenzenesulfonic acid-3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (12-1) (200 mg,0.340mmol,1.0 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (5 mL), phthalimide methyl salt (76 mg,0.410mmol,1.2 eq) and tetrabutylammonium iodide (26 mg,0.070mmol,0.2 eq) was added, followed by stirring at 100℃for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitoring the reaction showed no starting material remaining; ethyl acetate (30 mL) was added to the reaction solution, water (100 mL) was washed, and then the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give the product 2- {3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } isoindole-1, 3-dione (12-2) (120 mg,0.213mmol, 62.65%) as a yellow oil .1H NMR(400MHz,CDCl3)δ7.81(s,1H),7.69(s,1H),3.67–3.59(m,1H),3.14(dd,J＝11.1,4.6Hz,1H),2.08(tdd,J＝15.4,10.1,5.3Hz,1H),1.93(t,J＝11.2Hz,1H),1.86–1.72(m,1H),1.65–1.59(m,1H),1.55(d,J＝7.5Hz,3H),1.40(s,1H),1.34–1.21(m,1H),0.96–0.89(m,1H),0.86(s,4H),0.84(s,2H),0.77(s,2H),0.71(s,2H),0.00(d,J＝2.7Hz,3H).
The third step compound 2- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } isoindole-1, 3-dione (12-2) (120 mg,0.213mmol,1 eq) was added to a reaction flask and then stirred at room temperature (80 ℃) for 12 hours after hydrazine hydrate (2 ml) and ethanol (0.5 ml) were added; TLC (dichloromethane: methanol=20:1) monitored the reaction showed the disappearance of starting material, stopped the reaction, filtered off the white solid and the mother liquor was collected after spin-drying to give crude 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propanamide (12-3) (65.4 mg,0.151mmol, 70.89%) as a white solid, which was directly thrown back.
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylhydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propanamide (12-3) (65.4 mg,0.151mmol,1 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (2 mL), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (69.00 mg,0.267mmol,1.2 eq) and N, N-diisopropylethylamine (0.051mL, 0.446mmol, 2.0.0 eq) were added and stirred at room temperature (25 ℃ C.) for 0.5 hours, and stirred for a further 1-5.151mg (1 mmol) of indole; TLC (dichloromethane: methanol=20:1) monitoring the reaction showed the starting material disappeared; the reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) to give the crude product which was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give the product N- {3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } -1-methylindole-5-carboxamide (12-4) (47.8 mg,0.081mmol, 53.64%) as a yellow oil, which was directly added to the latter.
The fifth step compound N- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthryl ] propyl } -1-methylindole-5-carboxamide (12-4) (47.8 mg,0.081mmol,1 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.81ml, 0.81mmol, 10eq, 1M) was added thereto and stirred at 50℃for 16 hours; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed a small amount of starting material remaining; the reaction solution was dried under reduced pressure (water pump, 45 ℃) and added with ethyl acetate (20 mL) and water (20 mL) to dissolve, extracted with ethyl acetate (15 ml×3), the organic phases were combined, washed with water (20 ml×4) and dried under reduced pressure (water pump, 45 ℃) to give crude product, which was separated and purified by column chromatography (dichloromethane: methanol=10:1) to give product N- {3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } -1-methylindole-5-carboxamide (12) (32.05 mg,0.059mmol, 34.13%) as white solid .1H NMR(400MHz,CDCl3)δ8.05(s,1H),7.67(d,J＝8.6Hz,1H),7.34(d,J＝8.6Hz,1H),7.11(d,J＝3.1Hz,1H),6.55(d,J＝3.0Hz,1H),6.23(s,1H),3.82(s,3H),3.52–3.43(m,2H),3.21(dd,J＝11.5,4.6Hz,1H),2.19(ddd,J＝16.2,13.2,5.6Hz,3H),2.03–1.91(m,3H),1.79–1.73(m,2H),1.72–1.65(m,3H),1.62(d,J＝7.7Hz,7H),1.58(d,J＝3.5Hz,1H),1.41–1.34(m,1H),1.05(dd,J＝10.7,5.6Hz,1H),0.96(s,3H),0.89–0.82(m,2H),0.79(d,J＝7.1Hz,6H).13C NMR(101MHz,CDCl3)δ168.56,138.25,132.58,130.19,128.02,127.57,126.22,120.58,120.12,109.10,102.12,79.10,54.27,53.93,40.32,38.82,37.87,36.94,36.52,33.57,33.03,32.16,28.49,28.05,27.64,26.60,21.92,21.82,19.65,15.37,14.27.LC-MS(ESI)[M+H]+＝477.34.
Example 13
Preparation of the Compound N- (1, 4-dimethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
In the first step, the compound 4-methyl-5-nitro-1H-indole (500 mg,2.84mmol,1.0 eq) was dissolved in N, N-dimethylformamide (10 mL), then potassium carbonate (902.09 mg,6.53mmol,2.3 eq) and methyl iodide (0.88 mL,14.19mmol,5.0 eq) were added thereto, nitrogen was replaced, stirring was carried out at room temperature overnight, after completion of the reaction, water quenching was added, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine (50 mL x 2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give 1, 4-dimethyl-5-nitroindole (13-1) (479 mg,2.52mmol, 88.74%) as a yellow solid. LC-MS (ESI) [ m+h ] + = 191.1
In the second step, the compound 1, 4-dimethyl-5-nitroindole (13-1) (100 mg,0.526mmol,1.0 eq) was dissolved in methanol (10 mL), then palladium (palladium on charcoal) (55.95 mg,0.526mmol,1.0 eq) was added thereto to displace hydrogen, then stirred at room temperature for 2 hours, TLC and LC-MS detection were performed, and after completion of the reaction, filtration and then spin-drying gave 1, 4-dimethylindol-5-amine (13-2) (80 mg,0.50mmol, 94.97%) as an oily compound. LC-MS (ESI) [ m+h ] + =161.1
In a third step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butyryl chloride (14-1) (62 mg,0.129mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), then 1, 4-dimethylindol-5-amine (13-2) (29.96 mg,0.187mmol,1.5 eq) and triethylamine (0.052 mL,0.374mmol,3.0 eq) were added thereto, and finally 4-dimethylaminopyridine (7.62 mg, 0.mmol, 0.5 eq) was added and stirred at room temperature, then allowed to warm up to 50℃for 3h, detection, after completion of the reaction, water was added thereto, and then sodium acetate was dried overnight (50 mL) was extracted with a dry plate, and the crude product was prepared by filtration, dry-phase was dried, dried and dried over a sodium sulfate (dry plate). Ethyl acetate = 2:1) to give N- (1, 4-dimethylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (13-3) (50 mg,0.083mmol, 66.29%) as an oily compound .1H NMR(400MHz,CDCl3)δ7.32(d,J＝8.6Hz,1H),7.13–6.80(m,3H),6.58–6.32(m,1H),3.77(t,J＝14.5Hz,3H),3.18(dd,J＝11.0,4.6Hz,1H),2.59–2.31(m,5H),2.17(d,J＝6.9Hz,2H),1.96(m,4H),1.81–1.43(m,13H),1.32(m,2H),1.15–0.93(m,3H),0.88(d,J＝4.9Hz,9H),0.86(s,1H),0.81(d,J＝7.0Hz,3H),0.76–0.70(m,3H),0.03(d,J＝2.6Hz,6H).
The fourth step was to dissolve the compound N- (1, 4-dimethylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (9-3) (50 mg,0.083mmol,1.0 eq) in tetrahydrofuran (5 mL), then to which tetrabutylammonium fluoride (1M in tetrahydrofuran) (0.83 mL,0.826mmol,10.0 eq) was added, then to replace nitrogen, then to stir at 50 degrees overnight, LC-MS and TLC were quenched, water was then added after completion of the reaction, and then extracted with ethyl acetate (50 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, and spun dry to give the crude petroleum ether, prepared plate (petroleum ether: ethyl acetate=2:1) to give an oily compound, which was lyophilized to give N- (1, 4-dimethylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (13) (22.75 mg,0.045mmol, 53.85%) as a white solid .1H NMR(400MHz,CDCl3)δ7.31(d,J＝8.6Hz,1H),7.18–7.10(m,1H),7.03(t,J＝4.2Hz,1H),6.98(s,1H),6.50(dd,J＝18.1,3.0Hz,1H),3.79(d,J＝16.2Hz,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.55–2.26(m,5H),2.25–2.14(m,2H),2.01(d,J＝4.7Hz,2H),1.95–1.87(m,2H),1.81–1.69(m,3H),1.64(s,3H),1.59(dd,J＝14.3,10.4Hz,3H),1.50–1.23(m,5H),1.06(dd,J＝12.9,4.5Hz,1H),0.96(d,J＝7.3Hz,3H),0.91–0.86(m,1H),0.81(d,J＝5.9Hz,3H),0.78(d,J＝6.8Hz,3H).13C NMR(101MHz,CDCl3)δ171.82,134.83,132.59,128.87,127.68,126.69,124.28,120.24,107.12,99.91,79.12,77.35,77.23,77.03,76.71,54.30,53.99,38.83,37.92,37.20,36.97,36.54,33.58,33.01,32.14,28.74,28.06,27.65,24.81,21.94,21.87,19.69,15.38,14.31,14.09.LC-MS(ESI)[M+H]+＝491.3.
Example 14
Preparation of the Compound N- [4- (dimethylamino) phenyl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
In a first step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (120 mg, 0.299 mmol,1.0 eq) was dissolved in methylene chloride (5 mL), then N, N-dimethylformamide (0.020mL, 0.299 mmol,1.0 eq) and oxalyl chloride (0.044 mL, 0.719 mmol,2.0 eq) were added thereto, then stirred at room temperature for one hour, TLC was performed (one drop of methanol TLC was used), and after completion of the reaction, it was spin-dried to give 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butyryl chloride (14-1) (120 mg, 0.219 mmol, 96.17%) as an oily compound.
In a second step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butyryl chloride (14-1) (62 mg,0.129mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), and then a solution of 4- (dimethylamino) aniline (26.32 mg,0.193mmol,1.5 eq) and triethylamine (0.054 mL,0.387mmol,3.0 eq) in tetrahydrofuran was added thereto. After stirring at room temperature for two hours, LC-MS detection, water was added thereto, extraction was performed with ethyl acetate, and the organic phase was dried over sodium sulfate, filtered and spin-dried to give a crude product, which was then isolated and purified (petroleum ether: ethyl acetate=1:1) via preparative plate to give N- [4- (dimethylamino) phenyl ] -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (14-2) (50 mg,0.086mmol, 66.8%) as a white solid. LC-MS (ESI) [ m+h ] + =581.4
In a third step, the compound N- [4- (dimethylamino) phenyl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (14-2) (50 mg,0.086mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), then tetrabutylammonium fluoride (225.03 mg,0.861mmol,10.0 eq) was added thereto, and the reaction was allowed to stand at 50℃overnight, and LC-MS and TLCTLC were examined, after completion of the reaction of the starting material, quenched with water, then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated sodium chloride (50 mL), then dried over anhydrous sodium sulfate, filtered, and spun dry to give a crude product, and then the plate (petroleum ether was prepared: ethyl acetate=1:1) to give an oily compound, and freeze-drying to give N- [4- (dimethylamino) phenyl ] -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (14) (26 mg,0.05mmol, 61.49%) as a white solid .1H NMR(400MHz,CDCl3)δ7.34(d,J＝8.9Hz,2H),7.00(s,1H),6.70(d,J＝7.8Hz,2H),3.22(dd,J＝11.4,4.5Hz,1H),2.97–2.79(m,6H),2.30(dd,J＝15.8,8.4Hz,2H),2.20–2.09(m,3H),2.04–1.90(m,3H),1.89–1.73(m,3H),1.71–1.63(m,5H),1.61(s,3H),1.41–1.32(m,2H),1.09–1.00(m,2H),0.96(s,3H),0.89–0.83(m,2H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ170.95,132.50,127.67,121.82,113.15,79.12,77.34,77.03,76.71,54.29,53.96,40.98,38.83,37.83,37.43,36.97,36.52,33.58,32.11,28.53,28.06,27.65,24.50,21.92,21.86,19.67,15.38,14.30.LC-MS(ESI)[M+H]+＝467.3.
Example 15
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-methyl-N- (1-methylindol-5-yl) butanamide
The first step compound 5-bromo-1-methylindole (500 mg,2.380mmol,1.0 eq) was added to a reaction flask containing DMF (10 mL), methylamine (7.140 mL,14.281mmol,6.0 eq) was added, strontium carbonate (1551.01 mg,4.760mmol,2.0 eq) and tBuXPhos-Pd-G3 (189.32 mg,0.238mmol,0.1 eq) were stirred at room temperature (80 ℃) for 18hr and the reaction solution was monitored by LC-MS. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give the product 1-methyl-5- (methylamino) indole (15-1) (200 mg,0.999mmol, 41.96%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 161.20.
The second step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (30 mg,0.065mmol,1.0 eq) was added to a reaction flask containing methylene chloride (2 mL), oxalyl chloride (0.01 mL,0.130mmol,2.0 eq) and N, N-dimethylformamide (0.05 mL) were added and stirred at room temperature (20 ℃ C.) for 0.5hr, the reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a colorless oil, and THF (2 mL), N, N-diisopropylethylamine (0.032 mL,0.194mmol, 3.07 eq) and 1-methyl-5- (methylamino) indole (15.460 mg,1.0 mmol, 18 hr) were further added and stirred; the reaction liquid is monitored by LC-MS, no raw material remains, and products are generated (Ms+1= 492.4); the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1=1:1) to give the product 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-methyl-N- (1-methylindol-5-yl) butanamide (15-2) (25 mg,0.037mmol, 57.22%) as a pale yellow oil 1H NMR(400MHz,CDCl3)δ7.38(d,J＝1.7Hz,1H),7.30(d,J＝8.6Hz,1H),7.10(d,J＝3.0Hz,1H),6.98(dd,J＝8.6,1.8Hz,1H),6.46(d,J＝3.0Hz,1H),3.80(s,3H),3.27(s,3H),3.13(dd,J＝11.0,4.5Hz,1H),2.03(m,2H),1.98(d,J＝3.7Hz,1H),1.86(dd,J＝9.3,7.0Hz,2H),1.75(dd,J＝16.4,4.8Hz,2H),1.63(m,2H),1.55(m,3H),1.45(s,3H),1.31(s,1H),1.22(dd,J＝18.6,7.5Hz,5H),0.90(m,2H),0.86(s,9H),0.83(s,3H),0.75(dd,J＝8.0,4.7Hz,2H),0.70(s,3H),0.68(s,2H),0.00(d,J＝2.5Hz,6H).LC-MS(ESI)[M+H]+＝474.30
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-methyl-N- (1-methylindol-5-yl) butanamide (15-2) (25 mg,0.041mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (2 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (0.413 mL,0.413mmol,10.0 eq), the mixture was warmed to 50℃and stirred for 18hr; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed that the starting material was essentially reacted; the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-methyl-N- (1-methylindol-5-yl) butanamide (15) (15 mg,0.028mmol, 66.57%) as a white solid .1H NMR(400MHz,CDCl3)δ7.41(d,J＝1.8Hz,1H),7.33(d,J＝8.6Hz,1H),7.13(d,J＝3.1Hz,1H),7.01(dd,J＝8.6,1.9Hz,1H),6.50(d,J＝3.0Hz,1H),3.83(s,3H),3.30(s,3H),3.20(dd,J＝11.5,4.6Hz,1H),2.05(dd,J＝10.1,4.2Hz,2H),1.89(dd,J＝9.5,7.0Hz,2H),1.80(s,1H),1.70(m,2H),1.62(m,5H),1.47(s,3H),1.33(s,1H),1.25(s,6H),0.95(s,3H),0.85(m,2H),0.77(s,4H),0.72(s,3H).13C NMR(101MHz,CDCl3)δ173.95,136.51,135.63,132.92,130.29,128.88,127.10,120.85,119.40,110.07,101.29,79.12,54.26,53.91,38.80,37.90,36.94,36.46,34.30,33.47,33.08,32.01,31.44,30.19,29.71,29.37,28.73,28.06,27.64,24.63,21.87,21.78,19.49,15.40,14.21,14.13.LC-MS(ESI)[M+H]+＝491.35.
Example 16
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] propanamide
The first step compound (3E) -hex-3-enedioic acid (1 g,6.94mmol,1.0 eq) was added to a reaction flask containing methanol (10 mL), concentrated sulfuric acid (1 mL) was added and stirred at 80℃for 12 hours; TLC (petroleum ether: ethyl acetate=20:1) monitored the reaction. Ethyl acetate (20 mL) was added to the reaction solution, which was washed with saturated aqueous sodium bicarbonate (20 mL x 2), and dried by spin drying under reduced pressure (water pump, 40 ℃) to give methyl (3E) -6-methoxy-6-oxohex-3-enoate (16-1) (1 g, 5.178 mmol, 83.71%) as a colorless oily liquid, which was directly added to the crude product.
The second step of the compound (3E) -6-methoxy-6-oxohex-3-enoic acid methyl ester (16-1) (1 g, 5.806 mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (20 mL), and lithium aluminum hydride (0.32 g,8.42mmol,1.3 eq) was added after hydrogen was replaced and stirred for 2 hours; TLC (petroleum ether: ethyl acetate=1:1) showed that the reaction was complete. Sodium sulfate decahydrate is added until bubbling is avoided, quenching is completed, crude product obtained after spinning is separated and purified by column chromatography (dichloromethane: methanol=10:1) to obtain (3E) -hex-3-ene-1, 6-diol (16-2) (560 mg, 4.8231 mmol, 83.01%) as colorless oil .1H NMR(400MHz,CDCl3)δ5.48–5.36(m,1H),4.45(t,J＝5.3Hz,1H),3.42–3.35(m,2H),2.17–2.05(m,2H).
The third step compound (3E) -hex-3-ene-1, 6-diol (16-2) (560 mg, 4.823 mmol,1.0 eq) was added to a reaction flask containing dichloromethane (10 mL), triphenylphosphine (1.9 g,7.23mmol,1.5 eq) was added, and carbon tetrabromide (2.4 g,7.23mmol,1.5 eq) was stirred at room temperature (18 ℃ C.) for 16 hours; TLC (petroleum ether: ethyl acetate=20:1) monitored complete starting material. The reaction solution was dried under reduced pressure (water pump, 50 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product (3E) -1, 6-dibromohex-3-ene (16-3) (250 mg,1.033mmol, 21.4%) as a yellow oily liquid; the new point is thrown directly backwards.
The fourth step compound (3E) -1, 6-dibromohex-3-ene (16-3) (250 mg,1.033mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (5 mL), after nitrogen substitution, cooled to-78 ℃, lithium diisopropylamide (110.69 mg,1.24mmol,1.2eq, 1M) was slowly added dropwise, after stirring for 0.5 hour, 2-methylpyridine (96.23 mg,1.033mmol,1eq, 1M) was added dropwise and allowed to react for two hours at room temperature, and TLC (petroleum ether: ethyl acetate=5:1) monitored the reaction. The reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give the product 2- [ (4E) -7-bromohept-4-enyl ] pyridine (16-4) (100 mg,0.393mmol, 38.08%) as a colorless oil. LC-MS (ESI) [ m+h ] + =254.
The fifth step compound 2- [ (4E) -7-Bromohept-4-enyl ] pyridine (16-4) (100 mg,0.393mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (3 mL), and after adding potassium phthalimide (146 mg,0.786mmol,2.0 eq), it was stirred at room temperature for 16 hours. LC-MS monitored the reaction. The reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give 2- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] isoindole-1, 3-dione (16-5) (100 mg,0.312mmol, 79%) as a white solid. LC-MS (ESI) [ m+h ] + =321
The sixth step compound, 2- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] isoindole-1, 3-dione (146-5) (100 mg,0.312mmol,1.0 eq) was added to a reaction flask containing ethanol (3 mL), and hydrazine hydrate (0.045 mL,0.744mmol,2.4 eq) was added and then heated to 80℃and stirred for 16hr. LC-MS monitored the reaction. The reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) and then was substituted with oil pump to give the product (3E) -7- (pyridin-2-yl) hept-3-en-1-amine (16-6) (40 mg,0.210mmol, 67%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =191
The seventh step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (37.46 mg,0.112mmol,1.0 eq)) was added to a reaction flask containing N, N-dimethylformamide (2 mL), and (7-azobenzotriazole) -N, N, N\' -tetramethylurea hexafluorophosphate (85.17 mg,0.224mmol,2 eq) and ethyl [ di (propan-2-yl) ] amine (0.037 mL,0.224mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5 hours, followed by (3E) -7- (pyridin-2-yl) hept-3-en-1-amine (16-6) (21.21.112 mg,1 mmol). TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] propionamide (16-7) (10 mg,0.0161mmol, 14%) as a pale yellow solid. The new point is thrown directly backwards.
The eighth step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] propanamide (16-7) (10 mg,0.0161mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (0.161 mL,0.161mmol,10 eq) was added and then warmed to 50℃and stirred for 16 hours. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ (3E) -7- (pyridin-2-yl) hept-3-enyl ] propionamide (16) (4.34 mg,0.0086mmol, 53%) as a white solid .1H NMR(400MHz,CDCl3)δ8.50(d,J＝4.2Hz,1H),7.60(s,1H),7.13(dd,J＝16.9,7.2Hz,2H),5.78(s,1H),5.50(dd,J＝14.7,7.5Hz,1H),5.37–5.29(m,1H),3.28(d,J＝4.2Hz,3H),2.80(t,J＝7.6Hz,2H),2.39(dt,J＝21.4,14.1Hz,2H),2.14(ddd,J＝48.3,13.1,5.8Hz,9H),1.83(dd,J＝22.9,15.4Hz,6H),1.59(s,3H),1.30(s,3H),1.10–1.00(m,2H),0.96(d,J＝7.3Hz,3H),0.85(dd,J＝13.6,7.7Hz,3H),0.80–0.76(m,6H).13C NMR(101MHz,CDCl3)δ172.95,161.95,149.10,136.36,132.59,131.93,128.34,127.62,122.78,121.04,79.08,54.27,53.93,38.87,38.81,37.97,37.46,36.93,36.57,35.92,35.69,33.52,32.68,32.08,31.95,29.32,28.05,27.62,27.22,25.26,21.87,21.79,19.52,15.37,14.28.LC-MS(ESI)[M+H]+＝507.
Example 17
Preparation of the Compound 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridazin-3-yl) heptyl ] propanamide
The first step compound 3-methylpyridazine (500.00 mg,5.312mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (20 mL), and after-78 ℃ addition of lithium diisopropylamide (3.187mL,6.74mmol 1.2eq), stirred at-78 ℃ for 30min; 1, 6-dibromohexane (1296.06 mg,5.312mmol,1.0 eq) was added and stirred at room temperature for 18h, and the reaction solution was monitored by TLC (petroleum ether: ethyl acetate=1:1) for the formation of a point of reduced polarity; the reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0to2:1) to give the product 3- (7-bromoheptyl) pyridazine (17-1) (450 mg,1.575mmol, 29.64%) as a colorless oily liquid LC-MS (ESI) [ m+h ] + = 257.10
The second step compound 3- (7-bromoheptyl) pyridazine (17-1) (314 mg,1.221mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (10 mL), and after adding phthalimide potassium salt (452.28 mg,2.442mmol,2.0 eq), the mixture was stirred at room temperature (20 ℃ C.) for 18hr; TLC (petroleum ether: ethyl acetate=1:1) showed dot formation with increased polarity; the reaction solution was added with water (50 mL), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, washed with water (50 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give 2- [7- (pyridazin-3-yl) heptyl ] isoindole-1, 3-dione (17-2) (185 mg,0.515mmol, 42.17%) as a colourless oil .1H NMR(400MHz,CDCl3)δ9.12(s,1H),8.61(d,J＝3.9Hz,1H),7.84(m,2H),7.71(m,2H),7.20(d,J＝4.7Hz,1H),3.67(m,2H),2.76(m,2H),1.74(dd,J＝15.2,7.8Hz,4H),1.34(m,6H).LC-MS(ESI)[M+H]+＝324.20
The third step, 2- [7- (pyridazin-3-yl) heptyl ] isoindole-1, 3-dione (17-2) (180mg,0.557mmol l,1.0eq) was added to a reaction flask containing ethanol (10 mL), hydrazine hydrate (327.80 mg,5.566mmol,10.0 eq) was added, and the mixture was heated to 50deg.C and stirred for 18hr; the reaction solution was monitored by TLC (dichloromethane: methanol=10:1) for the absence of starting material remaining, with the formation of a dot of increasing polarity; the reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃ C.) and then was substituted with oil pump to give 7- (pyridazin-3-yl) hept-1-amine (17-3) (110 mg,0.512mmol, 92.02%) as a pale yellow oil .1HNMR(400MHz,CDCl3)δ9.05(dd,J＝4.8,1.7Hz,1H),7.39(dd,J＝8.4,4.8Hz,1H),7.32(dd,J＝8.4,1.7Hz,1H),2.98(m,2H),2.68(t,J＝7.0Hz,2H),1.77(m,4H),1.36(m,6H).LC-MS(ESI)[M+H]+＝194.20
The fourth step compound 33- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthrene ] propanoic acid (I) (120 mg,0.267mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (5 mL), HATU (152.51 mg,0.401mmol,1.5 eq) and N, N-diisopropylethylamine (0.133 mL, 0.202 mmol,3.0 eq) were added and stirred at room temperature (20deg.C) for 0.5hr, and then 7- (pyridazin-3-yl) hept-1-amine (17-3) (62.02 mg,0.321mmol,1.2 eq) was added and stirring was continued for 18hr; the reaction liquid is monitored by LC-MS to have no raw material residue; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product 3- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthreyl ] -N- [7- (pyridazin-3-yl) heptyl ] propanamide (17-4) (71 mg,0.102mmol, 38.29%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 624.45
The fifth step compound 3- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthreyl ] -N- [7- (pyridazin-3-yl) heptyl ] propanamide (17-4) (71 mg,0.114mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.138 mL,1.138mmol,10.0 eq) was added and then heated to 50℃and stirred for 18hr; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed that the starting material was essentially reacted; the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridazin-3-yl) heptyl ] propanamide (17) (30 mg,0.052mmol, 46.03%) as a white solid .1H NMR(400MHz,CDCl3)δ9.06(m,1H),7.39(dd,J＝8.4,4.8Hz,1H),7.32(dd,J＝8.4,1.6Hz,1H),5.46(s,1H),3.22(dd,J＝12.8,5.8Hz,3H),2.98(m,2H),2.41(m,1H),2.34(d,J＝5.1Hz,1H),2.22(dd,J＝9.8,5.3Hz,1H),2.14(m,2H),1.92(ddd,J＝15.8,13.4,8.5Hz,3H),1.77(m,3H),1.68(m,3H),1.62(s,3H),1.58(s,2H),1.47(m,2H),1.36(m,5H),1.30(d,J＝10.7Hz,2H),1.25(s,1H),1.12(d,J＝21.3Hz,1H),1.04(m,2H),0.97(d,J＝6.0Hz,2H),0.87(m,2H),0.81(t,J＝7.5Hz,5H).13C NMR(101MHz,CDCl3)δ172.92,163.97,149.55,131.92,128.45,126.40,126.25,79.04,54.29,54.02,53.94,39.46,38.83,37.90,36.94,36.50,36.29,35.61,33.56,32.11,29.60,29.47,29.02,28.96,28.05,27.62,26.73,25.16,21.90,21.79,19.57,15.38,14.29.LC-MS(ESI)[M+H]+＝510.40.
Example 18
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) propanamide
The first step compound 3- (pyridin-2-yl) propan-1-ol (2 g,14.579mmol,1.0 eq) was added to a reaction flask containing dichloromethane (20 mL), and after addition of dess-martin reagent (7.42 g,17.495mmol,1.2 eq) at 0 ℃, the reaction was returned to room temperature (25 ℃) and stirred for 2hr, tlc (petroleum ether: ethyl acetate=1:1) monitored to show the disappearance of starting material; the reaction mixture was quenched by the addition of saturated sodium sulfite solution (20 mL), dichloromethane (20 mL) was added, washed with saturated sodium bicarbonate solution (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried by spin-drying under reduced pressure (water pump, 40 ℃) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-1:1) to give 3- (pyridin-2-yl) propanal (18-1) (1.18 g,7.857mmol, 53.89%) as a colorless oil, LC-MS (ESI) [ m+1] + =136.1
The second step compound 3- (pyridin-2-yl) propanal (18-1) (100 mg,0.74mmol,1.0 eq) and [ (hexahydropyridin-4-ylmethyl) amino ] methanoic acid-2-methylpropan-2-yl ester (190.26 mg,0.88 mmol,1.2 eq) were added to a reaction flask containing tetrahydrofuran (5 mL), and glacial acetic acid (0.008 mL,0.148mmol,0.2 eq) and sodium triacetoxyborohydride (187.28 mg,0.88 mmol,1.2 eq) were added and stirred at room temperature (25 ℃) under nitrogen for 16hr. TLC (dichloromethane: methanol=20:1) monitoring of the reaction showed that a small amount of starting material R2 remained, with the formation of spots of increasing polarity; after adding saturated aqueous sodium bicarbonate (30 mL), dichloromethane (20 mL x 3) was added to extract, the organic phases were combined, dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=1:0 to 9:1) to give the product [ ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) amino ] methanoic acid-2-methylpropan-2-yl ester (18-2) (151 mg, 0.457 mmol, 61.21%) as a pale yellow oil LC-MS (ESI) [ m+1] + =334.2.
The third step compound [ ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) amino ] methanoic acid-2-methylpropan-2-yl ester (18-2) (100 mg, 0.227 mmol,1.0 eq) was added to a reaction flask containing methanol (1 mL), and after adding a methanol solution of hydrogen chloride (0.819 mL,3.274mmol,10eq, 4M), stirred at room temperature (25 ℃ C.) for 16hr; LC-MS monitoring reaction shows that no raw material remains and products are generated; the reaction solution was directly dried under reduced pressure (water pump, 50 ℃ C.) and then was further dried by an oil pump (45 ℃ C., water bath) to give {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methanamine (18-3) (80 mg,0.298mmol, 91.07%) as a yellow oil. LC-MS (ESI) [ m+1] + =234.1.
The fourth step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg, 0.67 mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.074 mL,0.446mmol,2.0 eq) was added and stirring was continued at room temperature (25 ℃) for 0.5hr, followed by {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methanamine (18-3) (54.264 mg, 0.307 mmol,1.2 eq) and stirring was continued; TLC (dichloromethane: methanol=20:1) monitoring the reaction showed the starting material disappeared; the reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) to give the crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) propionamide (18-4) (120 mg,0.151mmol, 67.73%) as a yellow oil. LC-MS (ESI) [ m+1] + = 664.5.
The fifth step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) propanamide (18-4) (45 mg,0.068mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (0.678 mL,0.678mmol,10eq, 1M), the mixture was stirred at 50℃for 16hr; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed a small amount of starting material remaining; the reaction solution was dried under reduced pressure (water pump, 45 ℃) and then added with ethyl acetate (20 mL) and water (20 mL) to dissolve the mixture, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 4) and dried under reduced pressure (water pump, 45 ℃) to give crude product, which was purified by prep-TLC (dichloromethane: methanol=10:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } methyl) propionamide (18) (6.9 mg, 0.0115 mmol, 16.19%) as a white solid .1H NMR(400MHz,CDCl3)δ8.51(d,J＝4.4Hz,1H),7.67(t,J＝7.7Hz,1H),7.21(dd,J＝15.9,10.3Hz,2H),6.16(s,1H),3.73(t,J＝5.9Hz,1H),3.54(m,1H),3.40–3.32(m,3H),3.22(m,3H),3.03(m,2H),2.92(t,J＝7.2Hz,2H),2.65(m,1H),2.54(m,1H),2.44–2.39(m,2H),2.37(m,1H),2.32(m,1H),2.27(m,1H),2.20–2.12(m,2H),1.95(m,3H),1.94–1.88(m,3H),1.87(d,J＝3.9Hz,1H),1.62(s,3H),1.46(m,3H),1.39–1.32(m,1H),1.03(d,J＝7.2Hz,5H),0.97(s,3H),0.88–0.85(m,1H),0.81(dd,J＝13.4,6.6Hz,6H).13C NMR(101MHz,CDCl3)δ173.87,159.06,148.89,137.27,131.82,128.32,123.48,121.93,79.02,77.36,77.25,77.04,76.73,61.93,59.09,54.26,53.90,38.81,37.89,36.92,36.49,35.30,34.58,33.54,32.07,28.07,27.61,25.15,24.16,23.15,21.89,21.78,19.80,19.60,15.41,14.31,13.71.LC-MS(ESI)[M+1]+＝550.4.
Example 19
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {4- [ methyl (methyldioxy-. Lamda.6-thio) amino ] phenyl } butanamide
In the first step, methyl (4-nitrophenyl) amine (1000 mg, 6.578mmol) was dissolved in N, N-dimethylformamide (8 mL), then the mixture was cooled to 0℃under nitrogen, sodium hydride (0.32 g,13.145 mmol) was added to the solution, stirred at 0℃for half an hour, methanesulfonyl chloride (0.90 g,7.886 mmol) was added dropwise, the temperature was raised to room temperature, and stirring was continued for two hours. After the reaction was completed, 30mL of water was added, extracted with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product, and the crude product was purified by a preparation plate (developing agent: petroleum ether: ethyl acetate=5/1) to obtain a pale yellow solid N-methyl-N- (4-nitrophenyl) methanesulfonamide (19-1)(1020mg,4.342mmol,66.06％).1H NMR(400MHz,CDCl3)δ7.82(dd,J＝9.0,2.6Hz,1H),7.66(d,J＝2.6Hz,1H),6.56(d,J＝9.0Hz,1H),4.27(dd,J＝5.7,3.4Hz,2H),3.48–3.44(m,2H),3.05(s,3H).
In the second step, N-methyl-N- (4-nitrophenyl) methanesulfonamide (19-1) (200 mg,0.869 mmol) was dissolved in methanol (10 mL), ammonium formate (273.89 mg, 4.343mmol) and palladium on carbon (50 mg) were added, and stirred at room temperature overnight. After the reaction was completed, the reaction solution was filtered and washed with methanol. Spin-drying the filtrate to obtain crude product, and purifying the crude product with a preparation plate (developing agent: petroleum ether: ethyl acetate=2/1) to obtain yellow solid N- (4-aminophenyl) -N-methyl methane sulfonamide (19-2)(180mg,0.854mmol,98.30％).1H NMR(400MHz,CDCl3)δ7.14(m,2H),6.66(m,2H),3.75(s,2H),3.26(s,3H),2.83(s,3H).
In a third step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylphenanthryl ] butanoic acid (19-2) (50 mg,0.108mmol,1.0 eq), N- (4-aminophenyl) -N-methylmethanesulfonamide (25.96 mg,0.130mmol,1.2 eq), HATU (61.62 mg,0.162mmol,1.5 eq) was dissolved in N, N-dimethylformamide (3 mL), then N, N-diisopropylethylamine (0.054 mL,0.324mmol,3.0 eq) was added thereto, stirred overnight at room temperature, LC-MS was detected, after completion of the reaction was quenched with water, then ethyl acetate (50 mL), the crude product was obtained by washing with saturated aqueous sodium sulfate, drying, washing the dried phase was separated, dried to obtain a crude product, and filtered; ethyl acetate=1:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- {4- [ methyl (methyldioxy- λ6-thio) amino ] phenyl } butanamide (19-3) (68 mg,0.105mmol, yield: 97.57%) as a solid compound .1H NMR(400MHz,CDCl3)δ7.55(d,J＝8.3Hz,2H),7.33(d,J＝8.6Hz,2H),7.13(s,1H),3.30(s,3H),3.18(dd,J＝11.0,4.4Hz,1H),2.83(s,3H),2.34(dd,J＝15.1,7.7Hz,2H),2.13(s,3H),2.03–1.77(m,4H),1.67(d,J＝5.4Hz,5H),1.61(s,3H),1.36(dd,J＝31.6,15.9Hz,4H),1.09–0.97(m,2H),0.89(s,9H),0.87(s,3H),0.85(s,1H),0.80(s,3H),0.75(s,3H),0.03(d,J＝2.4Hz,6H).
In a fourth step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {4- [ methyl (methyldioxy-. Lamda.6-thio) amino ] phenyl } butanamide (19-3) (58 mg,0.090 mmol) was dissolved in tetrahydrofuran (5 mL), and tetrabutylammonium fluoride (235.11 mg,0.899mmol,10.0 eq) was added thereto and stirred at 50℃overnight. LC-MS detection reaction was complete. Quenched with water, then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and spin-dried to give the crude product, which was then purified by preparative plate separation (petroleum ether: ethyl acetate=1:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {4- [ methyl (methyldioxy- λ6-thio) amino ] phenyl } butanamide (19) (16.2 mg,0.028mmol, yield: 31.64%) as a white solid compound .1H NMR(400MHz,CDCl3)δ7.55(d,J＝8.7Hz,2H),7.32(d,J＝8.8Hz,2H),3.30(s,3H),3.22(dd,J＝11.5,4.6Hz,1H),2.84(s,3H),2.35(dd,J＝14.9,7.8Hz,2H),2.15(dd,J＝20.9,5.7Hz,3H),1.94(d,J＝16.2Hz,2H),1.87–1.73(m,3H),1.71–1.64(m,5H),1.61(s,3H),1.44–1.32(m,2H),1.10–1.02(m,2H),0.97(d,J＝6.4Hz,3H),0.90–0.84(m,2H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ171.42,137.28,137.02,132.31,127.86,127.03,120.41,79.10,54.28,53.93,38.82,38.22,37.82,37.47,36.95,36.52,35.11,33.57,32.11,29.70,28.44,28.06,27.64,24.23,21.93,21.85,19.66,15.38,14.30.LC-MS(ESI)[M+H]+＝531.3.
Example 20
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methyl-2, 3-dihydro-1H-indol-5-yl) butanamide
Reference example 2 conversion of 1-methylpyrrolo [2,3-b ] pyridin-5-amine to 1-methyl-2, 3-dihydro-1H-indol-5-amine hydrochloride gives the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methyl-2, 3-dihydro-1H-indol-5-yl) butanamide (20).1H NMR(400MHz,DMSO)δ9.49(s,1H),7.31(s,1H),7.17–7.13(m,1H),6.42(d,J＝8.4Hz,1H),4.32(d,J＝5.1Hz,1H),3.17(t,J＝8.1Hz,2H),2.99(d,J＝6.4Hz,1H),2.82(t,J＝8.1Hz,2H),2.64(s,3H),2.23–2.11(m,3H),2.04–1.83(m,5H),1.67–1.47(m,10H),1.29(dd,J＝20.6,17.9Hz,2H),1.04–0.93(m,2H),0.87(s,3H),0.77(d,J＝8.7Hz,5H),0.68(s,3H).13C NMR(101MHz,DMSO)δ170.69,149.92,133.08,130.78,130.46,126.93,119.01,117.27,107.20,77.38,56.42,54.36,53.98,38.92,38.03,37.04,36.89,36.59,36.53,33.52,32.22,28.78,28.68,28.64,27.90,24.71,22.09,21.90,19.82,16.28,14.56.LC-MS(ESI)[M+H]+＝479.4.
Example 21
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3- { [3- (pyridin-2-yl) propyl ] amino } propyl) propanamide
The first step compound [ (3-aminopropyl) amino ] methanoic acid-2-methylpropan-2-yl ester (3 g,17.218mmol,1.0 eq) and 3-bromoprop-1-yne (2.25 g,18.939mmol,1.1 eq) were added to a reaction flask containing acetonitrile (5 mL), potassium carbonate (5.95 g,43.044mmol,2.5 eq) was added and stirred at room temperature (20deg.C) for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitoring the reaction showed the formation of new spots with small organic phase changes; the reaction mixture was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried under reduced pressure (water pump, 40 ℃) to give the product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 0:1) to give the product { [3- (prop-2-ynylamino) propyl ] amino } methanoic acid-2-methylpropan-2-yl ester (21-1) (700 mg,2.968mmol, 17.24%) as a colorless oil. LC-MS (ESI) [ m+1] + =213.2.
The second step compound { [3- (prop-2-ynylamino) propyl ] amino } methanoic acid-2-methylpropan-2-yl ester (21-1) (560 mg, 2.428 mmol,1.0 eq) and 2-bromopyridine (458.47 mg,2.902mmol,1.1 eq) were added to a reaction flask containing DMF (N, N-dimethylformamide) (1 mL), ditolylphosphoropalladium dichloride (41.05 mg,0.053mmol,1.0eq,0.02 eq), cuprous iodide (5.02 mg,0.026mmol,0.01 eq) and triethylamine (1.83 mL,13.190mmol,5.0 eq) were added and stirred under nitrogen for 16hr at 100 ℃. TLC (dichloromethane: methanol=20:1) monitoring the reaction showed the starting material disappeared; the reaction mixture was taken up in water (30 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with saturated aqueous saline (30 mL x 3) and dried by spin drying under reduced pressure (water pump, 40 ℃) to give crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give [ (3- { [3- (pyridin-2-yl) prop-2-ynyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (21-2) (330 mg,1.026mmol, 38.91%) as a brown viscous substance .1H NMR(400MHz,CDCl3)δ8.53(d,J＝3.7Hz,1H),7.61(td,J＝7.8,1.6Hz,1H),7.40(d,J＝7.8Hz,1H),7.22(d,J＝7.5Hz,1H),5.09(s,1H),3.89(s,2H),3.24(d,J＝5.7Hz,2H),3.03(m,2H),1.92(m,2H),1.33(s,9H).
The third step compound [ (3- { [3- (pyridin-2-yl) prop-2-ynyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (21-2) (300 mg,1.037mmol,1.0 eq) was added to a reaction flask containing methanol (3 mL), palladium on carbon (60 mg,0.056 mmol) was added and stirred at room temperature (20 ℃ C.) under hydrogen atmosphere (15 psi) for 16hr, and no starting material remained as monitored by LC-MS to yield the title product; the reaction solution was filtered through celite, the filter cake was washed with methanol (20 mL), the filtrates were combined and dried under reduced pressure (water pump, 45 ℃) to give the product [ (3- { [3- (pyridin-2-yl) propyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (21-3) (260 mg,0.798mmol, 76.93%) as a brown oil LC-MS (ESI) [ m+1] + =294.3.
The fourth step compound [ (3- { [3- (pyridin-2-yl) propyl ] amino } propyl) amino ] methanoic acid-2-methylpropan-2-yl ester (21-3) (200 mg,0.682mmol,1.0 eq) was added to a reaction flask containing acetonitrile (1 mL) and water (0.5 mL), followed by addition of 9-fluorenylmethyl chloroformate (211.61 mg,0.818mmol,1.2 eq) and sodium carbonate (144.49 mg,1.363mmol,2.0 eq) followed by stirring at room temperature (25 ℃) for 16hr; the reaction was monitored by TLC (petroleum ether: ethyl acetate=3:1) and showed no starting material remained. The reaction mixture was taken up in water (20 mL), extracted with dichloromethane (20 mL. Times.3), the organic phases combined, dried under reduced pressure (water pump, 45 ℃ C.) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:2) to give the product [2, 2-dimethyl-4-oxo-12- (pyridin-2-yl) -5, 9-diaza-3-oxadodecyl-9-yl ] methanoic acid-9H-fluoren-9-ylmethyl ester (21-4) (190 mg, 0.365 mmol, 54.06%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.54(d,J＝4.1Hz,1H),7.75(s,3H),7.56(d,J＝7.3Hz,2H),7.31(dd,J＝22.0,15.5Hz,5H),7.15(d,J＝44.1Hz,1H),4.56(s,2H),4.21(d,J＝5.2Hz,1H),3.26(s,2H),3.09–2.55(m,6H),1.99(s,1H),1.84–1.49(m,4H),1.43(s,9H),1.32(dd,J＝23.9,14.0Hz,2H).
The fifth step compound [2, 2-dimethyl-4-oxo-12- (pyridin-2-yl) -5, 9-diaza-3-oxododecan-9-yl ] methanoic acid-9H-fluoren-9-ylmethyl ester (21-4) (120 mg,0.233mmol,1.0 eq) was added to a reaction flask containing methanol (0.5 mL), followed by addition of a solution of hydrogen chloride in methanol (0.492 mL,2.327mmol,10eq, 4M) and stirring at room temperature (25 ℃ C.) for 16hr; the reaction liquid is monitored by LC-MS, no raw material remains, and products are generated; the reaction solution was dried under reduced pressure (water pump, 45 ℃ C.) and then was further pulled up with an oil pump (water bath, 45 ℃ C.) to give [ (3-aminopropyl) [3- (pyridin-2-yl) propyl ] amino ] methanoic acid-9H-fluoren-9-ylmethyl ester (21-5) (100 mg,0.21 mmol, 93.07%) as a yellow oil. LC-MS (ESI) [ m+1] + =416.2.
The sixth step 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (50 mg,0.111mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (50.84 mg,0.134mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.037 mL,0.223mmol,2.0 eq) were added and stirring was continued at room temperature (25 ℃) for 0.5hr, followed by [ (3-aminopropyl) [3- (pyridin-2-yl) propyl ] amino ] methanoic acid-9H-fluoren-9-ylmethyl ester (21-565 mg, 1.134 mmol,1.2 eq) and stirring was continued at room temperature (25.134 mg, 1.2 hr); the reaction was monitored by TLC (dichloromethane: methanol=20:1) to show no starting material; the reaction mixture was extracted with ethyl acetate (20 mL), and the organic phases were combined and washed with water (20 mL. Times.3). The crude product was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give {11- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -9-oxa-n-1- (pyridin-2-yl) -4, 8-diazaundec-4-yl } methano-9H-fluoren-9-ylmethyl ester (21-6) (90 mg,0.096mmol, 85.91%) as a yellow oil. LC-MS (ESI) [ m+1] + = 846.5 (ms+1).
The seventh step {11- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -9-oxa-no-1- (pyridin-2-yl) -4, 8-diazaundec-4-yl } methanoic acid-9H-fluoren-9-ylmethyl ester (21-6) (90 mg,0.106mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.060 mL,1.060mmol,10eq, 1M) was added and stirred at 50℃for 16hr. LC-MS monitored reaction, TLC (dichloromethane: methanol=10:1) showed no starting material remaining; the reaction solution was dried (water pump, 45 ℃) and dichloromethane (20 mL) was added, washed with water (10 mL x 2), the organic phase was dried under reduced pressure, purified by prep-TLC (dichloromethane: methanol=10:1) and rf=0.23 was collected to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (3- { [3- (pyridin-2-yl) propyl ] amino } propyl) propanamide (21) (8.33 mg,0.015mmol, 14.15%) as a white solid .1H NMR(400MHz,CDCl3)δ8.48(d,J＝3.9Hz,1H),7.74(dd,J＝10.8,4.6Hz,1H),7.34(dd,J＝17.1,7.0Hz,2H),3.46(dd,J＝11.7,6.0Hz,2H),3.27–3.17(m,1H),3.10(t,J＝6.3Hz,4H),3.03(d,J＝6.0Hz,2H),2.39(m,1H),2.30(m,3H),2.22–2.16(m,1H),2.15–2.08(m,3H),1.94(m,2H),1.88(m,3H),1.86(m,2H),1.76–1.68(m,2H),1.65(m,1H),1.62(m,2H),1.59(s,3H),1.34(m,1H),1.12–1.00(m,3H),1.00–0.92(s,3H),0.85(m,2H),0.79(d,J＝8.8Hz,6H).13C NMR(101MHz,CDCl3)δ174.67,159.17,147.75,138.50,131.78,128.26,124.25,122.40,79.09,77.34,77.23,77.02,76.71,54.28,53.94,48.43,45.25,38.83,37.93,36.94,36.51,36.15,35.43,35.35,33.53,32.05,28.07,27.64,26.57,25.19,24.60,21.89,21.78,19.58,15.40,14.33.LC-MS(ESI)[M+1]+＝510.3.
Example 22
Preparation of the Compound N- [7- (1, 2-diazacyclohexen-3-yl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] propanamide
The first step compound, 4-methylpyrimidine (500.00 mg,5.312mmol,1.0 eq), was added to a reaction flask containing tetrahydrofuran (20 mL), and after-78℃lithium diisopropylamide (3.187mL,6.74mmol 1.2eq) was added, the mixture was stirred at-78℃for 30min; 1, 6-dibromohexane (1296.06 mg,5.312mmol,1.0 eq) was added and stirred at room temperature for 18h, and the reaction was monitored by TLC (petroleum ether: ethyl acetate=1:1); the reaction mixture was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product 4- (7-bromoheptyl) pyrimidine (22-1) (450 mg,1.575mmol, 29.64%) as a colorless oily liquid. LC-MS (ESI) [ m+h ] + = 257.10
The second step compound 4- (7-bromoheptyl) pyrimidine (22-1) (450 mg,1.750mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (10 mL), and after adding phthalimide potassium salt (648.18 mg,3.499mmol,2.0 eq), the mixture was stirred at room temperature (20 ℃ C.) for 18hr; TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; the reaction solution was added with water (50 mL), extracted with ethyl acetate (100 mL x 3), the organic phases were combined, washed with water (50 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give 2- [7- (pyrimidin-4-yl) heptyl ] isoindole-1, 3-dione (22-2) (400 mg,1.113mmol, 63.62%) as a colorless oil .1H NMR(400MHz,CDCl3)δ9.12(s,1H),8.61(d,J＝3.9Hz,1H),7.84(m,2H),7.71(m,2H),7.20(d,J＝4.7Hz,1H),3.67(m,2H),2.76(m,2H),1.74(dd,J＝15.2,7.8Hz,4H),1.34(m,6H).LC-MS(ESI)[M+H]+＝324.20
The third step compound 2- [7- (pyrimidin-4-yl) heptyl ] isoindole-1, 3-dione (22-2) (200 mg,0.618mmol,1.0 eq) was added to a reaction flask containing ethanol (10 mL), hydrazine hydrate (364.22 mg,6.184mmol,10.0 eq) was added, and the mixture was heated to 50deg.C and stirred for 18hr; the reaction was monitored by TLC (dichloromethane: methanol=10:1) for no starting material remaining; the reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃ C.) and then was substituted with oil pump to give 7- (pyrimidin-4-yl) hept-1-amine (22-3) (120 mg, 0.560 mmol, 95.37%) as a pale yellow oil .1H NMR(400MHz,CDCl3)δ9.11(s,1H),8.60(d,J＝5.2Hz,1H),7.17(dd,J＝5.2,1.1Hz,1H),2.77(m,4H),1.74(dd,J＝11.0,6.5Hz,2H),1.55(dd,J＝13.7,6.7Hz,2H),1.32(m,6H).LC-MS(ESI)[M+H]+＝194.20
The fourth step compound 33- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthrene ] propanoic acid (22-3) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (5 mL), HATU (127.09 mg, 0.336 mmol,1.5 eq) and N, N-diisopropylethylamine (0.110 mL,0.669mmol,3.0 eq) were added and stirred at room temperature (20deg.C) for 0.5hr, followed by the addition of 7- (pyrimidin-4-yl) hept-1-amine (51.69 mg,0.267mmol,1.2 eq) and stirring continued for 18hr; the reaction liquid is monitored by LC-MS to have no raw material residue; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product 3- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthreyl ] -N- [7- (pyrimidin-4-yl) heptyl ] propanamide (22-4) (74.00 mg,0.107mmol, 47.89%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 492.35
The fifth step compound 3- [ (1R, 2R,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyldeca-hydrophenanthreyl ] -N- [7- (pyrimidin-4-yl) heptyl ] propanamide (22-4) (73 mg,0.117mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.170 mL,1.170mmol,10.0 eq) was added and then heated to 50℃and stirred for 18hr; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed that the starting material was essentially reacted; the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyrimidin-4-yl) heptyl ] propionamide (22) (41 mg,0.069mmol, 59.13%) as a white solid .1H NMR(400MHz,CDCl3)δ8.52(m,1H),7.58(m,1H),7.11(m,2H),3.33(m,1H),3.22(m,2H),2.92(q,J＝9.6Hz,3H),2.78(m,2H),2.31(m,1H),2.17(m,1H),2.06(m,1H),1.84(dd,J＝28.7,25.5Hz,2H),1.69(m,5H),1.62(d,J＝3.6Hz,2H),1.52(m,2H),1.42(d,J＝3.3Hz,1H),1.36(dd,J＝9.4,5.7Hz,5H),1.27(d,J＝12.2Hz,6H),1.17(m,2H),1.03(m,2H),0.96(d,J＝3.5Hz,3H),0.87(d,J＝6.6Hz,2H),0.83(s,2H),0.80(d,J＝3.5Hz,2H),0.78(d,J＝3.4Hz,4H),0.66(m,1H).13C NMR(101MHz,CDCl3)δ173.41,162.43,149.21,149.16,136.31,136.27,122.72,120.96,120.88,79.10,56.18,54.15,54.04,49.22,38.84,38.37,38.33,37.35,36.74,35.37,34.74,33.37,32.20,31.00,29.84,29.78,29.70,29.27,29.05,28.56,28.20,28.13,27.63,27.29,26.80,26.71,25.79,21.42,18.58,15.55,15.49,14.33,12.32.LC-MS(ESI)[M+H]+＝510.40.
Example 23
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (pyridin-2-yloxy) hexyl ] propionamide
In the first step, the compound 1, 6-dibromohexane (2.6 g,10.66mmol,1.0 eq), 2-hydroxypyridine (1.0 g,10.52mmol,1.0 eq), potassium carbonate (1.77 g,12.80mmol,1.2 eq) and potassium iodide (530 mg,3.2mmol,0.3 eq) were dissolved in N-methylpyrrolidone (30 mL), nitrogen was replaced, and the reaction was allowed to proceed at 90 degrees overnight, after which the LC-MS monitoring reaction was completed. Water (100 mL) was added, followed by extraction with ethyl acetate (100 mL), and the organic phase was washed twice with saturated brine (100 mL. Times.2), then dried over anhydrous sodium sulfate, and spin-dried to give the crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 4:1) to give 2- [ (6-bromohexyl) oxy ] pyridine (23-1) (308 mg,1.19mmol, yield as a white solid 11％).1H NMR(400MHz,CDCl3)δ8.23–8.12(m,1H),7.56(m,1H),6.95–6.84(m,1H),6.73(d,J＝8.4Hz,1H),4.29(t,J＝6.6Hz,2H),3.42(t,J＝6.8Hz,2H),1.89(dd,J＝14.0,6.9Hz,2H),1.80(dd,J＝14.0,6.9Hz,2H),1.50(m,4H).LC-MS(ESI)[M+H]+＝258.0.
In the second step, 2- [ (6-bromohexyl) oxy ] pyridine (23-1) (150 mg,0.581mmol,1.0 eq) was dissolved in N, N-dimethylformamide (3 mL), then potassium phthalimide salt (107.61 mg,0.581mmol,1.0 eq) was added thereto, stirred at room temperature overnight, LC-MS was carried out to detect that the reaction was completed, water was added to the reaction solution to kill it, then extracted with ethyl acetate, the organic phase was washed twice with saturated brine and then dried over anhydrous sodium sulfate, filtered, and dried to give a crude product, which was isolated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give 2- [6- (pyridin-2-yloxy) hexyl ] isoindole-1, 3-dione (23-2) (188 mg,0.58mmol, yield: 100%) as an oily compound. LC-MS (ESI) [ m+h ] + =325.1
In the third step, 2- [6- (pyridin-2-yloxy) hexyl ] isoindole-1, 3-dione (23-2) (120 mg,0.370mmol,1.0 eq) was dissolved in ethanol (5 mL), then hydrazine hydrate (0.212 mL,3.699mmol,10 eq) was added thereto, the resulting solution was stirred at 50℃for 3h, LC-MS detection, the solid was filtered off after completion of the reaction, and the filtrate was then dried by spin-drying to give 6- (pyridin-2-yloxy) hex-1-amine (23-3) (60 mg,0.31mmol, yield: 83.48%) as an oily compound. LC-MS (ESI) [ m+h ] + = 195.2.
In a fourth step 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (100 mg,0.223mmol,1.0 eq), 2- (5-aza-2-oxadodecane-12-yl) pyridine (66.96 mg,0.267mmol,1.2 eq), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (127.09 mg, 0.332 mmol,1.5 eq) was dissolved in N, N-dimethylformamide (3 mL), to which N, N-diisopropylethylamine (0.11 mL,0.669mmol,3.0 eq) was then added, stirred overnight at room temperature, and after completion of the reaction, water (10 mL) was added, followed by extraction with ethyl acetate (100 mL), the organic phase was washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and spun-dried to give a crude product, which was then separated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (2-methoxyethyl) -N- [7- (pyridin-2-yl) heptyl ] propanamide (23-4) (82 mg,0.12mmol, 54.03%) as an oily compound .1H NMR(400MHz,CDCl3)δ8.18(d,J＝3.8Hz,1H),7.64(d,J＝7.2Hz,1H),6.92(t,J＝5.9Hz,1H),6.79(d,J＝8.4Hz,1H),5.51(s,1H),4.33(t,J＝6.6Hz,2H),3.21(ddd,J＝15.8,12.0,5.3Hz,3H),2.48–2.38(m,1H),2.34(d,J＝9.1Hz,1H),2.24(m,1H),2.14(dd,J＝8.6,5.7Hz,2H),2.03–1.87(m,3H),1.79(dd,J＝14.3,6.8Hz,3H),1.73–1.65(m,3H),1.62(s,3H),1.60–1.45(m,8H),1.40(dd,J＝14.5,7.9Hz,3H),1.06–0.98(m,2H),0.94–0.85(m,13H),0.78(dd,J＝23.4,7.6Hz,6H),0.07–0.00(m,6H).LC-MS(ESI)[M+H-TBS]+＝493.3.
In a fifth step, the compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (pyridin-2-yloxy) hexyl ] propionamide (23-4) (96 mg,0.154mmol,1.0 eq) was dissolved in tetrahydrofuran (4 mL), then tetrabutylammonium fluoride (402.65 mg,1.54mmol,10 eq) was added thereto in place of nitrogen, 50℃was stirred overnight, and LC-MS monitored for completion of the reaction. Quenched with water, then extracted with ethyl acetate (100 mL), the organic phase was washed twice with saturated brine (50 mL x 2), then dried over anhydrous sodium sulfate, spin dried, and finally isolated and purified (10% methanol/dichloromethane) using preparative plates to give 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (pyridin-2-yloxy) hexyl ] propanamide (23) (19.71 mg,0.038mmol, yield 24.44%) as an oily compound .1H NMR(400MHz,CDCl3)δ8.14(d,J＝3.9Hz,1H),7.62–7.48(m,1H),6.88–6.81(m,1H),6.72(d,J＝8.3Hz,1H),5.50(s,1H),4.27(t,J＝6.6Hz,2H),3.23(m,3H),2.47–2.39(m,1H),2.34(s,1H),2.27–2.19(m,1H),2.14(m,2H),2.03–1.86(m,4H),1.82–1.73(m,4H),1.72–1.66(m,3H),1.62(s,3H),1.54–1.45(m,4H),1.43–1.31(m,3H),1.09–1.01(m,2H),0.99–0.93(m,3H),0.90–0.84(m,2H),0.80(d,J＝8.8Hz,6H).13C NMR(101MHz,CDCl3)δ172.93,163.99,146.88,138.53,131.89,128.49,116.54,111.04,79.05,77.35,77.04,76.72,65.73,54.29,53.94,39.47,38.82,37.89,36.93,36.50,35.59,33.56,32.11,29.62,28.94,28.05,27.62,26.70,25.81,25.14,21.90,21.79,19.58,15.37,14.28.LC-MS(ESI)[M+H]+＝511.4.
Example 24
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylindol-5-yl) amino ] methanamide
Reference example 103, 1-methylbenzo [ d ] [1,2,3] triazapentan-5-amine was changed to 1-methylindole-5-amine to obtain N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylindole-5-yl) amino ] methanamide as a compound (24).1H NMR(400MHz,CDCl3)δ7.44(d,J＝2.0Hz,1H),7.28(m,1H),7.10(d,J＝3.1Hz,1H),7.03(dd,J＝8.6,2.0Hz,1H),6.46(m,1H),6.17(s,1H),4.62(m,1H),3.81(s,3H),3.32(dd,J＝8.6,3.8Hz,1H),3.20(dd,J＝11.4,4.5Hz,1H),3.07(m,1H),2.27(dd,J＝15.7,7.4Hz,1H),2.08(m,2H),1.82(d,J＝11.6Hz,1H),1.73(m,1H),1.66(m,4H),1.60(s,1H),1.53(m,1H),1.43(s,3H),1.32(dd,J＝13.0,3.5Hz,2H),1.27(s,2H),1.00(dd,J＝12.9,3.9Hz,1H),0.96(d,J＝7.9Hz,3H),0.81(m,2H),0.77(s,3H),0.74(d,J＝5.0Hz,3H).13C NMR(101MHz,CDCl3)δ157.74,135.25,130.16,129.85,129.77,129.23,129.00,120.47,118.44,110.04,101.12,79.07,54.17,53.73,38.80,38.37,37.33,36.92,36.42,33.43,33.05,32.06,28.47,28.02,27.59,21.79,21.63,19.57,15.35,14.22.LC-MS(ESI)[M+H]+＝478.30.
Example 25
Preparation of the Compound N- (1, 2-dimethylindol-5-yl) -4- [ (4 aR,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b,8, 10 a-pentamethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] butanamide
In a first step, the reaction mixture, 2-methyl-5-nitro-1H-indole (200 mg,1.135 mmol) and solvent N, N-dimethylformamide (3 mL), were placed in a 50mL three-necked flask, the reaction mixture was placed in an ice bath, the system was filled with nitrogen, sodium hydrogen (54.49 mg,2.270 mmol) was added at 0℃and stirred for 30 minutes, and then methyl iodide (322.26 mg,2.270 mmol) was added at 0℃and the reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was completed, 20mL of water was added thereto, and the mixture was extracted with ethyl acetate (3X 15 mL) and concentrated in vacuo to give a crude product. The crude product was purified by column chromatography over silica gel (petroleum ether: ethyl acetate=3/1) to give 1, 2-dimethyl-5-nitroindole (25-1) as a yellow solid (182 mg,0.957mmol, 84.29%). LC-MS (ESI) [ m+h ] + = 191.1
In a second step, the reaction 1, 2-dimethyl-5-nitroindole (143-1) (140 mg, 0.730 mmol), iron (205.51 mg,3.680 mmol), ammonium chloride (393.72 mg,7.361 mmol) and solvent ethanol (6 mL), water (2 mL) were added to a 50mL round bottom flask and the reaction mixture was stirred at 80deg.C for 2 hours. LC-MS monitoring. After the reaction was completed, the mixture was filtered through celite, and rinsed 3 times with 3mL of ethanol. Concentration in vacuo afforded a brown solid. To the solid was added 30mL of water, extracted with dichloromethane/methanol=10/1 (3×25 mL), dried over anhydrous sodium sulfate and concentrated in vacuo to give crude product which was purified by prep. plate (dichloromethane/methanol=15/1) to give 1, 2-dimethylindol-5-amine (25-2) (90 mg,0.562mmol, 76.31%). LC-MS (ESI) [ m+h ] + =161.1
In a third step, the reactant 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108 mmol), 1, 2-dimethylindol-5-amine (25-2) (20.77 mg,0.130 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (61.62 mg,0.162 mmol), N, N-diisopropylethylamine (0.054 mL,0.324 mmol) was added to solvent N, N-dimethylformamide (3 mL), and the reaction mixture was stirred overnight under nitrogen atmosphere at room temperature under monitoring LC-MS. After completion of the reaction, water (15 mL) and ethyl acetate (3X 10 mL) were added thereto and extracted. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=12/1) to give N- (1, 2-dimethylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (25-3) as a pale yellow solid (50 mg,0.083mmol, 76.49%). LC-MS (ESI) [ m+h ] + = 605.5
In a fourth step, the reaction N- (1, 2-dimethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (25-3) (50 mg,0.083 mmol), tetrabutylammonium fluoride (0.826 mL,0.826 mmol) and solvent tetrahydrofuran (3 mL) were added to a 25mL round bottom flask, and the reaction was stirred at 50℃for 28 hours and monitored by LC-MS. After completion of the reaction, ethyl acetate (15 mL) was added thereto for dilution, and the mixture was washed with water (3X 20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by preparative plate (developer: dichloromethane/methanol=15/1) to give N- (1, 2-dimethylindol-5-yl) -4- [ (4 ar,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b,8, 10 a-pentamethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide as a white solid (25)(30.83mg,0.057mmol,68.63％).1H NMR(400MHz,DMSO)δ9.59(s,1H),7.73(d,J＝1.5Hz,1H),7.25(d,J＝8.7Hz,1H),7.15(dd,J＝8.7,1.8Hz,1H),6.13(s,1H),4.31(d,J＝5.1Hz,1H),3.61(s,3H),3.00(dd,J＝13.7,7.2Hz,1H),2.37(s,3H),2.25(t,J＝7.2Hz,2H),2.16(d,J＝9.9Hz,1H),2.10–2.00(m,2H),1.89(dd,J＝29.2,12.4Hz,3H),1.68–1.48(m,10H),1.38–1.21(m,2H),1.05–0.93(m,2H),0.87(s,3H),0.77(d,J＝7.9Hz,5H),0.68(s,3H).13C NMR(101MHz,DMSO)δ170.89,137.93,134.21,133.11,131.90,127.63,126.93,114.04,110.37,109.21,99.50,77.38,54.38,53.99,38.93,38.04,37.05,36.73,36.54,33.53,32.24,29.70,28.74,28.64,27.90,24.74,22.09,21.90,19.83,16.27,14.57,12.89.LC-MS(ESI)[M+H]+＝491.3.
Example 26
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (pyridin-2-ylamino) hexyl ] propanamide
The first step compound [ (6-aminohexyl) amino ] methanoic acid-2-methylpropan-2-yl ester (800 mg,3.698mmol,1.0 eq) was added to a reaction flask containing DMF (100 mL), and after substitution of nitrogen, cuprous iodide (35.22 mg,0.185mmol,0.05 eq), 2-iodopyridine (758.1 mg,3.698mmol,1 eq), cesium carbonate (2409 mg, 7.390 mmol,2 eq), 2- (2-methylpropanoyl) cyclohex-1-one (124.4 mg,0.740mmol,0.2 eq) was added and stirred at 40℃for 21hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; ethyl acetate (200 mL) was added to the reaction solution, which was washed with water (200 mL x 2), dried under reduced pressure (pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1), and the product { [6- (pyridin-2-ylamino) hexyl ] amino } methanoic acid-2-methylpropan-2-yl ester (26-1) (300 mg,0.920mmol, 24.88%) was collected as a colorless oily liquid. LC-MS (ESI) [ m+h ] + = 294.41.
The second step compound { [6- (pyridin-2-ylamino) hexyl ] amino } methanoic acid-2-methylpropan-2-yl ester (26-1) (100 mg,0.307mmol,1.0 eq) was added to a reaction flask containing 4mol/L of 1-4-dioxane in hydrochloric acid (5 mL) and stirred for 2hr. LC-MS showed that after completion of the reaction, the reaction was dried under reduced pressure (water pump, 40 ℃ C.) to give crude 6- (pyridin-2-ylamino) hex-1-amine (26-2) as a white solid which was directly added to the next step (60 mg,0.310mmol, 100%). LC-MS (ESI) [ m+h ] + = 194.16.
The third step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (160 mg, 0.356 mmol,1.0 eq)) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg, 0.67 mmol,1.2 eq) and diisopropylethylamine (0.074 mL, 0.4476 mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by 6- (pyridin-2-ylamino) hex-1-amine (26-2) (60 mg,0.310mmol,1 eq); TLC (dichloromethane: methanol=20:1) monitored the reaction; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (pyridin-2-ylamino) hexyl ] propionamide (26-3) (60 mg,0.077mmol, 14.97%) as a pale yellow solid which was directly fed to the next step;
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (pyridin-2-ylamino) hexyl ] propanamide (26-3) (60 mg,0.077mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.445 mL,1.445mmol,10 eq) was added and then warmed to 50℃and stirred for 16hr; TLC (dichloromethane: methanol=20:1) monitoring of the reaction showed that the starting material was essentially reacted. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [6- (pyridin-2-ylamino) hexyl ] propionamide (26) (17.32 mg,0.034mmol, 44.15%) as a white solid .1H NMR(400MHz,CDCl3)δ8.02(d,J＝4.2Hz,1H),7.49–7.43(m,1H),6.59–6.55(m,1H),6.42(d,J＝8.5Hz,1H),5.51(s,1H),5.05(s,1H),3.28–3.20(m,5H),2.47–2.34(m,2H),2.24(s,1H),2.15(dd,J＝11.5,4.9Hz,3H),1.91(d,J＝4.3Hz,4H),1.78–1.64(m,6H),1.62(s,3H),1.53–1.48(m,2H),1.46–1.41(m,2H),1.37(dd,J＝11.5,4.9Hz,3H),1.25(s,1H),1.04(t,J＝8.1Hz,2H),0.97(d,J＝6.1Hz,3H),0.85(dd,J＝7.4,4.9Hz,2H),0.80(d,J＝8.9Hz,6H).13C NMR(101MHz,CDCl3)δ172.95,158.20,146.56,138.30,131.91,128.46,112.49,106.93,79.06,54.29,53.94,42.10,39.29,38.83,37.81,36.93,36.51,35.61,33.57,32.11,29.58,29.25,28.05,27.63,26.57,26.53,25.16,21.91,21.80,19.58,15.37,14.29.LC-MS(ESI)[M+H]+＝510.4.
Example 27
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [ (1-methylindol-5-yl) methyl ] propanamide
The first step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (67.79 mg,0.178mmol,1.0 eq) and N, N-Diisopropylethylamine (DIEA) (0.029 mL,0.178mmol,1.0 eq) were added, followed by stirring at room temperature (25 ℃) for 0.5hr, and stirring was continued after adding (1-methylindol-5-yl) methanamine (28.56 mg,0.178mmol,1 eq) for 1hr; the reaction was monitored by TLC (petroleum ether: ethyl acetate=3:1) for completion. Water (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL. Times.3), washed with saturated aqueous saline (20 mL. Times.3), and the organic phase was dried by spin-drying under reduced pressure (water pump, 40 ℃ C.) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-6:1) to give 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -N- [ (1-methylindol-5-yl) methyl ] propionamide (27-1) (55 mg,0.084mmol, 46.99%) as an off-white solid .1H NMR(400MHz,CDCl3)δ7.52(s,1H),7.29(d,J＝8.5Hz,1H),7.15(d,J＝8.5Hz,1H),7.06(d,J＝3.0Hz,1H),6.45(d,J＝2.9Hz,1H),5.68(s,1H),4.51(d,J＝5.2Hz,2H),3.79(s,3H),3.17(dd,J＝11.0,4.5Hz,1H),2.57–2.23(m,5H),2.20–2.06(m,3H),1.95(d,J＝14.3Hz,3H),1.81(d,J＝14.7Hz,2H),1.68(d,J＝16.2Hz,3H),1.58(s,3H),1.35–1.31(m,1H),1.01–0.94(m,3H),0.89(d,J＝3.9Hz,8H),0.86(s,3H),0.76(d,J＝16.4Hz,6H),0.03(d,J＝2.9Hz,6H).
The second step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (1-methylindol-5-yl) methyl ] propanamide (27-1) (50 mg,0.085mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (0.846 mL,0.846mmol,10eq, 1M) was added thereto, followed by heating to 50℃and stirring for 16hr. The reaction was monitored by TLC (petroleum ether: ethyl acetate=2:1). The reaction solution was dried under reduced pressure, ethyl acetate (10 mL) and water (20 mL) were added and mixed well, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) successively to give a crude product which was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=2:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (1-methylindole-5-yl) methyl ] propionamide (27) (24.49 mg,0.046mmol, 54.89%) as a white solid .1H NMR(400MHz,CDCl3)δ7.52(s,1H),7.29(d,J＝8.4Hz,1H),7.14(d,J＝8.4Hz,1H),7.06(d,J＝2.9Hz,1H),6.45(d,J＝2.5Hz,1H),5.71(s,1H),4.51(d,J＝5.0Hz,2H),3.79(S,3H),3.20(m,1H),2.46(m,1H),2.40–2.25(m,2H),2.23–2.11(m,2H),1.94(s,2H),1.87–1.71(m,3H),1.70–1.59(m,4H),1.58(s,3H),1.40–1.23(m,3H),1.07–0.99(m,2H),0.98–0.92(m,3H),0.87(d,J＝12.6Hz,1H),0.84–0.73(m,6H).13C NMR(101MHz,CDCl3)δ172.52,136.19,131.79,129.51,129.06,128.62,121.95,120.40,109.50,100.86,79.07,77.34,77.23,77.03,76.71,54.27,53.92,44.38,38.82,37.85,36.92,36.49,35.51,33.51,32.93,32.11,28.03,27.62,25.12,21.88,21.70,19.55,15.36,14.27.LC-MS(ESI)[M+1]+＝477.3.
Example 28
Preparation of the Compound N- (3-chloro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
Reference example 2, 1-methylpyrrolo [2,3-b ] pyridin-5-amine was changed to 3-chloro-1-methylindol-5-amine to give N- (3-chloro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (28).1H NMR(400MHz,CDCl3)δ5.82(s,1H),3.46(s,4H),3.36(s,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.23–2.11(m,3H),2.06(dd,J＝17.8,9.5Hz,2H),1.93(m,3H),1.81–1.62(m,6H),1.59(s,3H),1.55(m,1H),1.42–1.26(m,2H),1.10–1.00(m,2H),0.97(s,3H),0.87(m,2H),0.80(d,J＝9.0Hz,6H).13C NMR(101MHz,CDCl3)δ171.23,133.28,132.48,130.93,127.74,126.10,125.78,117.14,110.03,109.81,104.34,79.13,54.31,53.98,38.84,37.85,37.54,36.98,36.54,33.59,33.11,32.14,28.54,28.06,27.66,24.49,21.94,21.88,19.69,15.37,14.31.LC-MS(ESI)[M+H]+＝511.3.
Example 29
Preparation of 6- (pyrazin-2-yl) hexyl ester of the compound ({ 2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) methanoate
The first step compound, 2-bromopyrazine (1000.00 mg,6.290mmol,1.0 eq) and 5-hexyn-1-ol (740.73 mg, 7.268 mmol,1.2 eq) were added to a reaction flask containing acetonitrile (20 mL), and after addition of cuprous iodide (71.87 mg,0.377mmol,0.06 eq), ditriphenylphosphine palladium dichloride (146.82 mg,0.189mmol,0.03 eq) and triethylamine (4.371 mL,31.449mmol,5.0 eq) under nitrogen protection, stirred at room temperature (20 ℃) for 18hr. Filtering, and drying the filtrate under reduced pressure (water pump, 40 ℃) to obtain white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1), and the product 6- (pyrazin-2-yl) hex-5-yn-1-ol (29-1) (920 mg,4.699mmol, 74.70%) was obtained as a colorless oily compound .1H NMR(400MHz,CDCl3)δ8.61(d,J＝1.4Hz,1H),8.50(dd,J＝2.4,1.6Hz,1H),8.44(d,J＝2.5Hz,1H),3.71(m,2H),2.55(m,2H),1.76(dt,J＝6.5,3.2Hz,4H),1.67(s,2H).
The second step compound 6- (pyrazin-2-yl) hex-5-yn-1-ol (29-1) (920 mg,5.221mmol,1.0 eq) was added to a reaction flask containing ethanol (20 mL), pd/C (55.56 mg,0.522mmol,0.1 eq) was added after hydrogen replacement, and stirred at 50℃for 18hr. The celite was filtered and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give the product 6- (pyrazin-2-yl) hex-1-ol (29-2) (600 mg,3 719 mmol, 63-76%) as a colorless oil .1H NMR(400MHz,CDCl3)δ8.49(m,1H),8.46(d,J＝1.3Hz,1H),8.40(d,J＝2.5Hz,1H),3.64(t,J＝6.5Hz,2H),2.82(m,2H),1.77(m,2H),1.58(m,2H),1.42(dd,J＝7.0,3.5Hz,4H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (50 mg,0.111mmol,1.0 eq) was added to a reaction flask containing toluene (2 mL), diphenyl azide phosphate (32.20 mg,0.117mmol,1.05 eq) and triethylamine (0.31 mL,0.223mmol,2.0 eq) were stirred at room temperature (90 ℃) for 1hr, the reaction mixture was cooled to room temperature, and 6- (pyrazin-2-yl) hex-1-ol (29-2) (100.42mg, 0.557mmol, 5.eq) and DBU (47.24mg, 0.557 eq) were further stirred at room temperature (90 ℃) for 18hr; LC-MS monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product ({ 2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) methanoic acid-6- (pyrazin-2-yl) hexyl ester (29-3) (26 mg,0.037mmol, 33.55%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 494.30
The fourth step compound ({ 2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) was added to a reaction flask containing tetrahydrofuran (5 mL), and after adding tetrabutylammonium fluoride (0.479 mL,0.479mmol,10.0 eq) was stirred at 50℃for 18hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product ({ 2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) methanoic acid-6- (pyrazin-2-yl) hexyl ester (29) (4 mg,0.008mmol, 15.86%) as a white solid .1H NMR(400MHz,CDCl3)δ8.54(s,3H),4.66(s,1H),4.04(t,J＝6.3Hz,2H),3.22(dd,J＝11.5,4.6Hz,2H),3.04(d,J＝7.7Hz,1H),2.83(m,2H),2.28(m,2H),2.21(s,4H),1.96(m,4H),1.76(dd,J＝9.4,3.7Hz,2H),1.67(m,3H),1.63(s,3H),1.41(d,J＝7.7Hz,3H),1.35(m,1H),1.28(d,J＝11.2Hz,2H),1.06(m,2H),0.96(s,3H),0.86(m,2H),0.80(t,J＝7.5Hz,6H).13C NMR(101MHz,CDCl3)δ156.76,144.29,141.85,129.73,79.09,64.71,54.24,53.87,39.68,38.82,38.00,36.93,36.50,35.34,33.52,32.16,29.55,29.32,29.26,28.92,28.04,27.61,27.21,25.66,21.88,21.78,19.67,15.37,14.26.LC-MS(ESI)[M+H]+＝512.35.
Example 30
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7, 10-dihydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-2-yl) heptyl ] propionamide
In the first step, 7- (pyridin-2-yl) heptanal (19-2) (150 mg,0.784mmol,1.0 eq) and 2-methoxyethyl-1-amine (58.90 mg,0.784mmol,1.0 eq) were dissolved in tetrahydrofuran (5 mL), then acetic acid (0.009 mL,0.157mmol,0.2 eq) and sodium borohydride acetate (198.50 mg,0.941mmol,1.2 eq) were added thereto, nitrogen was replaced, and then stirred at room temperature overnight. After completion of the reaction, saturated sodium hydrogencarbonate (20 mL) was added thereto to be quenched, followed by extraction with ethyl acetate (100 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by column chromatography (dichloromethane: methanol=1:0 to 10:1) to give 2- (5-aza-2-oxadodecane-12-yl) pyridine (30-1) (100 mg,0.36mmol, 45.83%) as an oily compound. LC-MS (ESI) [ m+h ] + = 251.2.
In a second step 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (30-1) (100 mg,0.223mmol,1.0 eq), 2- (5-aza-2-oxododecane-12-yl) pyridine (66.96 mg,0.267mmol,1.2 eq), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (127.09 mg,0.334mmol,1.5 eq) was dissolved in N, N-dimethylformamide (3 mL), then N, N-diisopropylethylamine (0.11 mL,0.669mmol, 3.eq) was added thereto, and the mixture was stirred at room temperature, and the reaction mixture was washed with water (10 mL, 50mL of saturated aqueous solution, dried phase was washed with water (dry ethyl chloride, 50mL of saturated aqueous phase, dry phase was obtained), then isolated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (2-methoxyethyl) -N- [7- (pyridin-2-yl) heptyl ] propanamide (30-2) (82 mg,0.12mmol, 54.03%) as an oily compound .1H NMR(400MHz,CDCl3)δ8.54(d,J＝4.8Hz,0H),7.68(dd,J＝13.7,7.2Hz,0H),7.20(d,J＝7.6Hz,0H),3.52–3.49(m,0H),3.45(d,J＝2.9Hz,0H),3.34(s,0H),3.33(d,J＝2.6Hz,0H),3.32–3.27(m,0H),3.17(dd,J＝11.2,4.7Hz,0H),2.87–2.82(m,0H),2.37(d,J＝5.7Hz,0H),2.18(dd,J＝17.8,8.6Hz,0H),2.04–1.85(m,1H),1.75(s,0H),1.61(s,0H),1.40–1.33(m,1H),1.31–1.23(m,1H),1.10–0.95(m,0H),0.89(s,9H),0.87(d,J＝3.1Hz,0H),0.80(d,J＝1.9Hz,0H),0.74(d,J＝4.0Hz,0H),0.03(d,J＝3.1Hz,1H).
In a third step, the compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (2-methoxyethyl) -N- [7- (pyridin-2-yl) heptyl ] propionamide (30-2) (72 mg,0.106mmol,1.0 eq) was dissolved in THF (5 mL), then tetrabutylammonium fluoride (276.38 mg,1.057 mmol) was added thereto, nitrogen was replaced, then warmed to 50℃overnight with stirring, LC-MS detection, water was added after completion of the reaction to kill, the organic phase was then extracted with ethyl acetate (100 mL), washed twice with saturated brine (50 mL), washed off tetrabutylammonium fluoride, dried over anhydrous sodium sulfate, filtered, spin-dried, and then separated and purified via prep plate (dichloromethane/methanol=10:1) to give 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-methoxyethyl) -N- [7- (pyridin-2-yl) heptyl ] propionamide (30) (14.4 mg,0.024mmol, 22.89%) as an oily compound .1H NMR(400MHz,CDCl3)δ8.55(d,J＝4.9Hz,1H),7.71(s,1H),7.23(s,2H),3.50(dd,J＝6.1,3.7Hz,2H),3.45(d,J＝2.9Hz,2H),3.34(d,J＝4.0Hz,3H),3.30(d,J＝7.6Hz,2H),3.23(s,1H),2.85(s,2H),2.37(d,J＝5.1Hz,2H),2.21(m,2H),2.00–1.88(m,3H),1.76(m,3H),1.67(s,2H),1.61(s,3H),1.40–1.23(m,13H),1.05(d,J＝9.0Hz,3H),0.96(d,J＝2.8Hz,3H),0.87(d,J＝9.7Hz,1H),0.81(s,3H),0.79–0.75(m,3H).13C NMR(101MHz,CDCl3)δ172.97,162.23,149.18,149.13,136.31,132.29,128.08,122.70,120.96,120.88,79.06,77.35,77.03,76.71,71.12,70.87,59.11,58.80,54.31,53.95,49.33,45.91,38.82,38.36,38.32,36.95,36.51,33.47,32.16,29.84,29.79,29.32,29.28,29.12,28.06,27.71,27.63,26.96,26.81,25.18,21.94,21.76,19.53,15.38,14.29.LC-MS(ESI)[M+H]+＝567.5.
Example 31
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- { [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } propanamide
The first step compound hexahydropyridine amino-2-methylpropan-2-yl ester (500 mg,2.333mmol,1.0 eq) and pyridine-2-carbaldehyde (249.90 mg,2.333mmol,1.0 eq) were added to a reaction flask containing tetrahydrofuran (5 mL), and glacial acetic acid (0.027 mL,0.467mmol,0.2 eq) and sodium triacetoxyborohydride (590.54 mg,2.800mmol,1.2 eq) were added and stirred at room temperature (25 ℃ C.) under nitrogen for 16hr. TLC (dichloromethane: methanol=20:1) monitored reaction was complete; after adding saturated aqueous sodium bicarbonate (30 mL), dichloromethane (20 mL x 3) was added to extract, the organic phases were combined, dried by spin drying under reduced pressure (water pump, 40 ℃) to give crude product which was purified by column chromatography (dichloromethane: methanol=1:0 to 9:1) to give product ({ [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } amino) methanoic acid-2-methylpropan-2-yl ester (31-1) (270 mg,0.884mmol, 37.89%) as a pale yellow oil .1H NMR(400MHz,CDCl3)δ8.56(d,J＝4.1Hz,1H),7.67(td,J＝7.7,1.6Hz,1H),7.49(s,1H),7.23–7.15(m,1H),4.60(s,1H),3.74(s,2H),3.01(dd,J＝15.0,8.9Hz,4H),2.19(dd,J＝21.3,11.0Hz,2H),1.69(d,J＝12.4Hz,2H),1.42(d,J＝6.6Hz,12H).
The second step compound ({ [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } amino) methanoic acid-2-methylpropan-2-yl ester (31-1) (100 mg,0.327mmol,1.0 eq) was added to a reaction flask containing methanol (1 mL), and after addition of a solution of hydrogen chloride in methanol (0.819 mL,3.274mmol,10eq, 4M), stirred at room temperature (25 ℃ C.) for 16hr; LC-MS monitoring reaction showed no raw material remaining; the reaction solution was dried under reduced pressure (water pump, 50 ℃ C.) and then was further dried by an oil pump (45 ℃ C., water bath) to give a yellow oily substance of [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methane amine (31-2) (80 mg,0.298mmol, 91.07%) as a product .1H NMR(400MHz,DMSO)δ10.52(s,1H),8.73–8.64(m,1H),8.14(s,2H),7.95(td,J＝7.7,1.8Hz,1H),7.69(d,J＝7.8Hz,1H),7.56–7.44(m,1H),4.45(s,2H),3.40(d,J＝10.3Hz,2H),3.06(t,J＝12.0Hz,2H),2.74(s,2H),1.93(d,J＝13.1Hz,3H),1.60(d,J＝11.5Hz,2H).
The third step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.074 mL,0.446mmol,2.0 eq) were added and stirring was continued at room temperature (25 ℃) for 0.5hr, followed by addition of [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methanamine (31-2) (54.90 mg,0.26 mmol,1.2 eq) and stirring was continued for 1hr; TLC (dichloromethane: methanol=20:1) monitoring the reaction showed the starting material disappeared; the reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) to give the crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silylhydroxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- { [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } propionamide (31-3) (120 mg,0.151mmol, 67.73%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.57(d,J＝4.1Hz,1H),7.71–7.62(m,1H),7.56–7.45(m,1H),7.23–7.16(m,1H),5.55(s,1H),3.76(s,2H),3.23–3.09(m,3H),3.01(d,J＝10.1Hz,2H),2.49–2.38(m,1H),2.37–2.30(m,1H),2.24(ddd,J＝27.7,13.7,9.0Hz,3H),2.16–2.08(m,2H),2.05–1.92(m,3H),1.91–1.85(m,2H),1.71(d,J＝3.1Hz,2H),1.66(d,J＝11.2Hz,3H),1.63(d,J＝4.9Hz,3H),1.59–1.51(m,3H),1.35–1.23(m,4H),1.08–0.97(m,3H),0.90–0.85(m,11H),0.79(s,3H),0.75(d,J＝6.1Hz,3H),0.03(t,J＝6.1Hz,6H).
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- { [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } propionamide (31-3) (110 mg,0.173mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.729 mL,1.729mmol,10eq, 1M) was added and then warmed to 50℃and stirred for 16hr; TLC (dichloromethane: methanol=10:1) monitored the reaction was essentially complete; the reaction solution was dried under reduced pressure (water pump, 45 ℃) and then added with ethyl acetate (20 mL) and water (20 mL) to dissolve the mixture, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 4) and dried under reduced pressure (water pump, 45 ℃) to give crude product, which was purified by prep-TLC (dichloromethane: methanol=10:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- { [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methyl } propionamide (31.05 mg,0.059mmol, 34.13%) as a white solid .1H NMR(400MHz,CDCl3)δ8.56(d,J＝4.7Hz,1H),7.66(td,J＝7.7,1.6Hz,1H),7.44(d,J＝7.7Hz,1H),7.18(dd,J＝6.8,5.3Hz,1H),5.55(dd,J＝27.1,13.5Hz,1H),3.68(s,2H),3.22(dd,J＝11.5,4.6Hz,1H),3.12(dt,J＝15.1,7.1Hz,2H),2.95(d,J＝11.2Hz,2H),2.55–2.40(m,1H),2.39–2.18(m,3H),2.17–2.07(m,4H),2.05–1.95(m,2H),1.94–1.86(m,2H),1.80–1.72(m,2H),1.67(d,J＝7.7Hz,3H),1.62(s,3H),1.59(m,1H),1.56–1.50(m,1H),1.39(m,3H),1.05(m,2H),1.00–0.94(s,3H),0.92–0.84(m,2H),0.81(dd,J＝9.8,4.7Hz,6H).13C NMR(101MHz,CDCl3)δ173.05,149.22,136.47,131.82,128.55,124.22,123.48,122.17,79.04,77.35,77.23,77.03,76.71,64.60,54.28,53.91,53.49,44.93,38.82,37.83,36.92,36.50,35.72,35.54,33.57,32.11,29.65,28.05,27.62,25.11,21.89,21.76,19.59,15.38,14.28.LC-MS(ESI)[M+1]+＝522.4.
Example 32
Preparation of the Compound 4- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [1- (pyrazin-2-ylmethyl) indol-5-yl ] butanamide
In a first step, under nitrogen, the reaction pyrazin-2-yl-methanol (200 mg,1.816 mmol), triethylamine (0.505 mL,3.632 mmol) and solvent dichloromethane (6 mL) were added to a 50mL three-necked flask, cooled to 0℃and methylsulfonyl chloride (416.01 mg,3.632 mmol) was added and the reaction stirred at 0deg.C for 3hr. LC-MS monitored the reaction. After completion of the reaction, the reaction solution was diluted with methylene chloride. Washed with water and saturated brine. The dichloromethane layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the product pyrazin-2-ylmethyl methanesulfonate (32-1) (150 mg, 0.719 mmol, 39.50%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =189.1
In a second step, the reaction mixture 5-nitro-1H-indole (32-1) (100 mg, 0.611 mmol) and solvent N, N-dimethylformamide (6 mL) were added to a 50mL three-necked flask, the temperature was reduced to 0℃and sodium hydride (29.62 mg,1.234 mmol) was added, followed by stirring for 0.5hr, and then the resulting product pyrazin-2-ylmethyl methanesulfonate ((139.28 mg,0.740 mmol) was added, the reaction mixture was stirred at room temperature under N2 protection for 3hr.LC-MS monitoring, water (15 mL) was added, extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated brine (10 mL. Times.2) and then dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give the product 5-nitro-1- (pyrazin-2-ylmethyl) indole (32-2) (150 mg,0.53 mol, 86.10%) as colorless oil.
In a third step, the reaction 5-nitro-1- (pyrazin-2-ylmethyl) indole (32-2) (100 mg,0.393 mmol), palladium on carbon 10% (20 mg,0.188 mmol) and solvent methanol (8 mL) were added to a 50mL three-necked flask, and the reaction was stirred at room temperature for 2H under H2 protection. LC-MS monitoring, after the reaction is completed, the reaction solution is filtered by suction, and the organic phase is dried by pulling and separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1), so as to obtain the product 1- (pyrazin-2-ylmethyl) indol-5-amine (32-3) (60 mg,0.214mmol, 54.42%) as pale yellow oil. LC-MS (ESI) [ m+h ] + =225.1
In a fourth step, the reaction 44- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (0.054 mL,0.324 mmol), N, N-diisopropylethylenediamine (20.71 mg,0.108 mmol), 1-hydroxybenzotriazole (32-3) (14.60 mg,0.108 mmol) and solvent N, N dimethylformamide (6 mL) were added under nitrogen protection in a 50mL three-necked flask, and the reaction mixture was stirred at room temperature for 8hr under N2 protection. LC-MS monitored reaction was complete. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give the crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [1- (pyrazin-2-ylmethyl) indol-5-yl ] butanamide (32-4) (20 mg,0.027mmol, 24.90%) as a pale yellow oil.
In a fifth step, the reaction 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [1- (pyrazin-2-ylmethyl) indol-5-yl ] butanamide (20 mg,0.030 mmol), tetrabutylammonium fluoride (39.22 mg,0.150 mmol) and the solvent tetrahydrofuran (2 mL) were added in a 50mL three-necked flask, and the reaction mixture was stirred under N 2 at 50℃for 8hr. LC-MS monitored the reaction. After the reaction, the tetrahydrofuran is concentrated and removed, and crude products are separated and purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to obtain products 4- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (pyrazin-2-ylmethyl) indol-5-yl ] butyramide (32)(10mg,0.017mmol,57.30％).1H NMR(400MHz,CDCl3)δ8.54(s,1H),8.48(s,1H),8.13(s,1H),7.85(s,1H),7.24-7.19(m,4H),6.55(d,J＝3.0Hz,1H),5.45(s,2H),3.23-3.20(m,1H),2.45–2.27(m,2H),2.18-2.15(m,3H),1.92-1.82(m,5H),1.77–1.61(m,9H),1.41–1.21(m,2H),1.14–1.01(m,2H),0.96(s,3H),0.88–0.75(m,8H).13C NMR(101MHz,CDCl3)δ171.22,152.78,144.11,143.88,143.05,133.49,132.50,130.80,129.05,128.98,127.69,116.47,113.12,109.55,102.85,79.11,54.30,53.97,50.13,38.83,37.86,37.53,36.97,36.53,33.58,32.13,28.55,28.07,27.65,24.52,21.93,21.87,19.68,15.39,14.30.LC-MS(ESI)[M+H]+＝555.3.
Example 33
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-methyl-N- [7- (pyridin-2-yl) heptyl ] propanamide
The first step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (141.37 mg,0.315mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (143.74 mg,0.378mmol,1.2 eq) and diisopropylethylamine (0.104 mL,0.630mmol,2.0 eq) were added and stirring was continued at room temperature (20 ℃) for 0.5hr, followed by methyl [7- (pyridin-2-yl) heptyl ] amine (19-3) (65 mg,0.315mmol,1.0 eq) and stirring continued for 18hr; TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-methyl-N- [7- (pyridin-2-yl) heptyl ] propionamide (33-1) (83 mg,0.117mmol, 37.22%) as a pale yellow oil; LC-MS (ESI) [ m+h ] + = 505.40
The second step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-methyl-N- [7- (pyridin-2-yl) heptyl ] propanamide (33-1) (83 mg,0.130mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.303 mL,1.303mmol,10.0 eq) was added and the mixture was warmed to 50℃and stirred for 18hr; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed the reaction was essentially complete; the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by prep-TLC (dichloromethane: methanol=10:1), to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N-methyl-N- [7- (pyridin-2-yl) heptyl ] propanamide (33) (11 mg,0.021mmol, 15.99%) as a white solid .1H NMR(400MHz,CDCl3)δ8.52(d,J＝3.9Hz,1H),7.60(t,J＝7.6Hz,1H),7.17–7.01(m,2H),3.37–3.30(m,1H),3.23(m,2H),2.93(m,3H),2.78(t,J＝7.0Hz,2H),2.35(d,J＝9.9Hz,2H),2.21(m,3H),1.96(m,8H),1.72(m,6H),1.61(s,3H),1.55–1.46(m,2H),1.36(d,J＝4.3Hz,4H),1.28(d,J＝10.2Hz,2H),1.06(m,2H),0.96(d,J＝3.5Hz,3H),0.87(s,1H),0.81(d,J＝2.0Hz,3H),0.78(d,J＝3.4Hz,3H).13C NMR(101MHz,CDCl3)δ172.46,148.66,135.98,120.52,78.63,77.35,77.03,76.91,76.72,76.60,76.28,53.87,38.38,37.84,36.52,36.07,33.06,31.70,29.36,28.84,27.63,24.80,21.34,19.08,14.94,13.85.LC-MS(ESI)[M+H]+＝523.40
Example 34
Preparation of the Compound 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [5- (naphthalen-2-yl) pentyl ] propionamide
First step potassium carbonate (2810 mg,20.301mmol,2.0 eq) was added to a reaction flask containing acetonitrile (40 mL), naphthalene-2-carbaldehyde (1590 mg,10.151mmol,1.0 eq) and methyl 4- (bromotriphenyl-. Lamda.5-methylphosphine) butyrate (900 mg,20.301mmol,2.0 eq) were added, followed by stirring at 90℃for 4hr; TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. Ethyl acetate (20 mL) was added to the reaction solution, which was washed with water (20 mL x 2), dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 8:2) and the product (4E) -5- (naphthalen-2-yl) pent-4-enoic acid methyl ester (34-1) (190 mg,6.325mmol, 62.32%) was collected as a white solid .1H NMR(400MHz,CDCl3)δ7.79(ddd,J＝20.6,10.7,4.8Hz,3H),7.69(d,J＝16.0Hz,1H),7.60–7.38(m,3H),6.61(dd,J＝13.1,11.2Hz,1H),5.71(dt,J＝11.6,7.2Hz,1H),3.68(d,J＝13.7Hz,3H),2.79–2.72(m,1H),2.59(t,J＝6.5Hz,1H),2.52–2.45(m,1H).
The second step of the compound (4E) -methyl 5- (naphthalen-2-yl) pent-4-enoate (34-1) (550 mg,2.289mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), lithium aluminum hydride (173.72 mg,4.578mmol,2.0 eq) was added under ice bath, and the mixture was returned to room temperature and stirred for 2hr. TLC (petroleum ether: ethyl acetate=2:1) monitored the reaction; water (15 ml x 3) was added for washing, ethyl acetate (15 ml x 3) was extracted, and dried under reduced pressure to give crude silica gel column chromatography (petroleum ether: ethyl acetate=1:0 to 8:2) for purification to give the product (4E) -5- (naphthalen-2-yl) pent-4-en-1-ol (34-2) (390 mg,1.470mmol, 64.21%) as a white solid LC-MS (ESI) [ m+h ] + =213.
The third step compound (4E) -5- (naphthalen-2-yl) pent-4-en-1-ol (34-2) (420 mg,1.978mmol,1.0 eq) was added to a reaction flask containing methanol (10 mL), palladium on carbon (20 mg) was added, replaced with hydrogen three times, and stirred at room temperature (18 ℃ C.) for 16hr; LC-MS monitors the reaction of the raw materials; the reaction solution was filtered through celite and dried under reduced pressure (water pump, 50 ℃ C.) to give the product 5- (naphthalen-2-yl) pentan-1-ol (34-3) (412 mg, 1.178 mmol, 77.75%) as a white solid; LC-MS (ESI) [ m+h ] + =215.
The fourth step compound 5- (naphthalen-2-yl) pentan-1-ol (34-3) (412 mg,1.922mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (10 mL), triphenylphosphine (1008.48 mg, 3.254 mmol,2.0 eq) was added, carbon tetrabromide (1275.08 mg, 3.254 mmol,2.0 eq) was slowly added under ice-bath, and stirred at room temperature (18 ℃ C.) for 16hr; TLC (petroleum ether: ethyl acetate=10:1) showed complete reaction; the reaction mixture was extracted with water (15 mL), ethyl acetate (10 mL x 3) and the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) and chromatographed on silica gel (petroleum ether: ethyl acetate=98:2) to give 2- (5-bromopentyl) naphthalene (34-4) (400 mg,1 443mmol, 85.32%) as a white solid. LC-MS (ESI) [ m+h ] + = 278.0.
The fifth step compound 2- (5-bromopentyl) naphthalene (34-4) (400 mg, 1.013 mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (5 mL), phthalimide (320.73 mg,1.732mmol,1.2 eq) was added and stirred at room temperature for 16hr; the reaction solution was filtered by LC-MS monitoring that no raw material remained, the filtrate was dried by spin-drying under reduced pressure (water pump, 45 ℃ C.) and then dried by oil pump to give the product 2- [5- (naphthalen-2-yl) pentyl ] isoindole-1, 3-dione (34-5) (160 mg,0.745mmol, 90.00%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =344.4.
The sixth step of the compound 2- [5- (naphthalen-2-yl) pentyl ] isoindole-1, 3-dione (34-5) (470 mg,1.369mmol,1.0 eq) was added to a reaction flask containing ethanol (5 mL), and after adding hydrazine hydrate (0.333 mL,6.843mmol,5.0 eq), stirring was carried out at 80℃for 2hr, and a large amount of white floccules precipitated; the reaction solution was filtered through celite, washed, the organic phases were combined, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give 5- (naphthalen-2-yl) pentan-1-amine (34-6) (262 mg,0.983mmol, 71.8%) as a yellow solid, the crude was directly subjected to the next reaction. LC-MS (ESI) [ m+h ] + =214.
The seventh step of compound 5- (naphthalen-2-yl) pentan-1-amine (34-6) (262 mg,1.228mmol,1.3 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (10 mL), O- (7-azabenzotriazol-1-yl) -N, N, N ', N' -tetramethylurea hexafluorophosphate (431.08 mg,1.134mmol,1.2 eq), N, N-diisopropylethylamine (0.312 mL, 1.188 mmol,2.0 eq) was added and stirred for 30min after adding 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionic acid (I) (423.98 mg,0.945mmol,1.0 eq) and stirred at room temperature for 16hr; TLC (petroleum ether: ethyl acetate=10:1) monitored the reaction; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃); the crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=90:10) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [5- (naphthalen-2-yl) pentyl ] propionamide (34-7) (100 mg,0.124mmol, 13.14%) as a white solid. LC-MS (ESI) [ m+h ] + =645.
The eighth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [5- (naphthalen-2-yl) pentyl ] propanamide (34-7) (100 mg,0.155mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (5 mL), tetrabutylammonium fluoride in tetrahydrofuran (1.445 mL,1.445mmol,10 eq) was added and the mixture was warmed to 50℃and stirred for 16hr; TLC (dichloromethane: methanol=20:1) monitored the reaction was essentially complete; the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by column chromatography on silica gel (petroleum ether: ethyl acetate=90:10 to 80:20), separating to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [5- (naphthalen-2-yl) pentyl ] propanamide (34) (25.00 mg,0.038mmol, 24.31%) as a white solid .1H NMR(400MHz,CDCl3)δ7.82–7.74(m,3H),7.60(s,1H),7.43(pd,J＝6.9,1.4Hz,2H),7.32(dd,J＝8.4,1.6Hz,1H),5.42(s,1H),3.28–3.17(m,3H),2.77(t,J＝7.6Hz,2H),2.47–2.35(m,1H),2.31–2.09(m,3H),2.04–1.89(m,3H),1.80–1.63(m,7H),1.61(s,3H),1.52(dd,J＝14.7,7.3Hz,3H),1.38(dt,J＝12.7,6.0Hz,3H),1.32–1.24(m,1H),1.09–1.00(m,2H),0.97(d,J＝6.1Hz,3H),0.91–0.83(m,2H),0.80(t,J＝6.8Hz,6H).13C NMR(101MHz,CDCl3)δ172.86,139.87,133.59,131.95,131.87,128.47,127.83,127.59,127.37,127.30,126.33,125.89,125.09,79.04,77.33,77.22,77.02,76.70,54.26,53.91,39.42,38.81,37.86,36.91,36.49,35.90,35.56,33.55,32.08,30.90,29.58,28.04,27.61,26.51,25.11,21.88,21.78,19.57,15.36,14.28.LC-MS(ESI)[M+H]+＝530.
Example 35
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3- { [3- (pyridin-2-yl) propyl ] oxy } propyl) propanamide
Sodium hydride (583.18 mg,14.579mmol,4.0 eq) as the first step compound was added to a reaction flask containing anhydrous DMF (20 mL), 2-propanol pyridine (50 mg, 3.640 mmol,1.0 eq) was added dropwise under nitrogen protection and stirred for 0.5hr, and N-Boc-3-aminopropyl bromide (867.95 mg,3.645mmol,1.0 eq) was added and stirred at room temperature (20deg.C) for 18hr; LC-MS monitored the reaction. The reaction was quenched by dropwise addition of water (10 mL), extracted with ethyl acetate (15 mL x 3), the organic phases combined, washed (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate dried under reduced pressure (water pump, 40 ℃), to give a yellow liquid; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0-10:1) and the product at rf=0.5 was collected to give the product tert-butyl (3- (3- (pyridin-2-yl) propoxy) propyl) carbamate (35-1) (200 mg,0.611mmol, 16.78%) as a yellow oil. LC-MS (ESI) [ m+h ] + = 295.25
The second step, tert-butyl (3- (3- (pyridin-2-yl) propoxy) propyl) carbamate (35-1) (100.00 mg,0.340 mmol) was added to a reaction flask containing anhydrous dichloromethane (5 mL) and TFA (46.48 mg,0.408 mmol) was added dropwise under nitrogen and stirred at 0deg.C for 2hr; TLC (dichloromethane: methanol=10:1) monitored the completion of the reaction and the reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give the product 3- [3- (pyridin-2-yl) propoxy ] propyl-1-amine (35-2) (55 mg,0.255mmol, 75.01%) as a yellow oil. LC-MS (ESI) [ m+h ] + = 195.20
The third step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (115.33 mg, 0.255 mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (5 mL), HATU (146.78 mg,0.386mmol,1.2 eq) and N, N-diisopropylethylamine (0.128 mL,0.772mmol,2.0 eq) were added and stirring was continued for 0.5hr at room temperature (20 ℃) followed by 3- [3- (pyridin-2-yl) propoxy ] propyl-1-amine (35-2) (50 mg, 0.255 mmol,1.0 eq). TLC (Petroleum ether: ethyl acetate=1:1) monitoring of the reaction showed the disappearance of starting material, with the formation of dominant sites of reduced polarity; the reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 2), the organic phases were combined, washed with water (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-9:1) to give 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -N-3- [3- (pyridin-2-yl) propoxy ] propyl ] propionamide (35-3) (52 mg,0.076mmol, 29.13%) as a colorless oil .1H NMR(400MHz,CDCl3)δ8.49(d,J＝3.9Hz,1H),7.57(m,1H),7.13(d,J＝7.8Hz,2H),6.32(d,J＝5.1Hz,1H),3.45(dt,J＝12.6,6.0Hz,4H),2.87(m,5H),1.79(m,22H),1.06(m,6H),0.85(m,12H),0.73(m,6H),-0.01(t,J＝7.4Hz,6H).
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-3- [3- (pyridin-2-yl) propoxy ] propyl ] propanamide (35-3) (50 mg,0.080mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.800 mL,0.800mmol,10.0 eq) was added and stirred at 50℃for 18hr. The reaction was monitored by TLC (dichloromethane: methanol=10:1). The reaction solution was dried under reduced pressure, ethyl acetate (10 mL) and water (20 mL) were added and mixed well, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) successively to give crude product, which was purified by prep-TLC (dichloromethane: methanol=10:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3- { [3- (pyridin-2-yl) propyl ] oxy } propyl) propionamide (35) (4 mg, 0.0070 mmol, 8.81%) as a white solid .1H NMR(400MHz,CDCl3)δ8.51(t,J＝5.6Hz,1H),7.60(td,J＝7.6,1.7Hz,1H),7.21–7.05(m,2H),6.35(s,1H),3.45(m,5H),3.21(dd,J＝11.1,3.7Hz,1H),2.94–2.79(m,2H),2.47–2.21(m,3H),2.18–2.08(m,2H),2.01(dd,J＝13.6,6.7Hz,2H),1.91(d,J＝15.4Hz,2H),1.76(dd,J＝12.0,6.3Hz,5H),1.70–1.56(m,6H),1.33(m,4H),1.05–1.00(m,2H),1.00–0.92(m,3H),0.86(d,J＝15.3Hz,1H),0.79(dd,J＝13.1,8.2Hz,6H).13C NMR(101MHz,CDCl3)δ172.89,161.53,149.23,136.43,131.93,128.23,122.87,121.15,79.05,77.35,77.24,77.04,76.72,70.08,69.68,54.25,53.91,38.80,38.07,37.97,36.92,36.48,35.80,34.78,33.50,32.08,29.76,29.10,28.05,27.61,25.27,21.86,21.77,21.40,19.53,15.38,14.26,0.01.LC-MS(ESI)[M+H]+＝511.40.
Example 36
Preparation of the Compound 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindol-5-yl) propanamide
The first step compound 1-methylindole-5-amine (42.39 mg,0.290mmol,1.2 eq) was added to a reaction flask containing N, N-dimethylformamide) (10 mL), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (101.77 mg,0.268mmol,1.2 eq), N, N-diisopropylethylamine (0.074 mL, 0.225 mmol,2.0 eq) was added, and after stirring at room temperature for 30min, 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] propionic acid (I) (100 mg,0.223mmol,1.0 eq) was added and stirred at room temperature for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; ethyl acetate (20 mL) was added to the reaction mixture, which was washed with water (20 mL. Times.2), dried under reduced pressure (water pump, 40 ℃ C.) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=90:10 to 75:25) to collect the product 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindol-5-yl) propanamide (36-1) (108 mg,0.150mmol, 67.14%) as a white solid ;1H NMR(400MHz,CDCl3)δ7.79(s,1H),7.24(s,3H),7.04(s,1H),3.78(s,3H),3.19(dd,J＝11.1,4.7Hz,1H),2.05(s,4H),1.64(s,20H),0.89(s,16H),0.66(d,J＝8.8Hz,2H),0.03(t,J＝4.3Hz,7H).
The second step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylindol-5-yl) propionamide (36-1) (108 mg, 0.87 mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), tetrabutylammonium fluoride (1.872 mL,1.872mmol,10.0 eq) was slowly added dropwise and stirred at 80℃for 18hr. LC-MS shows that after the reaction is completed, ethyl acetate (20 mL) is added to the reaction solution, the mixture is washed with water (20 mL. Times.2), dried under reduced pressure (water pump, 40 ℃ C.) to obtain a crude product, which is separated and purified by column chromatography (petroleum ether: ethyl acetate=95:5 to 90:10), and the product 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindol-5-yl) propanamide is collected (36)(45mg,0.078mmol,41.57％).1H NMR(400MHz,CDCl3)δ7.79(s,1H),7.25(d,J＝5.2Hz,2H),7.04(d,J＝3.0Hz,1H),6.43(d,J＝3.0Hz,1H),3.77(s,3H),3.23(dd,J＝11.5,4.6Hz,1H),2.68–2.53(m,1H),2.45(ddd,J＝14.1,9.4,5.0Hz,2H),2.39–2.30(m,1H),2.28–2.17(m,1H),2.00(t,J＝14.5Hz,3H),1.88–1.70(m,4H),1.68(s,3H),1.59(dd,J＝14.0,10.5Hz,3H),1.08(ddd,J＝12.9,10.1,5.4Hz,3H),0.99–0.95(m,3H),0.86–0.81(m,3H),0.80(d,J＝5.0Hz,3H).13C NMR(101MHz,CDCl3)δ171.07,134.15,131.87,130.28,129.64,128.98,128.51,115.69,112.53,109.27,100.99,79.06,77.33,77.22,77.02,76.70,54.27,53.96,38.82,37.89,36.93,36.52,36.39,33.63,32.93,32.16,28.04,27.61,27.42,25.10,21.90,21.84,19.67,15.36,14.30.LC-MS(ESI)[M+H]+＝463.3.
Example 37
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxaazepin-7-yl) butanamide
In the first step, 7-nitro-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazapine (300 mg,1.665 mmol) was dissolved in N, N-dimethylformamide (10 mL), then the mixture was cooled to 0℃under nitrogen, sodium hydride (79.93 mg,3.330 mmol) was added to the solution, stirring was continued for half an hour at 0℃and methyl iodide (0.207 mL,3.330 mmol) was added dropwise, and the mixture was warmed to room temperature and stirred for two hours. After the reaction was completed, 30mL of water was added, extracted with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and spin-dried to obtain a crude product, which was purified by a preparation plate (developing solvent: petroleum ether: ethyl acetate=5/1) to obtain 4-methyl-7-nitro-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazahexen as a pale yellow solid (37-1)(100mg,0.489mmol,29.38％).1H NMR(400MHz,CDCl3)δ7.82(dd,J＝9.0,2.6Hz,1H),7.66(d,J＝2.6Hz,1H),6.56(d,J＝9.0Hz,1H),4.27(dd,J＝5.7,3.4Hz,2H),3.48–3.44(m,2H),3.05(s,3H).
In the second step, 4-methyl-7-nitro-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazapine (37-1) (100 mg, 0.515mmol) was dissolved in ethanol (2 mL) and ethyl acetate (2 mL), palladium on carbon (30 mg) was added and then stirred overnight at room temperature under hydrogen. After the reaction was completed, the reaction solution was filtered and washed with methanol. The filtrate was dried by spinning to obtain crude product. Purifying the crude product by a preparation plate (developing agent: petroleum ether: ethyl acetate=2/1) to obtain yellow solid 4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazacyclohexan-7-amine (37-2)(30mg,0.183mmol,35.48％).1H NMR(400MHz,CDCl3)δ6.56(d,J＝8.3Hz,1H),6.25(d,J＝14.1Hz,2H),4.31–4.26(m,2H),3.14(s,2H),2.79(s,3H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (37-2) (50 mg,0.108mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (5 mL), HATU (61.63 mg,0.163mmol,1.5 eq) and N, N-diisopropylethylamine (0.05 25mL,0.325mmol,3.0 eq) were added and stirred at room temperature (20 ℃) for 0.5hr, and 4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazacyclo-hexan-7-amine (21.13 mg, 1.13 mmol, 18.2 eq) was added and stirred further. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxaazepin-7-yl) butanamide (37-3) (40 mg,0.059mmol, 54.63%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 477.30
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazazin-7-yl) butyramide (37-3) (40 mg,0.066mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), tetrabutylammonium fluoride (171.74 mg,0.657mmol,10.0 eq) was added and the mixture was stirred at 50℃for 18hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by preparative TLC (dichloromethane: methanol=10:1), separating to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (4-methyl-3, 4-dihydro-2H-benzo [2,1-b ] [1,4] oxazacyclohexan-7-yl) butyramide (37) (4 mg, 0.0070 mmol, 11.16%) as a white solid .1H NMR(400MHz,CDCl3)δ6.94(m,2H),6.65(d,J＝8.6Hz,1H),4.26(m,2H),3.17(m,3H),2.83(s,3H),2.28(td,J＝7.2,2.5Hz,2H),2.17(ddd,J＝27.8,12.8,7.1Hz,3H),2.01(m,2H),1.90(t,J＝12.6Hz,1H),1.73(m,4H),1.61(s,3H),1.39(ddd,J＝10.9,9.7,3.6Hz,2H),1.30(s,4H),1.08(m,3H),0.95(s,2H),0.89(dd,J＝12.3,5.5Hz,2H),0.85(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δδ172.80,132.42,127.17,99.99,78.39,64.80,54.46,54.23,38.52,37.78,36.92,36.22,33.25,32.07,29.32,27.95,27.25,26.93,24.34,21.70,21.60,18.39,14.73,13.39.LC-MS(ESI)[M+H]+＝495.30.
Example 38
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propanamide
The first step compound [ (hexahydropyridin-4-ylmethyl) amino ] methanoic acid-2-methylpropan-2-yl ester (500 mg,2.333mmol,1.0 eq) 2- (2-bromoethyl) pyridine (249.90 mg,1.358mmol,1.3 eq) was added to a reaction flask containing acetonitrile (5 mL), potassium carbonate (370 mg,3.50mmol,1.5 eq) was added and stirred at room temperature (25 ℃ C.) under nitrogen for 16hr. TLC (dichloromethane: methanol=20:1) monitored the reaction. After the reaction solution was added to the aqueous solution, dichloromethane (20 ml. Times.3) was added to extract, the organic phases were combined, dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (dichloromethane: methanol=1:0 to 9:1) to give the product [ ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) amino ] methanoic acid-2-methylpropan-2-yl ester (38-1) (270 mg,0.884mmol, 37.89%) as a pale yellow oil, LC-MS (ESI) [ m+h ] + = 356.82
The second step compound [ ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) amino ] methanoic acid-2-methylpropan-2-yl ester (38-1) (100 mg, 0.227 mmol,1.0 eq) was added to a reaction flask containing methanol (1 mL), and after adding a methanol solution of hydrogen chloride (0.819 mL,3.274mmol,10eq, 4M), stirred at room temperature (25 ℃ C.) for 16hr. LC-MS monitored the reaction. The reaction solution was directly dried under reduced pressure (water pump, 50 ℃ C.) and then was further dried by an oil pump (45 ℃ C., water bath) to give {1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methanamine (38-2) (80 mg,0.298mmol, 91.07%) as a yellow oil. LC-MS (ESI) [ m+h ] + =202.2
The third step compound {1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methanamine (38-2) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.074 mL, 0.4476 mmol,2.0 eq) were added and stirring was continued for 1hr after stirring at room temperature (25 ℃ C.) for 0.5hr, followed by addition of [1- (pyridin-2-ylmethyl) hexahydropyridin-4-yl ] methanamine (54.90 mg,0.267mmol,1.2 eq); TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) to give the crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propionamide (38-3) (120 mg,0.151mmol, 67.73%) as a yellow oil, which was directly added to the new point.
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propanamide (38-3) (110 mg,0.173mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.729 mL,1.729mmol,10 mmol, 1M) was added and then stirred at 50℃for 16hr; TLC (dichloromethane: methanol=10:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 45 ℃) and then added with ethyl acetate (20 mL) and water (20 mL) to dissolve the mixture, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 4) and dried under reduced pressure (water pump, 45 ℃) to give crude product, which was purified by prep-TLC (dichloromethane: methanol=10:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- ({ 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } methyl) propionamide (38) (32.05 mg,0.059mmol, 34.13%) as a white solid .1H NMR(400MHz,CDCl3)δ8.51(dd,J＝4.9,0.8Hz,1H),7.60(td,J＝7.7,1.8Hz,1H),7.19(d,J＝7.8Hz,1H),7.14–7.08(m,1H),5.66(s,1H),3.36(dd,J＝10.1,7.0Hz,2H),3.28–3.19(m,1H),3.18–3.10(m,2H),3.10–2.99(m,4H),2.80(dd,J＝9.6,6.4Hz,2H),2.45(ddd,J＝13.7,9.5,7.2Hz,1H),2.40–2.19(m,4H),2.19–2.05(m,5H),2.03–1.85(m,4H),1.79–1.71(m,3H),1.66(d,J＝1.4Hz,3H),1.63(s,3H),1.56(dd,J＝12.7,8.8Hz,2H),1.45(dt,J＝9.4,4.7Hz,2H),1.41–1.32(m,3H),1.05(dd,J＝18.6,5.5Hz,5H),1.00–0.94(s,3H),0.91–0.85(m,2H),0.82(dd,J＝13.8,6.8Hz,6H).13C NMR(101MHz,CDCl3)δ173.08,160.17,149.23,136.45,131.81,128.53,123.27,121.27,79.01,59.03,58.52,54.28,53.91,53.26,44.91,38.82,37.83,36.92,36.50,35.81,35.60,35.51,33.57,32.11,29.70,28.06,27.62,25.11,24.15,21.90,21.76,19.79,19.60,15.39,14.29,13.70.LC-MS(ESI)[M+H]+＝536.4.
Example 39
Preparation of the Compound N- (7-cyclohexylheptyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionamide
In the first step, methyl 6-bromohexanoate (5.0 g,23.9mmol,1.0 eq) and triphenylphosphine (18.8 g,71.7mmol,3.0 eq) were dispersed in acetonitrile (20 mL) and the reaction was stirred overnight at 90 ℃. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. After the reaction mixture was concentrated, the crude product was purified by passing through a column with methylene chloride: methanol=20:1, and after concentration, methyl 6- (bromotriphenyl-. Lamda. 5 -methyl) hexanoate (39-1) (8.6 g, purity 90%, yield 68.7%) was obtained. LC-MS (ESI) [ m+h ] + =391.2.
In the second step, potassium tert-butoxide (1.0 g,8.9mmol,2.0 eq) was dispersed in acetonitrile (20 mL), methyl 6- (bromotriphenyl-. Lamda. 5 -methyl) hexanoate (39-1) (4.2 g,8.9mmol,2.0 eq) was added under ice-bath, and the reaction solution was stirred at 90℃overnight. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (100 mL), and washed with water (50 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with methylene chloride: methanol=20:1, and concentrated to give methyl (6E) -7-cyclohexylhept-6-enoate (39-2) (0.7 g, purity 90%, yield 63％).1H NMR(400MHz,CDCl3)δ5.26–5.19(m,2H),3.67(s,3H),2.32(t,J＝7.5Hz,2H),2.25–2.18(m,1H),2.05(dd,J＝13.0,7.3Hz,2H),1.65(ddd,J＝27.7,16.0,10.9Hz,8H),1.39–1.35(m,2H),1.29–1.22(m,3H),1.17(ddd,J＝12.5,7.7,3.1Hz,1H),1.04(dt,J＝12.2,9.5Hz,2H).
In the third step LiAlH 4 (lithium aluminum hydride) (0.24 g,6.2mmol,2.0 eq) was dispersed in THF (tetrahydrofuran) (200 mL), and a solution of methyl (6E) -7-cyclohexylhept-6-enoate (39-2) (0.7 g,3.1mmol,1.0 eq) in THF (tetrahydrofuran) (50 mL) was added while cooling with ice and the reaction mixture was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give (6E) -7-cyclohexylhept-6-en-1-ol (39-3) (0.6 g, purity 85%, yield 83.25%), the crude was directly fed to the next step.
In the fourth step, (6E) -7-cyclohexylhept-6-en-1-ol (39-3) (0.45 g,2.3mmol,1.0 eq) was dissolved in THF (tetrahydrofuran) solution (25 mL) and palladium on carbon catalyst (90 mg) was added under nitrogen. The reaction solution was replaced with hydrogen 3 times. The reaction mixture was stirred at room temperature for 16hrs under hydrogen atmosphere. 1 H NMR showed the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give 7-cyclohexylhept-1-ol (39-4) (0.6 g, purity 85%, yield) 83.25％).1H NMR(400MHz,CDCl3)δ3.64(t,J＝6.6Hz,2H),1.70(d,J＝1.9Hz,2H),1.67(d,J＝1.9Hz,3H),1.61(dd,J＝7.3,2.8Hz,1H),1.59–1.54(m,2H),1.36–1.31(m,3H),1.28–1.25(m,5H),1.23–1.19(m,2H),1.18–1.10(m,5H),0.86(dt,J＝21.1,6.4Hz,3H).
In the fifth step, 7-cyclohexylhept-1-ol (39-4) (0.5 g,2.5mmol,1.0 eq) was dissolved in methylene chloride (50 mL), then triphenylphosphine (1.32 g,5.0mmol,2.0 eq) and carbon tetrabromide (1.67 g,5.0mmol,2.0 eq) were added and the reaction solution was stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=20:1) showed that the reaction was complete. After termination of the reaction, the reaction mixture was washed with water (40 mL. Times.2), the organic layer was dried over anhydrous sodium sulfate, and the crude product was concentrated and purified by petroleum ether: ethyl acetate=20:1, to give (7-bromoheptyl) cyclohexane (39-5) (0.45 g, purity 90%, yield 63.4％).1H NMR(400MHz,CDCl3)δ3.41(t,J＝6.9Hz,2H),1.91–1.80(m,2H),1.66(t,J＝15.2Hz,6H),1.41(dd,J＝14.0,6.4Hz,2H),1.32–1.22(m,8H),1.16(dd,J＝18.5,13.5Hz,6H),0.90–0.77(m,3H).
In the sixth step, (7-Bromoheptyl) cyclohexane (39-5) (0.45 g,1.7mmol,1.0 eq) and potassium phthalimide (1.38 g,2.1mmol,1.2 eq) were dissolved in DMF (N, N-dimethylformamide) (5 mL) and the reaction mixture was stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (25 mL. Times.2), and washed with water (30 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was purified by passing through a column with petroleum ether ethyl acetate=5:1 to give 2- (7-cyclohexylheptyl) isoindole-1, 3-dione (39-6) (0.4, purity 95%, yield 67.3%). LC-MS (ESI) [ m+na ] +＝350.2,tR =1.68 min.
In a seventh step, 2- (7-cyclohexylheptyl) isoindole-1, 3-dione (39-6) (0.15 g,0.46mmol,1.0 eq) was dissolved in ethanol (20 mL), hydrazine hydrate (0.30 mL,2.3mmol,5.0 eq) was added, the reaction was stirred at 80℃for 3h, and TLC (dichloromethane: methanol=10:1) showed completion of the reaction. After the completion of the reaction, the mixture was concentrated, extracted with chloroform (20 mL), washed with water (20 mL. Times.2), and dried over anhydrous Na 2SO4 to give crude 7-cyclohexylhept-1-amine (39-7) which was directly taken into the next step.
In the eighth step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min, the reaction 7-cyclohexylhept-1-amine (39-7) (52.8 mg,0.29mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 10:1 to give N- (7-cyclohexylheptyl) -3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (39-8) which was taken directly into the next step.
The crude N- (7-cyclohexylheptyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (39-8) (60 mg,0.09mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), a solution of TBAF (1.12 mL,1.12mmol,10.0 eq) in tetrahydrofuran was added, and the reaction mixture was stirred overnight at 60 ℃. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=5:1 was passed through the column, after concentration N- (7-cyclohexylheptyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide N- (7-cyclohexylheptyl) is obtained 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (39) (12.8 mg, purity 98%, yield 20.6％).1H NMR(400MHz,CDCl3)δ5.48(s,1H),3.24-3.20(m,3H),2.49–2.10(m,5H),2.05–1.86(m,3H),1.77(dt,J＝13.1,3.4Hz,1H),1.69(s,3H),1.66(s,4H),1.64–1.55(m,5H),1.50-1.45(m,2H),1.41–1.34(m,1H),1.33–1.24(m,8H),1.23–1.18(m,2H),1.16-1.13(m,4H),1.11–0.93(m,6H),0.93-0.83(m,4H),0.81-0.79(m,6H).13C NMR(101MHz,CDCl3)δ172.87,131.92,128.50,79.05,77.35,77.23,77.03,76.71,54.30,53.95,39.57,38.83,37.88,37.67,37.51,36.94,36.51,35.61,33.58,33.46,32.12,29.87,29.69,29.35,28.05,27.63,26.96,26.79,26.77,26.46,25.15,21.91,21.80,19.59,15.38,14.30.LC-MS(ESI)[M+1]+＝514.6(Ms+1)
Example 40
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindazol-5-yl) butanamide
The first step compound 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (90 mg,0.194mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (2- (7-azobenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate) (88.73 mg,0.233mmol,1.2 eq) and N, N-diisopropylethylamine (0.064 mL,0.389mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by 1-methylindazole-5-amine (37.21 mg,0.25 mmol, 1.25 hr). TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 8:2) to separate 4- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -N- (1-methylindol-5-yl) butanamide (40-1) (70 mg,0.095mmol, 50.07%) as a white solid .1H NMR(400MHz,CDCl3)δ7.99(d,J＝22.9Hz,2H),7.42(d,J＝8.5Hz,1H),7.16(s,1H),4.09(s,3H),3.18(dd,J＝11.0,4.7Hz,1H),2.37(dd,J＝14.9,8.1Hz,2H),2.21(dd,J＝21.1,13.3Hz,3H),2.01(d,J＝5.6Hz,2H),1.62(s,30H),1.26(s,8H),0.94–0.77(m,3H).
The second step compound 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthryl ] -N- (1-methylindazol-5-yl) butanamide (40-1) (70 mg,0.118mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.183 mL,1.183mmol,10 eq) was added and then warmed to 50℃and stirred for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a brown oil, water (20 mL) was added, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 7:3) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindole-5-yl) butyramide (40) (35 mg,0.062mmol, 52.78%) as a white solid .1H NMR(400MHz,CDCl3)δ7.98(s,1H),7.93(s,1H),7.37(dt,J＝24.4,5.3Hz,2H),7.20(s,1H),4.06(s,3H),3.22(dd,J＝11.5,4.6Hz,1H),2.42–2.31(m,2H),2.22–2.08(m,3H),2.04–1.81(m,4H),1.77–1.64(m,5H),1.62(s,3H),1.59-1.43(m,3H),1.42–1.25(m,2H),1.10–1.02(m,2H),0.97(s,3H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ171.35,137.48,132.69,132.42,127.80,127.52,124.05,121.09,111.91,109.21,79.11,54.30,53.97,38.83,37.85,37.50,36.97,36.53,35.64,33.59,32.13,28.52,28.06,27.65,24.42,21.86,19.68,15.38,14.30.LC-MS[M+H]+＝478.3
Example 41
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (naphthalen-2-yl) butanamide
The first step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (90 mg,0.194mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate) (88.73 mg,0.233mmol,1.2 eq) and N, N-diisopropylethylamine (0.064 mL,0.389mmol,2.0 eq) were added, and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by naphthalene-2-oxamine (20 mg,0.25 mmol,1.3 hr). TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 8:2) to give the product 4- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (naphthalen-2-yl) butanamide (41-1) (83 mg,0.113mmol, 80%) as a white solid .1H NMR(400MHz,CDCl3)δ8.21(s,1H),7.79(d,J＝8.3Hz,3H),7.45(d,J＝7.8Hz,3H),3.18(dd,J＝11.2,4.6Hz,1H),2.38(s,2H),2.16(d,J＝8.3Hz,2H),2.04(s,4H),1.90(d,J＝12.2Hz,3H),1.69(d,J＝3.8Hz,3H),1.63(s,6H),1.54(s,9H),1.37(d,J＝15.9Hz,1H),1.26(t,J＝7.1Hz,5H),1.10–1.01(m,2H),0.99(s,0H),0.94(s,1H),0.84–0.78(m,5H),0.76(d,J＝6.3Hz,4H).
The second step compound 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (naphthalen-2-yl) butanamide (41-1) (90 mg,0.156mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.560 mL, 1.560 mmol,10 eq) was added and then stirred at 50℃for 16hr; TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a brown oil, which was added with water (20 mL), extracted with ethyl acetate (20 ml×3), the organic phases were combined, washed with water (30 ml×2), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:2) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (naphthalen-2-yl) butanamide (41) (47 mg,0.079mmol, 50.7%) as a white solid .1H NMR(400MHz,CDCl3)δ8.21(s,1H),7.82–7.75(m,3H),7.43(m,3H),3.22(dd,J＝11.5,4.7Hz,1H),2.39(dd,J＝14.9,7.7Hz,2H),2.17(t,J＝8.3Hz,3H),2.00(s,1H),1.99(s,1H),1.90(d,J＝15.0Hz,2H),1.76(dd,J＝13.0,3.4Hz,1H),1.73–1.66(m,3H),1.62(s,4H),1.57(dd,J＝12.8,3.5Hz,2H),1.43–1.24(m,2H),1.14–1.00(m,2H),0.97(d,J＝5.6Hz,3H),0.83–0.77(m,7H).13C NMR(101MHz,CDCl3)δ171.39,141.69,135.36,133.89,132.39,130.60,128.77,127.85,127.65,127.55,126.52,124.97,119.75,116.47,79.12,54.30,53.97,38.84,37.84,36.97,36.54,33.59,32.14,28.49,28.06,27.65,24.32,21.94,21.86,19.69,15.38,14.30.LC-MS[M+H]+＝474.3.
Example 42
Preparation of the Compound N- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } -N- [7- (pyridin-2-yl) heptyl ] glycine
In the first step, 2- (7-bromoheptyl) pyridine (III-2) (0.3 g,1.17mmol,1.0 eq) was dispersed in ethanol (10 mL), and triethylamine (0.65 mL,4.68mmol,4.0 eq) and glycine methyl ester hydrochloride (0.21 g,2.34mmol,2.0 eq) were added at room temperature and the reaction stirred at 90℃for 48h. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. After the reaction mixture was concentrated, the crude product was extracted with ethyl acetate (100 mL) and washed with water (50 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with methylene chloride: methanol=20:1, and concentrated to give ethyl { [7- (pyridin-2-yl) heptyl ] amino } acetate (42-1) (0.11 g, purity 90%, yield 30.4%). LC-MS (ESI) [ m+h ] +＝279.2,tR =0.22 min.
In a second step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min the reaction { [7- (pyridin-2-yl) heptyl ] amino } acetic acid ethyl ester (42-1) (66.54 mg,0.29mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product purified with dichloromethane: methanol=15:1 was directly taken to the next step.
The crude product of the previous step (80 mg,0.11mmol,1.0 eq) was dissolved in methanol (5 mL), and a 20% aqueous NaOH solution (0.5 mL) was added under ice-bath, and the reaction was stirred at room temperature for 2h. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (30 mL), washed with saturated ammonium chloride (20 mL) and washed with water (20 mL). The organic layer was dried over anhydrous sodium sulfate and the crude product was purified by passing through a column with methylene chloride methanol=10:1 to give N- {3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } -N- [7- (pyridin-2-yl) heptyl ] glycine (42) (10.0 mg, 92% purity, yield 12.0％).1H NMR(400MHz,DMSO)δ8.74(t,J＝5.1Hz,1H),8.31(dt,J＝15.3,7.8Hz,1H),7.76(ddd,J＝19.3,17.1,7.3Hz,2H),4.07(s,1H),3.89(s,1H),3.32–3.18(m,2H),3.00-2.95(m,1H),2.92(dd,J＝15.2,7.3Hz,2H),2.40–2.11(m,4H),2.11–1.73(m,5H),1.64-1.55(m,5H),1.57–1.34(m,7H),1.30-1.10(m,8H),1.09–0.91(m,3H),0.90–0.79(m,4H),0.78–0.71(m,4H),0.71–0.62(m,3H).LC-MS(ESI)[M+1]+＝567.4(Ms+1).
Example 43
Preparation of the Compound N- [6- (3-fluorophenyl) hexyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
The first step compound methyl 5-bromopentanoate (7.715 g,39.55mmol,1.0 eq) was added to a reaction flask containing acetonitrile (100 mL), triphenylphosphine (31.12 g,262.29mmol,3 eq) was added and stirred at 80℃for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; ethyl acetate (200 mL) was added to the reaction solution, and after washing with water (200 mL x 2), the mixture was dried under reduced pressure (pump, 40 ℃) to give a crude product, which was separated and purified by column chromatography (dichloromethane: methanol=1:0 to 100:1), and methyl 5- (bromotriphenyl-. Lamda.5-methylphospholate) (43-1) (9.80 g,20.354mmol, 51.47%) was collected as a colorless oily liquid; LC-MS (ESI) [ m+h ] + = 337.3.
In a second step, 3-fluorobenzene-1-carbaldehyde (2 g,16.11mmol,1.0 eq) was added to a reaction flask containing acetonitrile (20 mL), methyl 5- (bromotriphenyl-. Lamda.5-methyl) valerate (43-1) (14.74 g,32.23mmol,2.0 eq) was added after hydrogen was replaced, and potassium carbonate (4.45 g,32.23mmol,2.0 eq) was added and stirred at 90℃for 16hr. LC-MS showed that after completion of the reaction, ethyl acetate (200 mL) was added, washed with water (200 mL. Times.2), and dried under reduced pressure (water pump, 40 ℃ C.) to give the crude product as a colorless oil by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) separation and purification of methyl (5E) -6- (3-fluorophenyl) hex-5-enoate (43-2) (1.6 g,5.759mmol, 35.74%). LC-MS (ESI) [ m+h ] + = 223.11.
The third step compound (5E) -6- (3-fluorophenyl) hex-5-enoic acid methyl ester (43-2) (1.6 mg,5.759mmol,1.0 eq) was added to a reaction flask containing methanol (5 mL), after hydrogen was replaced, palladium on carbon (150 mg) was added and then stirred for 12hr. LC-MS showed that after the reaction was complete, the celite was filtered and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give the product methyl 6- (3-fluorophenyl) hexanoate (43-3) (1.6 g,5.707mmol, 79.28%) as a colorless oil. LC-MS (ESI) [ m+h ] + = 225.12.
The fourth step compound, methyl 6- (3-fluorophenyl) hexanoate (43-3) (1.6 g,5.707mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (20 mL), lithium aluminum hydride (0.29 g,5.707mmol,1 eq) was added at 0deg.C, and then stirred at room temperature (18deg.C) for 16hr; TLC (Petroleum ether: ethyl acetate=.3:1) monitored reaction was complete; the reaction solution was dried under reduced pressure (water pump, 50 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give 6- (3-fluorophenyl) hex-1-ol (43-4) (1.5 g,6.114mmol, 80.67%) as a colorless oily liquid. LC-MS (ESI) [ m+h ] + = 179.13.
The fifth step compound 1- (6-bromohexyl) -3-fluorobenzene (43-4) (1.2 g,6.114mmol,1.0 eq) was added to a reaction flask containing methylene chloride (10 mL), triphenylphosphine (2.95 g,9.172mmol,1.5 eq) was added, and carbon tetrabromide (2.41 g,9.172mmol,1.5 eq) was stirred at room temperature (18 ℃ C.) for 16hr; TLC (petroleum ether: ethyl acetate=.3:1) monitored complete reaction of starting material. The reaction mixture was dried under reduced pressure (water pump, 50 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product 1- (6-bromohexyl) -3-fluorobenzene (43-5) (1.0 g,3.087mmol, 50.49%) as a white solid. LC-MS (ESI) [ m+h ] + = 259.04.
The sixth step of the compound 1- (6-bromohexyl) -3-fluorobenzene (43-5) (60 mg,0.232mmol,1.0 eq) was charged into a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), and after adding potassium phthalimide salt (43.06 mg,0.232mmol,1.2 eq), it was stirred at room temperature (18 ℃ C.) for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1) and the new spot (rf=0.55) was collected to give the product 1- (6-bromohexyl) -3-fluorobenzene (43-6) (40.0 mg,0.098mmol, 42.48%) as a colorless oil. LC-MS (ESI) [ m+h ] + = 326.15.
The seventh step of adding 1- (6-bromohexyl) -3-fluorobenzene (43-6) (40.0 mg,0.098mmol,1.0 eq) to a reaction flask containing ethanol (3 mL), adding hydrazine hydrate (0.045 mL,0.98mmol,10.0 eq) and heating to 80℃for stirring for 16hr; the reaction was monitored by LC-MS. The reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃ C.) and then was substituted with oil pump to give 6- (3-fluorophenyl) hex-1-amine (43-7) (40 mg,0.154mmol, 99.97%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =196.28.
The eighth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (160 mg, 0.356 mmol,1.0 eq)) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg, 0.67 mmol,1.2 eq) and N, N-diisopropylethylamine (0.074 mL, 0.4476 mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by 6- (3-fluorophenyl) hex-1-amine (43-7) (69.71 mg,0.357mmol,1 eq). TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (3-fluorophenyl) hexyl ] propionamide (43-8) (60 mg,0.077mmol, 14.97%) as a pale yellow oil, which was directly taken to the next step;
The ninth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (3-fluorophenyl) hexyl ] propanamide (43-8) (60 mg,0.077mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.445 mL,1.445mmol,10 eq) was added and then warmed to 50℃and stirred for 16hr; TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by prep-TLC (dichloromethane: methanol=10:1), isolated to give the product N- [6- (3-fluorophenyl) hexyl ] -3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (43) (6.00 mg,0.01 mmol, 13.21%) as a white solid .1H NMR(400MHz,CDCl3)δ7.24–7.17(m,1H),6.92(d,1H),6.89–6.80(m,2H),5.43–5.34(m,1H),3.24–3.17(m,3H),2.60–2.55(m,2H),2.45–2.37(m,1H),2.36–2.28(m,1H),2.25–2.18(m,1H),2.15–2.10(m,1H),1.99–1.87(m,3H),1.74–1.70(m,1H),1.68–1.65(m,1H),1.61(s,6H),1.57(d,2H),1.49–1.43(m,2H),1.35–1.30(m,6H),1.25(s,1H),1.06–0.98(m,2H),0.95(s,2H),0.91–0.81(m,3H),0.78(d,6H).13C NMR(101MHz,CDCl3)δ172.85,164.12,145.16,131.89,129.66,128.49,124.01,115.03,112.40,79.05,54.29,53.94,39.46,38.82,37.89,36.93,36.51,35.59,33.56,32.12,31.02,29.63,28.83,28.04,27.63,26.76,25.14,21.90,21.79,19.57,15.36,14.28.LC-MS[M+H]+＝512.4.
Example 44
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } propanamide
The first step compound, 2-bromopyridine (2000 mg,12.58mmol,1.0 eq), was added to a reaction flask containing acetonitrile (10 mL), but-3-yn-1-ol (2126 mg,18.87mmol,1.5 eq), diphenylphosphine palladium dichloride (1770 mg,2.516mmol,0.02 eq), cuprous iodide (480 mg,2.516mmol, 0.02eq), and N, N-diisopropylethylamine (2449 mg,18.87mmol,5 eq) were added and then stirred at 80 ℃ for 2hr; TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction; ethyl acetate (20 mL) was added to the reaction solution, which was washed with water (20 mL x 2), dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1), and the product 4- (pyridin-2-yl) but-3-yn-1-ol (44-1) (2306 mg,19.04mmol, 50.00%) was collected as a colorless oily liquid. LC-MS (ESI) [ m+h ] + = 148.02.
The second step compound, 4- (pyridin-2-yl) but-3-yn-1-ol (44-1) (750 mg,4.794mmol,1.0 eq), was added to a reaction flask containing methanol (5 mL), after which palladium on carbon (150 mg) was added in place of hydrogen, and then stirred for 12hr. LC-MS showed that after the reaction was complete, the celite was filtered and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give the product 7- (pyrazin-2-yl) heptan-1-ol (44-2) (400 mg,2.685mmol, 56.01%) as a colorless oil. LC-MS (ESI) [ m+h ] + =152.0.
The third step compound 4- (pyridin-2-yl) butan-1-ol (44-2) (330 mg,2.182mmol,1.0 eq) was added to a reaction flask containing dichloromethane (5 mL), after which hydrogen was replaced, dessert-Martin-oxidant (1110.77 mg, 2.612 mmol,1.2 eq) was added and stirred for 16hr. LC-MS showed that after the reaction was complete, the reaction mixture was extracted with water (15 mL), ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give the product 5- (pyridin-2-yl) valeraldehyde (44-3) (192 mg,0.941mmol, 43.12%) as a colorless oil. LC-MS (ESI) [ m+h ] + =150.0.
The fourth step compound, 5- (pyridin-2-yl) valeraldehyde (44-3) (192 mg,0.941mmol,1.0 eq), was added to a reaction flask containing tetrahydrofuran (10 mL), after displacement of nitrogen, 2-methylpropan-2-yl (hexahydropyridin-4-ylamino) methanoate (215.3 mg,2.15mmol,1.0 eq), sodium borohydride acetate (797.7 mg,3.764mmol,4.0 eq) and a drop of acetic acid were added and stirred at 25℃for 16hr. TLC (Petroleum ether: ethyl acetate=.3:1) showed that after completion of the reaction, the reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.4), dried over anhydrous sodium sulfate, filtered, and the filtrate was spun dry (water pump, 40 ℃ C.) under reduced pressure to give crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give ({ 1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } amino) methanoic acid-2-methylpropan-2-yl ester (44-4) (154.5 mg, 0.460 mmol, 49.30%) as a white solid. LC-MS (ESI) [ m+h ] + = 334.24.
The fifth step compound ({ 1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } amino) methanoic acid-2-methylpropan-2-yl ester (44-4) (80 mg,0.240mmol,1.0 eq) was added to a reaction flask containing 4mol/L of hydrochloric acid/1-4 dioxane solution (4 mL) and stirred for 4hr. TLC (petroleum ether: ethyl acetate=.3:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 50 ℃ C.) to give crude product as white solid (44-5) which was directly poured downward. LC-MS (ESI) [ m+h ] + = 234.36.
The sixth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (99.8 mg,0.223mmol,1.0 eq)) was added to a reaction flask containing DMF (N, N-dimethylformamide) (5 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-diisopropylethylamine (0.074 mL,0.446mmol,2.0 eq) were stirred at room temperature (18 ℃) for 0.5hr, and then 1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-amine (44-5) (50.00 mg,0.214mmol,0.96 eq) was added and stirring was continued for 1hr. TLC (dichloromethane: methanol=20:1) monitored the reaction; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } propionamide (44-6) (60 mg,0.0903mmol, 40.52%) as a white solid. LC-MS (ESI) [ m+h ] + = 664.5.
The seventh step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } propionamide (44-6) (60 mg,0.0903mmol, 40.52%) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.74 mL,0.740mmol,10 eq) was added and then warmed to 50℃and stirred for 16hr. TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [4- (pyridin-2-yl) butyl ] hexahydropyridin-4-yl } propionamide (44) (7.63 mg,0.0295mmol, 18.78%) as a white solid .1H NMR(400MHz,CDCl3)δ8.51(d,1H),7.59(t,1H),7.17–7.07(m,2H),5.60–5.46(m,1H),3.94–3.82(m,1H),3.27–3.04(m,4H),2.81(t,2H),2.59(t,2H),2.48–2.38(m,1H),2.35–2.26(m,3H),2.26–2.19(m,1H),2.17–2.09(m,2H),2.00–1.87(m,5H),1.78–1.60(m,14H),1.35(d,2H),1.08–0.99(m,2H),0.97(d,3H),0.88–0.83(m,4.3Hz,2H),0.80(d,6H).13C NMR(101MHz,CDCl3)δ172.44,161.49,149.22,136.46,131.75,128.71,122.86,121.18,79.04,57.68,54.28,53.93,51.99,45.46,38.83,37.76,36.93,36.50,35.42,33.58,32.10,30.87,28.05,27.63,27.36,25.32,24.96,21.89,21.81,19.64,15.38,14.30.LC-MS[M+H]+＝550.5.
Example 45
Preparation of the Compound N- [6- (2-fluorophenyl) hexyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
Reference example 43 conversion of m-fluorobenzaldehyde to o-fluorobenzaldehyde gives the compound N- [6- (2-fluorophenyl) hexyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (45).1H NMR(400MHz,CDCl3)δ7.20–7.12(m,2H),7.07–6.95(m,2H),3.26–3.19(m,3H),2.63(t,J＝7.7Hz,2H),2.49–2.38(m,1H),2.38–2.29(m,1H),2.24(ddd,J＝15.0,9.8,5.3Hz,1H),2.19–2.07(m,2H),2.05–1.84(m,3H),1.76(dt,J＝13.1,3.5Hz,1H),1.72–1.65(m,3H),1.65–1.56(m,6H),1.53–1.45(m,2H),1.40–1.30(m,4H),1.25(s,1H),1.17–0.99(m,3H),0.96(s,3H),0.91–0.83(m,2H),0.81(dd,J＝9.4,5.6Hz,6H).
13C NMR(101MHz,CDCl3)δ171.83,161.40,158.87,130.87,129.58,129.53,128.23,127.48,126.37,126.29,122.85,122.81,114.21,113.99,78.03,76.31,76.20,75.99,75.67,53.26,52.92,38.46,37.80,36.86,35.90,35.48,34.55,32.54,31.09,29.01,28.58,27.89,27.02,26.60,25.68,24.11,20.87,20.77,18.55,14.34,13.25.LC-MS(ESI)[M+1]+＝512(Ms+1).
Example 46
Preparation of the Compound 4- [ (4 aS,4bR,7S,8 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (6-phenylhexyl) butanamide
In the first step, (6-bromohexyl) benzene (0.5 g,2.07mmol,1.0 eq) and potassium phthalimide (0.46 g,2.49mmol,1.2 eq) were dissolved in DMF (N, N-dimethylformamide) (5 mL) and the reaction stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (50 mL. Times.2), and washed with water (60 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was purified by passing through a column with petroleum ether ethyl acetate=5:1 to give 2- (6-phenylhexyl) isoindole-1, 3-dione (46-1) (0.4 g, purity 90%, yield 56.3%). LC-MS (ESI) [ m+h ] + =330.1.
In the second step, 2- (6-phenylhexyl) isoindole-1, 3-dione (46-1) (0.2 g,0.65mmol,1.0 eq) was dissolved in ethanol (10 mL), hydrazine hydrate (0.15 mL,3.07mmol,5.0 eq) was added, the reaction was stirred at 80℃for 2h, and TLC (dichloromethane: methanol=10:1) showed completion of the reaction. Concentrating after the reaction, extracting with chloroform (20 mL), washing with water (20 mL. Times.2), drying with anhydrous Na 2SO4 to obtain crude 6-phenylhex-1-amine (46-2), and directly feeding into the next step .1H NMR(400MHz,MeOD)δ7.23(t,J＝7.5Hz,2H),7.19–7.08(m,3H),2.73–2.66(m,2H),2.60(t,J＝7.6Hz,2H),1.69–1.58(m,2H),1.56–1.44(m,2H),1.43–1.26(m,4H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (60 mg,0.130mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (88.73 mg,0.233 mmol) and N, N-Diisopropylethylamine (DIEA) (0.064 mL,0.389 mmol) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by addition of 6-phenylhex-1-amine (46-2) (44.82 mg,0.253 mmol) and stirring continued for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored the reaction. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 3), dried under reduced pressure (water pump, 40 ℃) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1) to give the product 4- [ (4 as,4br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -N- (6-phenylhexyl) butanamide (46-3) (70 mg,0.101mmol, 52.07%) as a pale yellow solid .1H NMR(400MHz,CDCl3)δ7.29(s,2H),7.17(t,J＝7.0Hz,3H),5.39(s,1H),3.32–3.13(m,3H),2.60(t,J＝7.6Hz,2H),2.27–2.01(m,5H),1.92(d,J＝14.4Hz,2H),1.69(d,J＝17.9Hz,8H),1.62(s,3H),1.59(s,3H),1.55–1.45(m,4H),1.35(s,4H),1.02(t,J＝14.7Hz,2H),0.94–0.84(m,13H),0.80(s,3H),0.75(d,J＝8.1Hz,3H),0.03(d,J＝2.6Hz,6H).
The fourth step compound 4- [ (4 aS,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (6-phenylhexyl) butanamide (46-3) (70 mg,0.113 mmol) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (1.125 mL,1.125 mmol), the mixture was warmed to 50℃and stirred for 16hr. The reaction was monitored by TLC (petroleum ether: ethyl acetate=1:1). The reaction mixture was dried under reduced pressure (water pump, 45 ℃ C.) to give a crude product, which was dissolved in ethyl acetate (20 mL), washed with water (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, dried under reduced pressure (water pump, 40 ℃ C.) and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) and prep-HPLC (Xb ridge-C18 150x19mm,5um M mobile phase: ACN- -H2O (0.1% FA)) to give 4- [ (4 aS,4bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (6-phenylhexyl) butyramide (46) (9.07 mg,0.017mmol, 15.41%) as a white solid .1H NMR(400MHz,CDCl3)δ7.26(t,J＝3.4Hz,2H),7.17(t,J＝7.0Hz,3H),5.37(d,J＝15.6Hz,1H),3.23(dt,J＝12.0,6.0Hz,3H),2.60(t,J＝7.7Hz,2H),2.11(m,5H),1.91(m,3H),1.75(m,3H),1.69–1.62(m,4H),1.59(s,3H),1.58–1.45(m,4H),1.38(m,5H),1.06(m,2H),0.96(s,3H),0.93–0.82(m,3H),0.79(d,J＝8.6Hz,6H).13C NMR(101MHz,CDCl3)δ172.87,142.62,132.57,128.37,128.25,127.52,125.64,79.10,77.33,77.22,77.01,76.70,54.31,53.98,39.49,38.83,37.87,36.98,36.72,36.52,35.85,33.56,32.10,31.31,29.66,28.92,28.60,28.05,27.66,26.80,24.59,21.94,21.86,19.59,15.37,14.29.LC-MS(ESI)[M+H]+＝508.4.
Example 47
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } propanamide
The first step compound 3- (pyridin-2-yl) propan-1-ol (600 mg,4.37mmol,1.0 eq) was added to a reaction flask containing acetonitrile (10 mL), triphenylphosphine (1721 mg,6.57mmol,1.5 eq) was added, and after carbon tetrabromide (2175 mg,6.57mmol,1.5 eq) was stirred at room temperature (0 ℃ C.) for 0.5hr and then allowed to stir to room temperature for 16hr; TLC (Petroleum ether: ethyl acetate=.3:1) monitoring the reaction of the raw materials, and generating a point with reduced polarity; the reaction solution was dried under reduced pressure (water pump, 50 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product 2- (2-bromoethyl) pyridine (47-1) (400 mg,2.15mmol, 49.20%) as a pale yellow liquid. LC-MS (ESI) [ m+h ] + = 191.1.
The second step compound 2- (2-bromoethyl) pyridine (47-1) (400 mg,2.15mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), after nitrogen substitution, (hexahydropyridin-4-ylamino) methanoic acid-2-methylpropan-2-yl ester (215.3 mg,2.15mmol,1.0 eq) and cesium carbonate (698 mg,8.60mmol,4.0 eq) were added and stirred at 100℃for 6hr. TLC (petroleum ether: ethyl acetate=.3:1) showed that after the reaction was completed, the reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was spun dry (water pump, 40 ℃) under reduced pressure to give the product ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } amino) methanoic acid-2-methylpropan-2-yl ester (47-2) (340 mg,1.06mmol, 49.30%) as a white solid, and the crude was directly taken to the next step.
The third step compound ({ 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } amino) methanoic acid-2-methylpropan-2-yl ester (47-2) (120 mg,0.376mmol,1.0 eq) was added to a reaction flask containing 4mol/L hydrochloric acid/1-4 dioxane solution (4 mL) and stirred for 4hr. TLC (petroleum ether: ethyl acetate=.3:1) monitored reaction was complete. The reaction mixture was dried under reduced pressure (water pump, 50 ℃ C.) to give crude 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-amine (47-3) as a white solid. LC-MS (ESI) [ m+h ] + =220.2.
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (99.8 mg,0.223mmol,1.0 eq)) was added to a reaction flask containing DMF (N, N-dimethylformamide) (5 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-diisopropylethylamine (0.074 mL,0.446mmol,2.0 eq) were stirred at room temperature (18 ℃) for 0.5hr, and then 1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-amine (47-3) (48.84 mg,0.223mmol,1 eq) was added and stirring was continued for 1hr. TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } propanamide (47-4) (60 mg,0.074mmol, 32.39%) as a white solid. LC-MS (ESI) [ m+h ] + = 650.5.
The fifth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } propionamide (47-4) (60 mg,0.074mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (0.74 mL,0.74 mmol,10 eq) was added and then warmed to 50℃and stirred for 16hr. TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [3- (pyridin-2-yl) propyl ] hexahydropyridin-4-yl } propionamide (47) (14.35 mg,0.0436mmol, 36.22%) as a white solid .1H NMR(400MHz,CDCl3)δ8.53(d,1H),7.61(d,1H),7.18–7.10(m,2H),5.44(s,1H),3.87(s,1H),3.23(d,1H),3.04(s,1H),2.84(s,1H),2.49–2.41(m,1H),2.32–2.20(m,3H),2.19–2.12(m,2H),2.04(d,2H),1.96(d,4H),1.89(s,1H),1.81–1.66(m,6H),1.64(s,3H),1.60–1.54(m,2H),1.41–1.36(m,1H),1.30(d,3H),1.08–1.01(m,2H),0.99(d,3H),0.95–0.91(m,1H),0.89–0.85(m,2H),0.81(d,6H).13C NMR(101MHz,CDCl3)δ172.25,161.64,149.28,136.32,131.86,128.69,122.79,121.08,79.06,77.34,77.22,77.02,76.70,57.94,54.32,53.98,52.30,46.18,38.85,37.76,36.96,36.53,36.14,35.46,33.62,32.12,28.06,27.66,26.86,26.83,24.95,21.92,21.85,19.66,15.37,14.31.LC-MS[M+H]+＝536.4.
Example 48
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [2- (methylamino) ethyl ] butanamide
The first step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (100 mg,0.216mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (98.59 mg, 0.59 eq, 1.2 eq) and N, N-diisopropylethylamine (N, N-diisopropylethylamine) (0.071 mL,0.432mmol,2.0 eq) was added, and stirring was continued at room temperature (10 ℃ C.) for 0.5hr, followed by [ (2-aminoethyl) (methyl) amino ] methanoic acid-2-methylpropan-2-yl ester (45.59 mg, 1.255 mmol, 1.9 eq). TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give crude product {9- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -6-oxyphenylene-2, 5-diazanon-2-yl } methanoic acid-2-methylpropan-2-yl ester (48-1) (80 mg,0.116mmol, 53.83%) as pale yellow solid, which was obtained by column chromatography (petroleum ether: 1:0to 2:1) .1H NMR(400MHz,CDCl3)δ6.37(s,1H),3.39(s,4H),3.17(dd,J＝11.2,4.5Hz,1H),2.88(s,3H),2.11(m,4H),1.91(m,3H),1.76–1.63(m,5H),1.58(s,3H),1.52(m,2H),1.46(s,9H),1.34(m,2H),0.96(m,3H),0.89(s,9H),0.87(s,3H),0.84(m,2H),0.80(s,3H),0.75(s,3H),0.02(d,2H).
The second step compound {9- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -6-oxy-2, 5-diazanon-2-yl } methano-2-methylpropan-2-yl ester (48-1) (80 mg,0.129mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (1.292 mL,1.292mmol,10 eq) a temperature was raised to 50℃and stirred for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a brown oil, which was extracted with ethyl acetate (20 mL), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure (water pump, 40 ℃) to give a crude product which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give {9- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -6-oxo-2, 5-diazanon-2-yl } methano-2-methylpropan-2-yl ester (48-2) (45 mg,0.080mmol, 62.20%) as a colorless solid ;1H NMR(400MHz,CDCl3)δ3.41(s,4H),3.22(dd,J＝11.5,4.6Hz,1H),2.88(s,3H),2.30–2.07(m,4H),1.93(dd,J＝29.9,15.9Hz,3H),1.71–1.60(m,6H),1.58(s,3H),1.54(s,1H),1.47(s,9H),1.35(dd,J＝15.9,12.5Hz,2H),1.09–1.00(m,2H),0.96(s,3H),0.94–0.83(m,3H),0.81(s,3H),0.79(s,3H).
The third step {9- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -6-oxo-2, 5-diazanon-2-yl } methanoic acid-2-methylpropan-2-yl ester (48-2) (45 mg,0.089mmol,1.0 eq) was added to a reaction flask containing methanol (0.1 mL), and after adding a solution of hydrogen chloride in methanol (0.223 mL,0.892mmol,10 eq), the mixture was stirred at room temperature (10 ℃) for 16hr. TLC (dichloromethane: methanol=5:1) monitored reaction was complete. Methanol (5 mL) is added into the reaction solution, the pH value is regulated to 8 by saturated sodium carbonate aqueous solution, then, the mixture is directly dried and stirred, and the product 4- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [2- (methylamino) ethyl ] butyramide (48) (29.82 mg,0.069mmol, 77.48%) is obtained as a white solid after separation and purification by column chromatography (dichloromethane: methanol=1:0 to 3:1) .1H NMR(400MHz,CDCl3)δ7.05(s,1H),3.58(d,J＝4.8Hz,2H),3.22(dd,J＝11.4,4.5Hz,1H),3.06–2.98(m,2H),2.64(s,3H),2.23(m,2H),2.18–1.99(m,3H),1.99–1.87(m,2H),1.80–1.70(m,2H),1.66(m,3H),1.59(s,3H),1.55(m,2H),1.43–1.28(m,2H),1.08(m,3H),0.98–0.92(s,3H),0.87(m,3H),0.81(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ174.35,132.56,127.54,79.09,77.32,77.21,77.00,76.69,54.32,53.98,50.30,38.83,37.91,36.97,36.81,36.53,36.39,34.10,33.55,32.12,28.67,28.08,27.66,24.39,21.95,21.85,19.60,15.40,14.30.LC-MS(ESI)[M+H]+＝405.25.
Example 49
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } propanamide
Reference example 47 conversion of 3- (pyridin-2-yl) propan-1-ol to 2- (pyridin-2-yl) ethan-1-ol gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {1- [2- (pyridin-2-yl) ethyl ] hexahydropyridin-4-yl } propanamide (49).1H NMR(400MHz,CDCl3)δ8.51(d,1H),7.62–7.58(m,1H),7.20(d,1H),7.13(m,1H),5.43(s,1H),3.86(s,1H),3.22(d,1H),3.10(s,3H),2.95(s,1H),2.49–2.30(m,4H),2.26–2.20(m,1H),2.17–2.10(m,2H),1.94(d,6H),1.80–1.72(m,2H),1.71–1.65(m,3H),1.63(s,3H),1.59(s,1H),1.41–1.24(m,4H),1.04(m,2H),0.97(d,3H),0.92–0.83(m,3H),0.80(d,6H).13C NMR(101MHz,CDCl3)δ172.25,160.27,149.30,136.37,131.88,128.69,123.18,121.24,79.06,77.34,77.23,77.03,76.71,58.33,54.33,53.98,52.28,46.28,38.85,37.76,36.96,36.53,35.90,35.47,33.63,32.30,32.13,28.06,27.66,24.96,21.92,21.85,19.66,15.37,14.31.LC-MS(ESI)[M+H]+＝522.5.
Example 50
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindol-5-yl) butanamide
Reference example 2 conversion of 1-methylpyrrolo [2,3-b ] pyridin-5-amine to 1-methylindol-5-amine gives the compound 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylindol-5-yl) butanamide (50).1H NMR(400MHz,CDCl3)δ7.80(s,1H),7.26(s,1H),7.12(s,1H),7.04(d,J＝2.9Hz,1H),6.43(d,J＝2.9Hz,1H),3.80(s,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.35(m,2H),2.18(m,2H),2.00(m,2H),1.88(m,2H),1.77(d,J＝13.2Hz,1H),1.69–1.65(m,3H),1.62(s,3H),1.58(m,2H),1.44–1.31(m,2H),1.05(m,3H),0.97(s,3H),0.93–0.83(m,3H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ171.11,134.23,132.55,130.21,129.63,128.54,127.67,115.90,112.77,109.25,101.04,79.12,77.33,77.21,77.01,76.69,54.33,54.00,38.83,37.87,37.59,36.99,36.54,33.60,32.91,32.14,28.56,28.06,27.67,24.55,21.94,21.88,19.67,15.36,14.30.LC-MS(ESI)[M+H]+＝477.3.
Example 51
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (6-phenylhexyl) propanamide
With reference to the example 46 of the present invention, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I), obtaining the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (6-phenylhexyl) propionamide (51).1H NMR(400MHz,CDCl3)δ7.32–7.26(m,2H),7.19-7.16(m,3H),5.51(s,1H),3.28–3.16(m,3H),2.63–2.55(m,2H),2.48–2.40(m,1H),2.37–2.21(m,2H),2.18–2.07(m,2H),2.05–1.87(m,3H),1.80-1.74(m,2H),1.73–1.65(m,3H),1.57(m,3H),1.52–1.44(m,2H),1.40–1.24(m,6H),1.17–0.98(m,3H),0.97(d,J＝6.1Hz,3H),0.94–0.83(m,3H),0.80(d,J＝8.0Hz,6H).13C NMR(101MHz,CDCl3)δ172.88,142.63,131.89,128.51,128.38,128.27,125.66,79.05,77.35,77.23,77.03,76.71,54.29,53.94,39.51,38.83,37.89,36.93,36.51,35.87,35.58,33.57,32.12,31.35,29.63,28.94,28.05,27.63,26.81,25.15,21.91,21.80,19.59,15.38,14.30.LC-MS(ESI):[M+H+]＝494.4.
Example 52
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-methoxyethyl) butanamide
Reference example 2 conversion of 1-methylpyrrolo [2,3-b ] pyridin-5-amine to 2-methoxyethyl-1-amine gives the compound 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-methoxyethyl) butanamide (52)1H NMR(400MHz,CDCl3)δ5.82(s,1H),3.46(s,4H),3.36(s,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.23–2.11(m,3H),2.06(dd,J＝17.8,9.5Hz,2H),1.93(m,3H),1.81–1.62(m,6H),1.59(s,3H),1.55(m,1H),1.42–1.26(m,2H),1.10–1.00(m,2H),0.97(s,3H),0.87(m,2H),0.80(d,J＝9.0Hz,6H).13C NMR(101MHz,CDCl3)δ173.06,132.53,127.55,79.09,77.35,77.24,77.04,76.72,71.33,58.77,54.30,53.96,39.12,38.83,37.82,36.97,36.56,36.52,33.56,32.09,28.54,28.06,27.65,24.46,21.93,21.86,19.59,15.39,14.29.LC-MS(ESI):[M+H+]＝406.2.
Example 53
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (8-phenyloctyl) propanamide
In the first step, (8-bromooctyl) benzene (0.5 g,1.86mmol,1.0 eq) and potassium phthalimide (0.41 g,2.23mmol,1.2 eq) were dissolved in DMF (N, N-dimethylformamide) (5 mL) and the reaction stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (50 mL. Times.2), and washed with water (60 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was purified by passing through a column with petroleum ether ethyl acetate=5:1 to give 2- (8-phenyloctyl) isoindole-1, 3-dione (53-1) (0.4 g, purity 95%, yield) 61.3％).1H NMR(400MHz,CDCl3)δ7.83(dt,J＝6.9,3.4Hz,2H),7.75–7.64(m,2H),7.26(t,J＝7.4Hz,2H),7.16(d,J＝6.3Hz,3H),3.67(t,J＝7.3Hz,1H),2.64–2.54(m,1H),1.62(ddd,J＝21.9,13.9,6.9Hz,2H),1.31(s,4H).
In the second step, 2- (8-phenyloctyl) isoindole-1, 3-dione (53-1) (0.15 g,0.45mmol,1.0 eq) was dissolved in ethanol (10 mL), hydrazine hydrate (0.11 mL,2.24mmol,5.0 eq) was added, the reaction was stirred at 80℃for 3h, and TLC (dichloromethane: methanol=10:1) showed completion of the reaction. After the completion of the reaction, the mixture was concentrated, extracted with chloroform (20 mL), washed with water (20 mL. Times.2), dried over anhydrous Na 2SO4, and the crude 8-phenyloct-1-amine (53-2) was directly fed to the next step. LC-MS (ESI) [ M+H + ] =206.2
In the third step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min, the reaction 8-phenyloct-1-amine (53-2) (45.8 mg,0.33mmol,1.0 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate=3:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1 column chromatography gave 3- [ (4 as,4br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (8-phenyloctyl) propanamide (53-3) (70 mg, purity 90%, yield 44.5%).
In the fourth step, 3- [ (4 aS,4bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (8-phenyloctyl) propionamide (53-3) (70 mg,0.11mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), and a tetrahydrofuran solution of TBAF (1.10 mL,1.10mmol,10.0 eq) was added and the reaction mixture was stirred overnight at 60 ℃. TLC (petroleum ether: ethyl acetate=2:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with petroleum ether ethyl acetate=3:1 to give 3- [ (4 as,4br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (8-phenyloctyl) propanamide (53) (16 mg, purity 94%, yield 24.6％).1H NMR(400MHz,CDCl3)δ7.29-7.25(m,2H),7.18-7.16(m,3H),5.50(s,1H),3.24-3.20(m,3H),2.64–2.56(m,2H),2.50–2.40(m,1H),2.39–2.21(m,2H),2.20-2.09(m,2H),2.05–1.86(m,3H),1.78-1.76(m,1H),1.75–1.67(m,3H),1.63(s,4H),1.63-1.59(m,2H),1.51–1.43(m,2H),1.37(dd,J＝13.4,3.6Hz,1H),1.35-1.25(m,7H),1.18–0.99(m,3H),0.97(s,3H),0.92(dd,J＝11.8,4.4Hz,1H),0.88-0.83(m,2H),0.81-0.79(m,6H).13C NMR(101MHz,CDCl3)δ172.87,142.83,131.90,128.50,128.38,128.23,125.58,79.04,77.33,77.22,77.02,76.70,54.28,53.93,39.54,38.82,37.87,36.93,36.50,35.95,35.55,33.57,32.11,31.47,29.67,29.38,29.22,28.04,27.62,26.91,25.13,21.90,21.79,19.58,15.37,14.29.LC-MS(ESI):[M+H+]＝522.50.
Example 54
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyrimidin-2-yl) heptyl ] propionamide
The first step of the compounds 2-bromopyrimidine (3 g,18.869mmol,1.0 eq) and hept-6-yn-1-ol (3.17 g,28.304mmol,1.5 eq) were added to a reaction flask containing acetonitrile (60 mL), and copper iodide (CuI) (0.04 g,0.189mmol,0.01 eq), bis (triphenylphosphine) palladium dichloride (0.29 g,0.377mmol,0.02 eq) and triethylamine (13.114 mL, 94.348 mmol,5.0 eq) were added under nitrogen and stirred for 2hr at 70 ℃. The reaction was monitored by TLC (dichloromethane: methanol=20:1). The reaction mixture was dried under reduced pressure (water pump, 45 ℃ C.) to give a crude product which was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 99:1) to give 7- (pyrimidin-2-yl) hept-6-yn-1-ol (54-1) (3 g,14.309mmol, 75.83%) as a yellowish brown oil .1H NMR(400MHz,CDCl3)δ8.71(d,J＝4.9Hz,2H),7.23(t,J＝4.9Hz,1H),3.68(t,J＝6.2Hz,2H),2.51(t,J＝7.0Hz,2H),1.76–1.67(m,2H),1.66–1.57(m,4H).
The second step, 7- (pyrimidin-2-yl) hept-6-yn-1-ol (54-1) (1 g,5.256mmol,1.0 eq) was added to a reaction flask containing methanol (10 mL), pd/C10% (0.5 g,0.470 mmol) was added and then the mixture was stirred for 2hr at 50℃under hydrogen atmosphere; LC-MS monitoring showed complete reaction, filtration of the reaction solution through celite, and spin-drying of the filtrate under reduced pressure (water pump, 50 ℃ C.) gave the product 7- (pyrimidin-2-yl) hept-1-ol (54-2) (1 g,4.632mmol, 88.13%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.67(d,J＝4.9Hz,2H),7.12(t,J＝4.9Hz,1H),3.66–3.62(m,2H),3.00–2.92(m,2H),1.87–1.80(m,2H),1.59–1.54(m,2H),1.38(s,8H).
In a third step, 7- (pyrimidin-2-yl) heptan-1-ol (54-2) (1 g,5.147mmol,1.0 eq) was added to a reaction flask containing acetonitrile (20 mL), triphenylphosphine (2.43 g,9.265mmol,1.8 eq) and carbon tetrabromide (3.07 g,9.265mmol,1.8 eq) were added at 0deg.C, and stirring was continued at room temperature (18 ℃) for 1hr, and the reaction was monitored by TLC (petroleum ether: ethyl acetate=1:1). The reaction solution was concentrated under reduced pressure (water pump, 45 ℃) and water (20 mL) was added, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give the crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0to5:1) to give the product 2- (7-bromoheptyl) pyrimidine (54-3) (220 mg,0.809mmol, 20.84%) as a colorless oil .1H NMR(400MHz,CDCl3)δ8.69(d,J＝4.9Hz,2H),7.16(t,J＝4.9Hz,1H),3.40(t,J＝6.9Hz,2H),3.02–2.94(m,2H),1.91–1.79(m,4H),1.48–1.35(m,6H).
The fourth step compound 2- (7-Bromoheptyl) pyrimidine (54-3) (700 mg, 2.720 mmol,1.0 eq) and potassium phthalimide (608.20 mg,3.266mmol,1.2 eq) were added to a reaction flask containing DMF (N, N-dimethylformamide) (10 mL) and stirred at room temperature (18 ℃ C.) for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction was substantially complete. The reaction mixture was taken up in water (40 mL), extracted with ethyl acetate (30 mL. Times.3), the organic phases were combined, washed with water (30 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give the crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1) to give the product 2- [7- (pyrimidin-2-yl) heptyl ] isoindole-1, 3-dione (54-4) (600 mg,1.855mmol, 68.16%) as a pale yellow oil .1H NMR(400MHz,CDCl3)δ8.68(d,J＝4.9Hz,2H),7.87–7.81(m,2H),7.74–7.67(m,2H),7.14(t,J＝4.9Hz,1H),3.67(t,J＝7.3Hz,2H),3.00–2.93(m,2H),1.83(dd,J＝14.6,7.2Hz,2H),1.72–1.61(m,2H),1.35(d,J＝19.1Hz,6H).
The fifth step compound 2- [7- (pyrimidin-2-yl) heptyl ] isoindole-1, 3-dione (54-4) (150 mg, 0.460 mmol,1.0 eq) was added to a reaction flask containing EtOH (2 mL), hydrazine hydrate (0.056 mL,0.928mmol,2.0 eq) was added and the mixture was heated to 80℃and stirred for 16hr. LC-MS monitoring of the reaction showed the starting material was essentially complete. The reaction mixture was directly stirred and purified by column chromatography (dichloromethane: methanol=1:0 to 1:1) to give 7- (pyrimidin-2-yl) hept-1-amine (54-5) (60 mg,0.279mmol, 60.23%) as a pale yellow oil .1H NMR(400MHz,MeOD)δ8.70(d,J＝5.0Hz,2H),7.32(s,1H),2.97–2.88(m,2H),2.70(t,J＝7.3Hz,2H),1.81(m,2H),1.51(m,2H),1.38(s,6H).
The sixth step compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (I) (80 mg,0.239mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (91.51 mg,0.241mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.066 mL,0.401mmol,2.0 eq) was added and stirring was continued for 1hr after stirring at room temperature (18 ℃) for 0.5hr, followed by addition of 7- (pyrimidin-2-yl) hept-1-amine (54-5) (60 mg,0.310mmol,1.3 eq). TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a pale yellow crude product, which was purified by prrp-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyrimidin-2-yl) heptyl ] propionamide (54-6) (80 mg,0.115mmol, 57.40%) as an off-white solid ;1H NMR(400MHz,CDCl3)δ8.81(d,J＝4.9Hz,2H),7.35(s,1H),5.52(s,1H),3.28–3.15(m,3H),3.11(t,J＝7.7Hz,2H),2.37(m,2H),2.25(m,1H),2.15(m,2H),2.08–1.84(m,6H),1.70(d,J＝13.4Hz,3H),1.63(s,3H),1.60–1.45(m,4H),1.44–1.27(m,9H),1.12–0.94(m,3H),0.93–0.86(s,11H),0.82(s,3H),0.76(s,3H),0.04(d,J＝2.6Hz,6H).
The seventh step 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyrimidin-2-yl) heptyl ] propanamide (54-6) (80 mg,0.128mmol,1.0 eq) was added to a reaction flask containing THF (1 mL), tetrabutylammonium fluoride in tetrahydrofuran (1.282 mL,1.282mmol,10 eq) was added and then warmed to 50deg.C and stirred for 16hr; TLC (dichloromethane: methanol=10:1) monitored the reaction for completion. The reaction solution was dried under reduced pressure (water pump, 40 ℃) and water (20 mL) was added, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a pale yellow crude product, which was isolated and purified by prrp-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyrimidin-2-yl) heptyl ] propionamide (54) as a white solid .1H NMR(400MHz,CDCl3)δ8.68(d,J＝4.9Hz,2H),7.14(t,J＝4.9Hz,1H),5.43(s,1H),3.22(dd,J＝11.5,4.8Hz,3H),3.00–2.93(m,2H),2.56–2.38(m,1H),2.37–2.28(m,1H),2.27–2.07(m,3H),2.05–1.93(m,2H),1.92–1.82(m,3H),1.82–1.74(m,2H),1.73–1.65(m,3H),1.62(s,3H),1.50–1.44(m,2H),1.35(m,7H),1.08–1.00(m,2H),0.97(d,J＝5.8Hz,3H),0.94–0.83(m,3H),0.80(d,J＝7.9Hz,6H).13C NMR(101MHz,CDCl3)δ172.81,171.47,156.98,131.92,128.47,118.43,79.05,77.34,77.22,77.02,76.70,54.29,53.94,39.52,39.41,38.83,37.88,36.93,36.50,35.59,33.57,32.11,29.63,29.21,29.09,28.55,28.05,27.63,26.81,25.14,21.90,21.79,19.58,15.37,14.29.LC-MS(ESI):[M+H+]＝510.4.
Example 55
Preparation of the Compound N- (1-cyclopropylin-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
The first step compound 5-nitro-1H-indole (100 mg, 0.611 mmol,1.0 eq), cyclopropylborodiol (105.95 mg,1.233mmol,2.0 eq) and sodium carbonate (130.73 mg,1.233mmol,2.0 eq) were dissolved in 1, 2-dichloroethane (2 mL) and added to a hot solution of bipyridine (96.32 mg, 0.611 mmol,1.0 eq) and copper acetate (112.01 mg, 0.611 mmol,1.0 eq) in 1, 2-dichloroethane (2 mL) and stirred under an oxygen atmosphere at 70℃for 4hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was added with dilute hydrochloric acid (1M) aqueous solution (10 mL) and saturated ammonium chloride (20 mL) aqueous solution, extracted with dichloromethane (20 mL x 3), the organic phases were combined, dried by spin-drying under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-19:1) to give the product 1-cyclopropyl-5-nitroindole (55-1) (90 mg,0.401mmol, 64.95%) as a white solid .1H NMR(400MHz,CDCl3)δ8.56(d,J＝2.0Hz,1H),8.13(dd,J＝9.0,2.1Hz,1H),7.58(d,J＝9.0Hz,1H),7.27(d,J＝3.3Hz,1H),6.61(d,J＝3.2Hz,1H),3.45–3.37(m,1H),1.20–1.13(m,2H),1.09–1.01(m,2H).
In the second step, 1-cyclopropyl-5-nitroindole (55-1) (90 mg,0.445mmol,1.0 eq) was added to a reaction flask containing ethanol (3 mL) and water (1 mL), and after adding iron filings (173.97 mg,3.116mmol,7.0 eq) and ammonium chloride (285.69 mg, 5.3411 mmol,12 eq), the temperature was raised to 80℃and stirred for 4h. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was filtered, ethyl acetate (30 mL) was added, washed with water (20 mL x 2) and dried under reduced pressure (water pump, 45 ℃) to give crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-4:1) to give 1-cyclopropylindol-5-amine (55-2) (30 mg,0.157mmol, 35.22%) as a brown solid .1H NMR(400MHz,CDCl3)δ7.38(d,J＝8.6Hz,1H),7.05(d,J＝3.1Hz,1H),6.98(d,J＝1.8Hz,1H),6.74(dd,J＝8.5,1.9Hz,1H),6.25(d,J＝3.1Hz,1H),3.32–3.23(m,1H),1.03–0.96(m,4H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (60 mg,0.130mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), HATU (54.23 mg,0.143mmol,1.1 eq) and diisopropylethylamine (0.043 mL, 0.255 mmol,2.0 eq) were added, followed by stirring at room temperature (25 ℃) for 0.5hr, and then 1-cyclopropylindol-5-amine (55-2) (26.80 mg,0.156mmol,1.2 eq) was added, followed by stirring for 1hr. TLC (petroleum ether: ethyl acetate=3:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, and after washing with water (20 mL x 3), the organic phase was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-9:1) to give the product N- (1-cyclopropylin-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydro phenanthryl ] butanamide (55-3) (73 mg,0.106mmol, 82.14%) as a pale yellow solid .1H NMR(400MHz,CDCl3)δ7.78(d,J＝1.7Hz,1H),7.49(d,J＝8.7Hz,1H),7.24(d,J＝1.9Hz,1H),7.12(d,J＝3.1Hz,1H),6.38(d,J＝3.0Hz,1H),3.33(ddd,J＝10.4,6.8,3.7Hz,1H),3.18(dd,J＝11.1,4.5Hz,1H),2.43–2.29(m,2H),2.25–2.10(m,3H),2.05–1.78(m,5H),1.73–1.67(m,4H),1.63(s,3H),1.55(dd,J＝14.7,10.5Hz,3H),1.38(s,1H),1.06(dd,J＝12.9,6.1Hz,3H),0.94(s,2H),0.89(s,10H),0.88(s,3H),0.85(s,1H),0.82(s,3H),0.80(s,1H),0.75(s,3H),0.04(d,J＝2.7Hz,6H).
The fourth step compound N- (1-cyclopropylin-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (55-3) (70 mg,0.113mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.135 mL,1.135mmol,10 eq) was added thereto, followed by heating to 50℃and stirring for 16hr. TLC (petroleum ether: ethyl acetate=3:1) monitored reaction completion. The reaction solution was dried under reduced pressure (water pump, 45 ℃) to give a brown oil, ethyl acetate (30 mL) was added, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by prep-TLC (petroleum ether: ethyl acetate=3:1) to give the product N- (1-cyclopropylin-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butyramide (55) (42.56 mg,0.073mmol, 64.68%) as a white solid .1H NMR(400MHz,MeOD)δ7.74(d,J＝1.7Hz,1H),7.48(d,J＝8.7Hz,1H),7.24(dd,J＝8.7,1.8Hz,1H),7.18(d,J＝3.1Hz,1H),6.33(d,J＝3.1Hz,1H),3.38–3.33(m,1H),3.15(m,1H),2.40–2.30(m,2H),2.28–2.11(m,3H),2.08–1.96(m,2H),1.87(m,2H),1.77(m,1H),1.74–1.64(m,4H),1.62(s,3H),1.61–1.58(m,1H),1.41(m,2H),1.14–1.02(m,4H),0.96(m,1H),0.94(s,3H),0.90(m,2H),0.85(s,3H),0.83(m,1H),0.77(s,3H).13C NMR(101MHz,MeOD)δ173.11,135.00,132.48,130.73,128.80,128.32,127.16,115.80,112.75,109.64,100.65,78.38,54.46,54.24,38.52,37.82,36.92,36.37,36.23,33.26,32.10,28.04,27.26,26.94,26.46,24.49,21.71,21.62,18.42,14.73,13.40,5.25.LC-MS(ESI)[M+1]+＝503.3.
Example 56
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyrazin-2-yl) heptyl ] propionamide
Reference example 54 conversion of 2-bromopyrimidine to 2-bromopyrazine gave the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyrazin-2-yl) heptyl ] propionamide (56).1H NMR(400MHz,CDCl3)δ8.51–8.41(m,3H),5.43(s,1H),3.20(dd,J＝11.5,4.6Hz,3H),2.85–2.79(m,2H),2.46–2.36(m,1H),2.32(m,1H),2.26–2.18(m,1H),2.16–2.06(m,2H),2.00–1.83(m,4H),1.74(m,3H),1.65(dd,J＝8.1,4.3Hz,2H),1.60(s,3H),1.57(m,1H),1.48–1.41(m,2H),1.33(dd,J＝15.4,10.0Hz,8H),1.01(m,3H),0.95–0.89(s 3H),0.88–0.81(m,2H),0.78(d,J＝7.8Hz,6H).13C NMR(101MHz,CDCl3)δ172.83,157.85,144.55,143.96,142.07,131.93,128.45,79.03,77.34,77.22,77.02,76.70,54.31,53.96,39.47,38.83,37.91,36.95,36.51,35.61,35.40,33.57,32.12,29.65,29.26,29.12,29.04,28.05,27.64,26.78,25.14,21.91,21.80,19.56,15.37,14.28.LC-MS[M+H]+＝510.3.
Example 57
Preparation of the Compound 3- [ ({ 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) methyl ] benzene-1-carboxamide
The first-step compound 3- (aminomethyl) benzene-1-carbonitrile (200 mg, 1.313 mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (5 mL), and di-tert-butyl dicarbonate (0.417 mL,1.816mmol,1.2 eq) and triethylamine (0.426 mL,3.026mmol,2.0 eq) were added and stirred at room temperature (18 ℃ C.) for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. After methylene chloride (20 mL) was added to the reaction solution, the mixture was washed with water (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1) to give a product { [ (3-cyanophenyl) methyl ] amino } methanoic acid-2-methylpropan-2-yl ester (57-1) (350 mg,1.480mmol, 97.77%) as a colorless oil .1H NMR(400MHz,CDCl3)δ7.55(dd,J＝14.5,9.1Hz,3H),7.44(t,J＝7.7Hz,1H),4.95(s,1H),4.35(s,2H),1.47(s,9H).
The second step compound { [ (3-cyanophenyl) methyl ] amino } methanoic acid-2-methylpropan-2-yl ester (57-1) (150 mg, 0.640 mmol,1.0 eq) was dissolved in ethanol (1 mL) and added to a reaction flask containing water (1 mL) of sodium hydroxide (51.66 mg,1.292mmol,2.0 eq), hydrogen peroxide (0.100 mL,1.292mmol,2.0 eq) was slowly added dropwise and stirred at room temperature (18 ℃ C.) for 2hr. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction mixture was quenched with saturated aqueous sodium sulfite (10 mL), extracted with dichloromethane (20 mL), and the organic phase was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=1:0 to 24:1) to give { [ (3-formamidophenyl) methyl ] amino } methanoic acid-2-methylpropan-2-yl ester (57-2) (140 mg,0.559mmol, 86.61%) as a white solid .1H NMR(400MHz,CDCl3)δ7.75(s,1H),7.70(d,J＝7.4Hz,1H),7.44(dt,J＝15.0,7.5Hz,2H),6.16(s,1H),5.69(s,1H),4.95(s,1H),4.37(d,J＝5.5Hz,2H),1.46(s,9H).
The third step compound { [ (3-carboxamidophenyl) methyl ] amino } methanoic acid-2-methylpropan-2-yl ester (57-2) (140 mg,0.559mmol,1.0 eq) was added to a reaction flask containing methanol (0.5 mL), and after adding a methanol solution of hydrogen chloride (1.390 mL,5.593mmol,10 eq), stirred at room temperature (18 ℃ C.) for 1hr. LC-MS monitored reaction was complete. The reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) and replaced with oil pump to give the product 3- (aminomethyl) benzene-1-carboxamide (57-3) (80 mg,0.53 mmol, 95.24%) as a white solid. LC-MS (ESI) [ m+1] + =151.1 (ms+1).
The fourth step compound 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (81.34 mg,0.214mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.059 mL, 0.317 mmol,2.0 eq) were added and stirring was continued for 1hr after stirring at room temperature (18 ℃) for 0.5hr, and 3- (aminomethyl) benzene-1-carboxamide (40.16 mg,0.26 mmol,1.5 eq) was added. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=1:0 to 24:1) to give 3- [ ({ 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) methyl ] benzene-1-carboxamide (57-4) (70 mg,0.108mmol, 60.83%) as a white solid .1H NMR(400MHz,CDCl3)δ7.76(s,1H),7.70(d,J＝7.4Hz,1H),7.45(d,J＝5.3Hz,2H),6.14(s,1H),5.85(s,1H),5.70–5.54(m,1H),4.48(s,2H),3.17(dd,J＝11.0,4.4Hz,1H),2.39(m,3H),2.22(m,2H),1.95(m,5H),1.69(m,3H),1.59(s,3H),1.33(m,2H),0.97(m,2H),0.88(d,J＝6.7Hz,14H),0.78(s,3H),0.74(s,3H),0.03(d,J＝2.5Hz,6H).
The fifth step compound 3- [ ({ 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) methyl ] benzene-1-carboxamide (57-4) (70 mg,0.120mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.205 mL,1.205mmol,10 eq) was added thereto, followed by heating to 50℃and stirring for 16hr. TLC (dichloromethane: methanol=10:1) monitored the reaction was essentially complete. The reaction solution was concentrated under reduced pressure (water pump, 40 ℃) to give a pale yellow oil, which was extracted with water (20 mL), the organic phases were combined, washed with water (30 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give crude product 3- [ ({ 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] propionyl } amino) methyl ] benzene-1-carboxamide (57) (14.44 mg,0.030mmol, 24.65%) as a white solid by separating and purifying the crude product by prep-TLC (dichloromethane: methanol=10:1) .1H NMR(400MHz,CDCl3)δ7.69(s,1H),7.63(d,J＝6.2Hz,1H),7.36(d,J＝11.9Hz,2H),6.12(s,1H),5.81(s,1H),5.63(s,1H),4.41(s,2H),3.14(d,J＝7.7Hz,1H),2.38(m,1H),2.27(m,2H),2.10(m,2H),1.78(m,4H),1.59(m,4H),1.52(s,3H),1.29(m,2H),0.99(m,2H),0.89(s,3H),0.77(m,2H),0.72(s,6H).13C NMR(101MHz,CDCl3)δ171.93,167.92,138.21,132.74,130.61,130.47,128.01,127.71,125.92,125.30,78.02,76.31,76.20,75.99,75.68,53.24,52.87,42.26,37.79,36.83,35.88,35.46,34.40,32.49,31.08,27.01,26.59,24.04,20.86,20.69,18.54,14.34,13.24.LC-MS[M+H]+＝467.3.
Example 58
Preparation of the Compound N- (hexahydropyridin-4-ylmethyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide hydrochloride
The first step compound 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (122.01 mg,0.321mmol,1.2 eq) and N, N-Diisopropylethylamine (DIEA) (0.088 mL, 0.545 mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by the addition of 4- (aminomethyl) hexahydropyridine-1-carboxylic acid-2-methylpropan-yl ester (85.96 mg,0.401mmol,1.5 eq). TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction showed completion. The reaction solution was added with water (25 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3), and dried under reduced pressure to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1), to give the product 4- [ ({ 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) methyl ] piperidine-1-carboxylic acid-2-methylpropan-2-yl ester (58-1) (90 mg,0.126mmol, 46.96%) as a colorless oil .1H NMR(400MHz,CDCl3)δ5.55(s,1H),4.10(m,2H),3.16(m,3H),2.67(t,2H),2.44(m,1H),2.27(m,2H),2.15(m,2H),1.97(m,4H),1.66(m,8H),1.63(s,3H),1.45(s,9H),1.11(m,3H),1.04–0.95(m,2H),0.88(s,12H),0.83(m,2H),0.80(s,3H),0.75(s,3H),0.03(d,J＝3.0Hz,6H).
The second step compound, 2-methylpropan-2-yl 4- [ ({ 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) methyl ] hexahydropyridine-1-carboxylate (58-1) (90 mg,0.140mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (1.3995 mL, 1.3995 mmol,10 eq) was warmed to 50deg.C and stirred for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil. Water (15 mL) was added, the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, the filtrate was dried under reduced pressure (water pump, 40 ℃) and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product 4- [ ({ 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) methyl ] piperidine-1-carboxylic acid-2-methylpropan-2-yl ester (58-2) (40 mg,0.068mmol, 48.61%) as a white solid .1H NMR(400MHz,CDCl3)δ5.52(s,1H),4.10(m,2H),3.22(dd,J＝11.5,4.5Hz,1H),3.14(d,J＝4.5Hz,2H),2.67(t,J＝12.2Hz,2H),2.30(m,3H),2.19–2.10(m,2H),1.96(m,3H),1.75(m,2H),1.69(s,5H),1.63(s,3H),1.45(s,9H),1.17–1.00(m,5H),0.97(s,3H),0.88(t,J＝11.1Hz,4H),0.80(d,J＝8.3Hz,6H).
The third step compound 4- [ ({ 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) methyl ] hexahydropyridine-1-carboxylic acid 2-methylpropan-2-yl ester (58-2) (40 mg,0.075mmol,1.0 eq) was added to a reaction flask containing anhydrous methanol (1 mL), and after adding a hydrogen chloride/methanol solution (0.188 mL,0.754mmol,10 eq) was stirred at room temperature (18 ℃) for 1hr. TLC (dichloromethane: methanol=5:1) monitored the reaction was complete. Saturated aqueous sodium bicarbonate solution was added dropwise to the reaction mixture to adjust the pH to 7-8, followed by spin drying under reduced pressure (water pump, 50 ℃) to give a crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 3:1) to give the product N- (hexahydropyridin-4-ylmethyl) -3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide hydrochloride (58) (22.1 mg,0.049mmol, 64.69%) as a white solid .1H NMR(400MHz,DMSO)δ8.83(s,1H),8.53(s,1H),7.90(s,1H),4.34(d,J＝5.0Hz,1H),3.22(d,J＝12.5Hz,2H),3.04–2.93(m,2H),2.89(dd,J＝13.2,6.5Hz,1H),2.80(d,J＝11.5Hz,2H),2.35–2.26(m,1H),2.13(m,3H),1.94(m,3H),1.63(m,8H),1.56(s,3H),1.50(m,2H),1.36–1.27(m,2H),1.08–0.91(m,3H),0.88(s,3H),0.79(m,2H),0.76(s,3H),0.69(s,3H).
13C NMR(101MHz,CDCl3)δ173.44,131.64,128.62,79.03,77.35,77.23,77.03,76.71,54.26,53.88,44.25,43.76,38.83,37.88,36.91,36.50,35.45,34.10,33.56,32.12,29.63,29.34,28.07,27.62,26.45,25.10,21.90,21.80,19.63,15.41,14.31.LC-MS[M+H]+＝431.2.
Example 59
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- { [ 3-cyano-1- (pyrazin-2-ylmethyl) indol-5-yl ] amino } methanamide
In a first step, under nitrogen, the reaction pyrazin-2-yl-methanol (200 mg,1.816 mmol), triethylamine (0.505 mL,3.632 mmol) and solvent dichloromethane (6 mL) were added to a 50mL three-necked flask, cooled to 0℃and methylsulfonyl chloride (416.01 mg,3.632 mmol) was added and the reaction stirred at 0deg.C for 3hr. LC-MS monitored the reaction. After completion of the reaction, the reaction solution was diluted with methylene chloride. Washed with water and saturated brine. The dichloromethane layer was dried over anhydrous sodium sulfate, filtered and concentrated to give the product pyrazin-2-ylmethyl methanesulfonate (59-1) (150 mg, 0.719 mmol, 39.50%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =189.1.
In a second step, 5-nitro-1H-indole-3-carbonitrile (59-1) (100 mg, 0.284 mmol) and solvent N, N-dimethylformamide (6 mL) were added to a 50mL three-necked flask, the temperature was reduced to 0℃and sodium hydride (29.62 mg, 0.8011 mmol) was added to the flask and stirred for 0.5hr, and then pyrazin-2-ylmethyl mesylate ((120 mg,0.641 mmol) was obtained as a product, the reaction mixture was stirred at room temperature under N2 for 3hr, LC-MS monitored for reaction, water (15 mL) was added, extracted with ethyl acetate (10 mL. Times.3), the organic phases were combined, washed with saturated brine (10 mL. Times.2) and then dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0:5:1) to give 5-nitro-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile (59-2) (120 mg, 0.280.38 mmol; colorless oil [ LC-387M ] 25. +%).
In a third step, the reaction 5-nitro-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile (59-2) (120 mg,0.430 mmol), palladium on carbon 10% (20 mg,0.188 mmol) and solvent methanol (8 mL) were added to a 50mL three-necked flask, and the reaction was stirred at room temperature under H2 for 0.5H. After the reaction was completed, the reaction solution was filtered by suction, and the organic phase was dried by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) and separated and purified to give the product 5-amino-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile (59-3) (70 mg, 0.255 mmol, 58.81%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =250.1.
The fourth step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (70 mg,0.209mmol,1.0 eq) was added to a reaction flask containing toluene (2 mL), diphenyl azide phosphate (57.59 mg,0.209mmol,1.05 eq) and triethylamine (0.029 mL,0.209mmol,2.0 eq) were added and stirred at room temperature (90 ℃) for 1hr, the reaction mixture was cooled to room temperature, tetrahydrofuran (2 mL) was added, and stirring was continued for 18hr with 5-amino-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile (59-3) (52.17 mg,0.209mmol,1.0 eq). LC-MS monitored reaction was complete. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } -1- { [ 3-cyano-1- (pyrazin-2-ylmethyl) indol-5-yl ] amino } methanamide (59) (330 mg,0.045mmol, 20.39%) as a white solid .1H NMR(400MHz,CDCl3)δ78.54(m,3H),8.10(s,1H),7.75(d,J＝1.8Hz,1H),7.45(d,J＝8.9Hz,1H),7.23(s,1H),5.61(s,2H),3.25(m,1H),3.12(m,2H),2.29(m,3H),2.01(m,2H),1.89(m,1H),1.74(m,2H),1.66(s,1H),1.63(s,3H),1.34(d,J＝7.3Hz,2H),1.28(s,3H),1.05(ddd,J＝15.8,10.3,6.3Hz,2H),0.95(d,J＝5.5Hz,3H),0.90(s,1H),0.84(d,J＝7.5Hz,3H),0.78(d,J＝8.9Hz,3H).13C NMR(101MHz,CDCl3)δ157.30,151.79,144.51,143.89,142.98,136.79,134.84,132.00,130.07,129.08,128.33,117.76,115.27,111.16,109.40,84.98,78.37,54.42,54.19,49.43,38.52,37.88,36.89,36.21,33.25,32.16,29.37,29.06,27.24,26.93,21.68,21.56,18.50,14.71,13.40.LC-MS(ESI)[M+H]+＝581.30.
Example 60
Preparation of Compound 3- ({ 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) propionic acid
In the first step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (60 mg,0.18mmol,1.0 eq), HATU (80.8 mg,0.21mmol,1.2 eq) and N, N-diisopropylethylamine (30.1 mg,0.23mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3 mL), and after stirring for 2min, methyl 3-aminopropionate (32.5 mg,0.23mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (15 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing it through a column with petroleum ether ethyl acetate=10:1 to give methyl 3- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) propionate (60-1) (40 mg, purity 90%, yield 47.8%).
In the second step, methyl 3- ({ 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) propionate (60-1) (40 mg,0.09mmol,1.0 eq) was dissolved in methyl (2 mL), and a 20NaOH% aqueous solution (0.2 mL) was added under ice bath, and the reaction solution was stirred at room temperature for 2h. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was concentrated and extracted with ethyl acetate (20 mL. Times.2), followed by washing with water (20 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and the crude was purified by column chromatography with dichloromethane: methanol=20:1-5:1 to give 3- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) propanoic acid (60) (3.53 mg, 85% purity, yield) as a white solid after concentration 28.9％).1H NMR(400MHz,DMSO)δ7.84(s,1H),4.32(s,1H),3.19(dq,J＝13.0,6.6Hz,2H),3.05–2.95(m,1H),2.35–2.21(m,2H),2.18–2.06(m,2H),1.99–1.88(m,2H),1.84(d,J＝15.3Hz,1H),1.74–1.58(m,3H),1.55(s,3H),1.52–1.43(m,2H),1.39–1.15(m,5H),1.11–0.81(m,6H),0.81–0.64(m,7H).13C NMR(101MHz,DMSO)δ172.29,132.53,127.19,77.40,54.40,54.00,38.93,37.96,37.06,36.55,35.40,35.12,34.85,33.52,32.16,29.48,28.65,27.91,25.52,22.05,21.87,19.73,16.33,14.57.LC-MS(ESI)[M+H]+＝406.2.
Example 61
Preparation of the Compound N- [6- (4-fluorophenyl) hexyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
Reference example 51 conversion of benzaldehyde to p-fluorobenzaldehyde gave the compound N- [6- (4-fluorophenyl) hexyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] propanamide (61)1H NMR(400MHz,CDCl3)δ7.11(dd,J＝8.3,5.6Hz,2H),6.95(t,J＝8.7Hz,2H),5.46(s,1H),3.24-3.20(m,3H),2.57(t,J＝7.7Hz,2H),2.47–2.06(m,5H),2.04–1.84(m,3H),1.80–1.56(m,11H),1.47-1.44(m,2H),1.39–1.29(m,5H),1.08–0.92(m,5H),0.85-0.79(m,8H).13C NMR(101MHz,CDCl3)δ172.84,162.38,159.97,138.14,131.91,129.66,129.58,128.43,115.05,114.84,79.04,54.31,53.96,39.38,38.83,37.91,36.95,36.51,35.61,35.01,33.57,32.13,31.44,29.65,28.81,28.05,27.64,26.77,25.14,21.91,21.80,19.56,15.36,14.29.LC-MS(ESI)[M+H]+＝512.4.
Example 62
Preparation of the Compound N- (2-aminoethyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionamide
In the first step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (120 mg,0.27mmol,1.0 eq), HATU (122.0 mg,0.32mmol,1.2 eq) and N, N-diisopropylethylamine (44.9 mg,0.35mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min the reaction [ (2-aminoethyl) amino ] methanoic acid-2-methylpropan-2-yl ester (55.7 mg,0.35mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate=3:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 3:1 column passed to give { [2- ({ 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) ethyl ] amino } methanoic acid-2-methylpropan-2-yl ester (62-1) (80 mg, 90% purity, 45.6% yield) the product was directly taken to the next step.
In the second step, 2-methylpropan-2-yl [2- ({ 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) ethyl ] amino } methanoate (62-1) (80 mg,0.15mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), and a solution of TBAF (1.52 mL,1.52mmol,10.0 eq) in tetrahydrofuran was added and the reaction mixture was stirred overnight at 60 ℃. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing methanol=20:1 through a column, and concentrated to give { [2- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) ethyl ] amino } methanoic acid-2-methylpropan-2-yl ester (62-2) (40 mg, purity 90%, yield 49.6%).
In the third step, 2-methylpropan-2-yl { [2- ({ 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) ethyl ] amino } methanoate (62-2) (40 mg,0.08mmol,1.0 eq) was dissolved in a hydrogen chloride-methanol solution (4 mol/L,2 mL), and the reaction solution was stirred at room temperature for 2hrs. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The crude product was concentrated by passing through a column with methylene chloride, methanol=20:1-10:1, and concentrated to give white solid N- (2-aminoethyl) -3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (62) (4.0 mg, purity 91.0%, yield) 11.5％).1H NMR(400MHz,DMSO)δ7.95(t,J＝5.5Hz,1H),3.12(dd,J＝12.7,6.4Hz,2H),3.02–2.96(m,1H),2.66(t,J＝6.4Hz,2H),2.32–2.23(m,1H),2.21–2.05(m,3H),2.03–1.78(m,4H),1.68–1.58(m,3H),1.58–1.45(m,5H),1.39–1.27(m,1H),1.23(s,1H),1.07–0.92(m,3H),0.90(d,J＝8.6Hz,3H),0.80–0.72(m,5H),0.71-0.69(m,3H).13C NMR(101MHz,DMSO)δ172.73,132.67,127.24,77.41,54.44,54.04,49.05,40.57,38.99,38.01,36.93,36.54,35.16,33.51,32.17,28.65,27.90,25.45,22.14,21.92,19.76,16.28,14.65.LC-MS(ESI)[M+H]+＝377.2.
Example 63
Preparation of the Compound N-decyl-N-ethyl-3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
The first step compound decan-1-amine (1000 mg, 6.317 mmol,1.0 eq) was added to a reaction flask containing dichloromethane (10 mL), followed by addition of di-tert-butyl dicarbonate (1.4631 mL, 6.317 mmol,1.0 eq) and triethylamine (1.767 mL, 12.015 mmol,2.0 eq) and stirring at room temperature (20deg.C) for 16hr. TLC (petroleum ether: ethyl acetate=5:1) showed dot formation with reduced polarity; dichloromethane (20 mL) was added to the reaction solution, which was washed with water (20 mL. Times.2), dried under reduced pressure (water pump, 40 ℃ C.) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1), and the product (decylamino) methanoic acid-2-methylpropan-2-yl ester (63-1) (1500 mg,5.827mmol, 91.66%) was collected as a white solid .1H NMR(400MHz,CDCl3)δ3.10(t,J＝7.1Hz,2H),1.50–1.41(m,11H),1.37–1.17(m,15H),0.88(t,J＝6.9Hz,3H).
The second step compound (decylamino) 2-methylpropan-2-yl ester (63-1) (200 mg,0.777mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), sodium hydrogen (40.40 mg,1.010mmol,1.3 eq) was added at 0deg.C, followed by stirring for 0.5hr, and bromoethane (0.075 mL,1.010mmol,1.3 eq) was added, followed by returning to room temperature (18 ℃ C.) and stirring for 16hr. TLC (petroleum ether: ethyl acetate=5:1) monitored the reaction was complete. The reaction mixture was quenched by dropwise addition of saturated aqueous ammonium chloride (5 mL), water (10 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give a crude product which was isolated and purified by column chromatography (petroleum ether) to give [ decyl (ethyl) amino ] methanoic acid-2-methylpropan-2-yl ester (63-2) (200 mg,0.701mmol, 90.18%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.20(t,J＝7.1Hz,2H),3.17–3.10(m,2H),1.53–1.47(m,2H),1.45(s,8H),1.26(s,14H),1.09(t,J＝7.1Hz,3H),0.88(t,J＝6.8Hz,4H).
The third step compound [ decyl (ethyl) amino ] methanoic acid-2-methylpropan-2-yl ester (63-2) (100 mg,0.350mmol,1.0 eq) was added to a reaction flask containing methanol (1 mL), and after adding a methanol solution of hydrogen chloride (0.876 mL,3.503mmol,10 eq), it was stirred at room temperature (18 ℃) for 16hr. TLC (petroleum ether: ethyl acetate=5:1) monitored the starting material reaction. The reaction solution was dried under reduced pressure (water pump, 50 ℃ C.) and was further dried by oil pump to give decyl (ethyl) amine (63-3) (60 mg,0.324mmol, 92.41%) as a white solid .1H NMR(400MHz,MeOD)δ3.04(q,J＝7.3Hz,2H),3.00–2.93(m,2H),1.72–1.62(m,2H),1.32(dd,J＝22.0,14.7Hz,16H),0.90(t,J＝6.8Hz,4H).
The fourth step compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (I) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-diisopropylethylamine (0.074 mL, 0.4476 mmol,2.0 eq) were added, followed by stirring at room temperature (18 ℃) for 0.5hr, and then decyl (ethyl) amine (63-3) (60 mg,0.324mmol, 1.45eq) was added and stirring was continued for 1hr. TLC (petroleum ether: ethyl acetate=5:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 16:1) to give the product N-decyl-3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-ethylpropionamide (63-4) (110 mg, 0.178 mmol, 80.12%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.36(m,2H),3.27(m,2H),3.21–3.14(m,1H),2.42(m,3H),2.34–2.17(m,2H),1.96(m,3H),1.76–1.65(m,3H),1.63(s,3H),1.61(m,1H),1.54(m,3H),1.27(d,J＝7.1Hz,15H),1.16(m,4H),1.08–0.97(m,2H),0.93(m,2H),0.88(m,10H),0.87(m,3H),0.86–0.83(m,2H),0.83–0.79(s,3H),0.76(s,3H),0.03(d,J＝2.7Hz,6H).
The fifth step compound N-decyl-3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-ethyl propionamide (63-4) (100 mg,0.162mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1.5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.627 mL,1.623mmol,10 eq) was added and then stirred at 50℃for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 45 ℃) to give a pale yellow oil, which was added with water (20 mL), extracted with ethyl acetate (20 ml×3), the organic phases were combined, washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give the product N-decyl-N-ethyl-3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionamide (63) (33.75 mg,0.058mmol, 35.66%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.46–3.16(m,5H),2.40(m,2H),2.27(m,1H),2.20(m,1H),1.95(m,5H),1.80–1.73(m,1H),1.67(m,2H),1.63(s,3H),1.61–1.48(m,3H),1.37(m,1H),1.26(s,15H),1.15(m,3H),1.10–1.01(m,2H),0.98(s,3H),0.87(m,5H),0.82(s,3H),0.80(s,3H).13C NMR(101MHz,CDCl3)δ172.33,132.44,127.95,123.84,79.09,77.32,77.21,77.00,76.68,54.35,54.01,50.86,47.64,45.53,42.34,40.58,38.83,38.36,36.98,36.53,33.49,32.17,31.88,29.60,29.54,29.29,28.05,27.66,22.66,21.95,21.79,19.51,15.36,14.28,14.09,13.04.LC-MS(ESI)[M+H]+＝502.7.
Example 64
Preparation of the Compound N- (2-aminopropyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionamide
Reference example 62 conversion of [ (2-aminoethyl) amino ] methanoic acid-2-methylpropan-2-yl ester to [ (2-aminopropyl) amino ] methanoic acid-2-methylpropan-2-yl ester gives the compound N- (2-aminopropyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (64).1H NMR(400MHz,DMSO)δ8.04-8.00(m,3H),3.09(tt,J＝13.9,7.0Hz,2H),3.03–2.97(m,1H),2.74(dd,J＝12.4,6.3Hz,2H),2.36–2.22(m,1H),2.22–1.80(m,6H),1.77–1.53(m,8H),1.53–1.18(m,5H),1.07–0.83(m,6H),0.82–0.72(m,5H),0.72–0.63(m,3H).13C NMR(101MHz,DMSO)δ172.81,132.63,127.29,77.38,54.40,53.99,49.04,38.93,37.94,37.03,36.55,35.93,35.16,33.51,32.18,28.66,27.90,27.79,25.59,22.06,21.88,19.76,16.28,14.58.LC-MS(ESI)[M+H]+＝391.2.
Example 65
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } -1- (nonylamino) methanamide
In the first step, the compound 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (200 mg,0.45mmol,1.0 eq) was dissolved in Toluene (toluene) (10 mL), diphenyl azide phosphate (128.88 mg,0.46mmol,1.05 eq) and triethylamine (54.16 mg) were added and the mixture was heated to 90℃to react for 1 hour. Cooled to 50 ℃, t-butanol (165.29 mg,2.23mmol,5.0 eq) and cuprous chloride (4.42 mg,0.04mmol,0.1 eq) were added and the temperature was raised to 90 ℃ for reaction for 12 hours. TLC (Petroleum ether: ethyl acetate=5:1) monitored completion of the reaction, concentrated reaction solution to remove toluene, diluted with water, extracted twice with ethyl acetate (20 mL), combined organic phases washed twice with brine, dried over anhydrous sodium sulfate, silica gel column chromatography (petroleum ether: ethyl acetate=50:10) to give ({ 2- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } amino) methanoic acid-2-methylpropan-2-yl ester (65-1) (231.70 mg, purity 90%, yield 46.61％).1H NMR(400MHz,CDCl3)δ4.47(s,1H),3.24–3.16(m,3H),3.01(s,1H),2.26(d,J＝7.4Hz,2H),1.99–1.91(m,2H),1.66–1.59(m,5H),1.45(s,6H),1.13–0.99(m,4H),0.89(s,9H),0.80(s,3H),0.75(s,2H),0.02(t,J＝7.5Hz,6H).
In the second step, the compound ({ 2- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) methanoic acid-2-methylpropan-2-yl ester (65-1) (120 mg,0.23mmol,1.0 eq) was dissolved in methanol hydrochloride (10 mL) and reacted at room temperature for 3 hours. TLC (dichloromethane: methanol=10:1,) monitored completion of the reaction, concentration of the reaction solution to remove methanol, silica gel column chromatography (dichloromethane: methanol=10:1) gave (4 aR,4 bS) -8- (2-aminoethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (65-2) (50.00 mg, purity 80%, yield) 56.72％).1H NMR(400MHz,DMSO)δ7.97(s,3H),3.46–3.40(m,1H),3.36(dd,J＝10.8,5.1Hz,2H),3.28(dd,J＝10.8,5.7Hz,2H),3.17(s,3H),2.51(s,3H),2.32(t,J＝8.2Hz,2H),1.90(t,J＝19.0Hz,3H),1.60(s,4H),0.88(s,3H),0.83–0.74(m,6H),0.69(s,3H).
In a third step, the compound non-1-amine (30 mg,0.209mmol,1.0 eq) was added to a reaction flask containing anhydrous dichloromethane (2 mL), N' -carbonyldiimidazole (67.91 mg, 0.319 mmol,2.0 eq) was added, and the mixture was stirred at room temperature (18 ℃) for 1hr. The reaction was monitored by LC-MS for completion. The reaction solution was dried under reduced pressure (water pump, 50 ℃ C.) to give the product N-nonylimidazole-1-carboxamide (65-3) (40 mg,0.169mmol, 80.48%) as a white solid. LC-MS (ESI) [ M+1] + =238.2 (Ms+1)
The fourth step compound N-nonylimidazole-1-carboxamide (65-3) (40 mg,0.169mmol,1.7 eq) was dissolved in anhydrous DMF (N, N-dimethylformamide) (2 mL), and (4 aR,4 bS) -8- (2-aminoethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (65-2) (30 mg,0.098mmol,1.0 eq) and N, N-Diisopropylethylamine (DIEA) (0.32 mL,0.196mmol,2.0 eq) were added and stirred at room temperature (18 ℃) for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction mixture was taken up in water (10 mL), extracted with ethyl acetate (15 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, the filtrate was dried under reduced pressure (water pump, 40 ℃ C.), and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product N- {2- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } -1- (nonylamino) methanamide (65) (21.85 mg,0.041mmol, 42.18%) as a white solid .1H NMR(400MHz,CDCl3)δ4.32(s,2H),3.33–2.97(m,5H),2.29(m,1H),2.19(d,J＝9.8Hz,1H),1.96(m,3H),1.81–1.66(m,4H),1.64(s,3H),1.57(m,1H),1.53–1.33(m,4H),1.27(s,12H),1.10–1.00(m,2H),0.97(s,3H),0.93–0.83(m,6H),0.80(d,J＝8.6Hz,6H).13C NMR(101MHz,CDCl3)δ158.30,130.01,129.67,79.06,77.32,77.21,77.01,76.69,54.27,53.90,40.78,39.27,38.82,37.94,36.96,36.51,33.59,32.22,31.85,30.22,29.53,29.35,29.25,28.05,27.64,26.92,22.65,21.89,21.84,19.75,15.36,14.27,14.08.LC-MS(ESI)[M+H]+＝475.4.
Example 66
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-hydroxypropyl) propionamide
Reference example 27 conversion of (1-methylindol-5-yl) methanamine to 3-hydroxypropionamine gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- (2-hydroxypropyl) propionamide (66).1H NMR(400MHz,CDCl3)δ5.90(s,1H),3.62(t,J＝5.4Hz,2H),3.49–3.34(m,2H),3.22(dd,J＝11.4,4.6Hz,1H),2.51–2.41(m,1H),2.32-2.26(m,2H),2.19-2.22(m,3H),2.08–2.04(m,1H),2.03–1.97(m,1H),1.95–1.87(m,2H),1.80–1.73(m,1H),1.70–1.65(m,4H),1.61-1.55(m,4H),1.42–1.32(m,1H),1.08–1.00(m,2H),0.98-0.97(m,3H),0.90–0.84(m,2H),0.81-0.79(m,6H).13C NMR(101MHz,CDCl3)δ174.34,131.61,128.68,79.04,59.12,54.30,54.02,53.93,38.83,37.89,36.94,36.51,36.17,35.54,33.55,32.43,32.11,28.04,27.62,25.22,21.91,21.78,19.54,15.36,14.27.LC-MS(ESI)[M+H]+＝392.2.
Example 67
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (4-methylpiperazin-1-yl) hexyl ] propanamide
The first-step compound [ (6-bromohexyl) amino ] methanoic acid-2-methylpropan-2-yl ester (300 mg,1.07mmol,1.0 eq)) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (3 mL), cesium carbonate (872 mg,2.67mmol,2.5 eq)) and 1-methylpiperazine (128 mg,1.28mmol,1.2 eq)) were added, and then stirred at room temperature (20 ℃) for 16hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. Water (15 mL) was added, extraction was performed with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil, the crude product was purified by column chromatography (dichloromethane: methanol=1:0 to 6:1), separation and collection of new spots (R f =0.35) to give the product { 6- (4-methylpiperazin-1-yl) hexyl ] amino } propan-2-yl-methanoate (67-1) (240 mg,0.80 mmol, 74.8%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + =300.2
The second step compound { [6- (4-methylpiperazin-1-yl) hexyl ] amino } methanoic acid-2-methylpropan-2-yl ester (67-1) (90 mg,0.301mmol,1.0 eq) was added to a reaction flask containing methanol (0.5 mL), and after adding a hydrogen chloride methanol solution (0.093 mL,3.00mmol,10 eq)), stirred at room temperature (18 ℃) for 2hr. LC-MS monitored the reaction. After the reaction solution was dried under reduced pressure (water pump, 50 ℃ C.) and replaced with oil pump for 0.2hr, 6- (4-methylpiperazin-1-yl) hex-1-amine (67-2) (60 mg,0.271mmol, 90.1%) was obtained as a white solid. LC-MS (ESI) [ m+h ] + =200.2
The third step compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (67-2) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (110 mg,0.290mmol,1.3 eq) and DIEA (N, N-diisopropylethylamine) (0.074 mL,0.446mmol,2.0 eq) was added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by addition of 6- (4-methylpiperazin-1-yl) hex-1-amine (57.7 mg,0.290mmol,1.3 eq) and stirring continued for 1hr. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃) and water (15 mL) was added, extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil. The crude product was purified by column chromatography (dichloromethane: methanol=1:0 to 16:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (4-methylpiperazin-1-yl) hexyl ] propionamide (67-3) (80 mg,0.127mmol, 56.9%) as a pale yellow solid. LC-MS (ESI) [ m+h ] + =630.5
The fourth step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (4-methylpiperazin-1-yl) hexyl ] propanamide (67-3) (80 mg,0.127mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.27 mL,1.27mmol,10 eq) was added and then heated to 50℃and stirred for 16hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil. Water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 23:2) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (4-methylpiperazin-1-yl) hexyl ] propanamide (67) (10.9 mg,0.020mmol, 15.6%) as a white solid .1H NMR(400MHz,CDCl3)δ5.49(s,1H),3.22(d,J＝6.6Hz,3H),2.64(s,6H),2.43(m,3H),2.37(s,3H),2.34–2.13(m,3H),1.97(m,5H),1.82–1.68(m,3H),1.64(s,3H),1.59–1.45(m,5H),1.33(s,5H),1.25(s,2H),1.05(m,2H),0.98(s,3H),0.86(m,3H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ172.89,131.90,128.45,79.04,77.33,77.22,77.02,76.70,58.07,54.29,54.09,53.93,52.23,45.35,39.36,38.82,37.90,36.93,36.50,35.60,33.55,32.11,29.56,28.05,27.63,27.01,26.66,26.09,25.16,21.90,21.79,21.42,19.58,15.37,14.29.LC-MS(ESI)[M+H]+＝516.4.
Example 68
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-3-yl) heptyl ] propionamide
In the first step, n-butyllithium (25.480 mL,64.426mmol,1.2 eq) was added dropwise to a reaction flask containing diisopropylamine (6.52 g,64.426mmol,1.2 eq) and anhydrous tetrahydrofuran (20 mL) at-68℃and stirred for 1hr, then a solution of 3-methylpyridine (5.00 g,53.688mmol,1.0 eq) in tetrahydrofuran (20 mL) was added and stirring was continued at-68℃for 1hr, and then 6-bromohex-1-ol (12.64 g,69.795mmol,1.3 eq) was added and stirred at room temperature (15 ℃) for 16hr; TLC (dichloromethane: methanol=20:1) monitored that most of the reaction was starting material, new spots with polarity increased were formed, after quenching the reaction with aqueous saturated ammonium chloride solution (10 mL), water (10 mL) was added, extracted with ethyl acetate (20 ml×3), the organic phases were combined, washed with water (20 ml×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), yielding a yellow liquid; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 24:1) to give the product 7- (pyridin-3-yl) heptan-1-ol (68-1) (750 mg,1.940mmol, 3.61%) as a pale yellow oil. LC-MS (ESI) [ m+1] + =194.1
In a second step, 7- (pyridin-3-yl) heptan-1-ol (68-1) (750 mg, 3.750 mmol,1.0 eq) was added to a reaction flask containing acetonitrile (20 mL) and triphenylphosphine (2035.46 mg,7.760mmol,2.0 eq) and carbon tetrabromide (2573.57 mg,7.760mmol,2.0 eq) were added at 0deg.C and stirring was continued after returning to room temperature (18 ℃ C.) for 1hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was taken up in water (20 mL), extracted with dichloromethane (20 mL x 3), the organic phases combined, dried under reduced pressure (water pump, 45 ℃) to give crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give 3- (7-bromoheptyl) pyridine (68-2) (220 mg,0.809mmol, 20.84%) as a colourless oil .1H NMR(400MHz,CDCl3)δ8.45–8.44(m,2H),7.54-7.53(d,J＝7.7Hz,1H),7.26–7.20(m,1H),3.40(t,J＝6.8Hz,2H),2.65–2.59(m,2H),1.89–1.80(m,2H),1.62(dd,J＝14.7,7.1Hz,2H),1.48–1.40(m,2H),1.39–1.32(m,4H).
The third step compound 3- (7-bromoheptyl) pyridine (68-2) (220 mg,0.859mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (3 mL), potassium phthalimide (175.90 mg,0.945mmol,1.1 eq) was added and stirred at room temperature (15 ℃) for 16hr, and TLC (petroleum ether: ethyl acetate=1:1) monitored for completion. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with water (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give the product 2- [7- (pyridin-3-yl) heptyl ] isoindole-1, 3-dione (68-3) (210 mg,0.616mmol, 71.75%) as a colorless oil .1H NMR(400MHz,CDCl3)δ8.46(dd,J＝20.1,5.3Hz,2H),7.84(dt,J＝7.1,3.5Hz,2H),7.75–7.68(m,2H),7.51(d,J＝7.7Hz,1H),7.22(dd,J＝7.7,4.9Hz,1H),3.68(t,J＝7.3Hz,2H),2.67–2.55(m,2H),1.72–1.56(m,4H),1.36(s,6H).
The fourth step compound 2- [7- (pyridin-3-yl) heptyl ] isoindole-1, 3-dione (68-3) (120 mg,0.372mmol,1.0 eq) was added to a reaction flask containing ethanol (3 mL), hydrazine hydrate (0.045 mL,0.744mmol,2.0 eq) was added and the mixture was heated to 80℃and stirred for 16hr; the reaction was monitored by LC-MS for completion. The reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) and then was substituted by oil pump to give 7- (pyridin-3-yl) hept-1-amine (68-4) (70 mg,0.328mmol, 88.01%) as a pale yellow oil. LC-MS (ESI) [ m+1] + =193.1
The fifth step compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (I) (100 mg,0.223mmol,1.0 eq)) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (101.67 mg,0.267mmol,1.2 eq) and N, N-diisopropylethylamine (0.074 mL, 0.4476 mmol,2.0 eq) were added and stirred at room temperature (18 ℃) for 0.5hr, followed by 7- (pyridin-3-yl) hept-1-amine (68-4) (55.71 mg,0.290mmol,1.3 eq) and stirring continued for 1hr. TLC (dichloromethane: methanol=20:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- [7- (pyridin-3-yl) heptyl ] propionamide (68-5) (90 mg,0.109mmol, 48.79%) as a pale yellow oil. LC-MS (ESI) [ m+1] + = 623.3
The sixth step 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-3-yl) heptyl ] propanamide (68-5) (90 mg,0.144mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (1.445 mL,1.445mmol,10 eq) was added and the mixture was warmed to 50℃and stirred for 16hr. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-3-yl) heptyl ] propionamide (68) (37.10 mg,0.070mmol, 48.50%) as a white solid .1H NMR(400MHz,CDCl3)δ8.46(d,J＝5.3Hz,2H),7.55(d,J＝7.8Hz,1H),7.27(t,J＝3.7Hz,1H),5.51(s,1H),3.22(dd,J＝11.7,4.7Hz,3H),2.62(t,J＝7.7Hz,2H),2.48–2.26(m,2H),2.25–2.17(m,1H),2.15–2.00(m,2H),1.99–1.84(m,3H),1.80–1.74(m,1H),1.72–1.63(m,4H),1.62(s,3H),1.57(m,2H),1.48(m,2H),1.31(s,J＝16.9Hz,7H),1.10–1.00(m,2H),0.97(s,3H),0.88(ddd,J＝11.8,9.1,5.9Hz,3H),0.80(d,J＝7.3Hz,6H).13C NMR(101MHz,CDCl3)δ172.86,149.50,146.84,138.36,136.67,131.93,128.43,123.62,79.01,77.34,77.22,77.02,76.70,54.31,53.95,39.45,38.83,37.91,36.95,36.51,35.61,33.57,32.93,32.12,30.92,29.66,29.02,28.95,28.05,27.64,26.79,25.15,21.91,21.79,19.56,15.37,14.28.LC-MS(ESI)[M+H]+＝509.4.
Example 69
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [6- (piperazin-1-yl) hexyl ] propanamide
In the first step, 6-bromohexan-1-ol (2.0 g,11.0mmol,1.0 eq) and potassium phthalimide (2.5 g,13.3mmol,1.2 eq) were dissolved in DMF (N, N-dimethylformamide) (15.0 mL) and the reaction stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (50 mL. Times.2), and washed with water (60 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was subjected to column chromatography using petroleum ether ethyl acetate=5:1 to give 2- (6-hydroxyhexyl) isoindole-1, 3-dione (69-1) (2.30 g, purity 90%, yield 75.8%). LC-MS (ESI) [ m+h ] +＝248.1,tR =1.068 min.
The second step compound 2- (6-hydroxyhexyl) isoindole-1, 3-dione (69-1) (750 mg, 3.750 mmol,1.0 eq) was added to a reaction flask containing acetonitrile (20 mL) and triphenylphosphine (2035.46 mg,7.760mmol,2.0 eq) and carbon tetrabromide (2573.57 mg,7.760mmol,2.0 eq) were added at 0deg.C and stirred for a further 1hr at room temperature (18deg.C). TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction mixture was taken up in water (20 mL), extracted with dichloromethane (20 mL x 3), the organic phases combined, dried under reduced pressure (water pump, 45 ℃) to give the crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give the product 2- (6-hydroxyhexyl) isoindole-1, 3-dione (69-2) (220 mg,0.809mmol, 20.84%) as a colourless oil.
In the third step, 2- (6-hydroxyhexyl) isoindole-1, 3-dione (69-2) (120 mg,0.39mmol,1.0 eq), potassium carbonate (160.4 mg,1.16mmol,3.0 eq) and piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (114.3 mg,0.46mmol,1.2 eq) were dispersed in N, N-dimethylformamide (5 mL), and the reaction mixture was stirred at room temperature for 18hrs. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (50 mL) and washed with water (40 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was subjected to column chromatography with petroleum ether: ethyl acetate=20:1-10:1 to give 4- [6- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) hexyl ] piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (69-3) (110 mg, purity 90%, yield 61.1%).
LC-MS(ESI)[M+H]+＝416.2.
In the fourth step, 4- [6- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) hexyl ] piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (69-3) (0.5 g,1.20mmol,1.0 eq) was dissolved in ethanol (20 mL), hydrazine hydrate (0.29 mL,6.02mmol,5.0 eq) was added, the reaction was stirred at 80℃for 2H, and TLC (dichloromethane: methanol=10:1) monitored for completion. After completion of the reaction, the mixture was concentrated, extracted with chloroform (20 mL), washed with water (20 mL. Times.2), and dried over anhydrous Na 2SO4 to obtain a crude product (69-4) which was directly taken into the next step.
In a fifth step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min, the reaction 4- (6-aminohexyl) piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (69-4) (82.7 mg,0.29mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude was purified using dichloromethane methanol=15:1 to give 4- [6- ({ 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) hexyl ] piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (69-5) (110 mg, purity 90%, yield 62.0%) and the product was taken directly to the next step.
In the sixth step, 4- [6- ({ 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) hexyl ] piperazine-1-carboxylic acid 2-methylpropan-2-yl ester (69-5) (90 mg,0.13mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), a solution of TBAF (1.26 mL,1.26mmol,10.0 eq) in tetrahydrofuran was added, and the reaction mixture was stirred overnight at 60 ℃. TLC (dichloromethane: methanol=10:1) monitored the reaction. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate and the crude was concentrated in dichloromethane: methanol=10:1, followed by column chromatography to give 4- [6- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) hexyl ] piperazine-1-carboxylic acid 2-methylpropan-2-yl ester (69-6) (50 mg, purity 90%, yield 59.5%).
In the seventh step, 4- [6- ({ 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) hexyl ] piperazine-1-carboxylic acid 2-methylpropan-2-yl ester (69-6) (50 mg,0.07mmol,1.0 eq) was dissolved in a methanol solution of hydrochloric acid (1 mol/L,2 mL), and the reaction mixture was stirred at room temperature for 2hrs. The reaction was monitored by TLC (petroleum ether: ethyl acetate=5:1). The crude product was concentrated by passing through a column with methylene chloride, methanol=50:1-10:1, to give 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (piperazin-1-yl) hexyl ] propionamide (69) (16.2 mg, purity 91.7%, yield) as a white solid after concentration 35.3％).1H NMR(400MHz,D2O)δ3.38(s,4H),3.17-3.00(m,7H),2.90–2.78(m,2H),2.42(s,1H),2.29–2.00(m,4H),1.89-1.80(m,3H),1.65-1.55(m,10H),1.43(s,3H),1.29(s,5H),1.10-0.88(m,6H),0.76-0.72(m,8H).13C NMR(101MHz,DMSO)δ172.17,132.80,127.11,77.38,54.41,54.00,48.85,40.65,40.44,40.23,40.02,39.82,39.61,39.40,38.92,38.76,37.91,37.07,36.55,35.20,33.56,32.17,29.51,28.67,27.90,26.58,26.55,25.54,22.06,21.87,19.77,16.28,14.58.LC-MS(ESI)[M+H]+＝502.4.
Example 70
Preparation of the Compound N- [9- (dimethylamino) nonyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
In the first step, 9-bromononan-1-ol (2.0 g,11.0mmol,1.0 eq) and potassium phthalimide (2.5 g,13.3mmol,1.2 eq) were dissolved in DMF (N, N-dimethylformamide) (15.0 mL), and the reaction mixture was stirred at room temperature overnight. TLC (petroleum ether: ethyl acetate=5:1) monitored the reaction. The reaction mixture was extracted with ethyl acetate (50 mL. Times.2), and washed with water (60 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and concentrated and the crude product was purified by passing it over petroleum ether ethyl acetate=5:1 to give 2- (9-hydroxynonyl) isoindole-1, 3-dione (70-1) which was used directly in the next step.
In a second step, 2- (9-hydroxynonyl) isoindole-1, 3-dione (70-1) (3.0 g,10.37mmol,1.0 eq) was dissolved in dichloromethane (60 mL) followed by addition of carbon tetrabromide (6.88 g,20.73mmol,2.0 eq), triphenylphosphine (5.44 g,20.73mmol,2.0 eq) and the reaction stirred at room temperature for 16hrs, and TLC (petroleum ether: ethyl acetate=10:1) monitored for reaction completion. After concentration, the crude product was purified with petroleum ether: ethyl acetate=20:1 was passed through a column to give 2- (9-bromononyl) isoindole-1, 3-dione (70-2) (2.6 g, purity 90%, yield 64.1%). LC-MS (ESI) [ m+na ] + =376.0
In the third step, 2- (9-bromononyl) isoindole-1, 3-dione (70-2) (1.0 g,2.84mmol,1.0 eq), potassium carbonate (1.18 g,8.52mmol,3.0 eq), dimethylamine hydrochloride (0.35 g,4.26mmol,1.5 eq) were dispersed in N, N-dimethylformamide (10 mL), the reaction was stirred at room temperature for 18hrs, and TLC (dichloromethane: methanol=10:1) monitored for completion. The reaction mixture was extracted with ethyl acetate (50 mL) and washed with water (40 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude product after concentration was purified by passing through a column with methylene chloride: methanol=20:1-10:1 to give 2- [9- (dimethylamino) nonyl ] isoindole-1, 3-dione (70-3) (0.5 g, purity 90%, yield 50.0％).1H NMR(400MHz,CDCl3)δ7.89–7.79(m,2H),7.75–7.66(m,2H),3.72–3.62(m,2H),2.63–2.51(m,2H),2.46(s,6H),1.66(dd,J＝14.5,7.1Hz,4H),1.31(d,J＝9.0Hz,10H).
In the fourth step, 2- [9- (dimethylamino) nonyl ] isoindole-1, 3-dione (70-3) (0.3 g,0.95mmol,1.0 eq) was dissolved in ethanol (20 mL), hydrazine hydrate (0.23 mL,5.74mmol,5.0 eq) was added, the reaction was stirred at 80℃for 2h, and TLC (dichloromethane: methanol=10:1) monitored to be complete. Concentrating after the reaction, extracting with chloroform (20 mL), washing with water (20 mL. Times.2), drying with anhydrous Na 2SO4 to obtain the crude product of 9- (dimethylamino) non-1-amine (70-4), and directly feeding into the next step .1H NMR(400MHz,DMSO)δ2.53(t,J＝6.7Hz,2H),2.18–2.11(m,2H),2.08(s,6H),1.40-135(m,4H),1.24(s,10H).
In a fifth step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min, the reaction 9- (dimethylamino) non-1-amine (70-4) (62.3 mg,0.35mmol,1.5 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was purified by passing methanol=15:1 through a column to give N- [9- (dimethylamino) nonyl ] -3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (70-5) (80 mg, purity 90%, yield 52.3%) and the product was directly taken to the next step.
In the sixth step, N- [9- (dimethylamino) nonyl ] -3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (70-5) (80 mg,0.13mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), and a solution of TBAF (1.29 mL,1.29mmol,10.0 eq) in tetrahydrofuran was added and the reaction mixture was stirred overnight at 60 ℃. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate and the crude product was purified by column chromatography with dichloromethane, methanol=10:1, and concentrated to give N- [9- (dimethylamino) nonyl ] -3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (70) as a white solid (10 mg, purity 83.9%, yield 12.8％).1H NMR(400MHz,CDCl3)δ5.502-5.42(m,1H),3.24-3.20(m,3H),2.62–2.53(m,2H),2.46(s,6H),2.42–2.06(m,5H),2.07-1.87(m,3H),1.80–1.54(m,10H),1.51–1.42(m,2H),1.37(dd,J＝13.3,3.5Hz,1H),1.27-1.25(m,10H),1.19–0.95(m,6H),0.95–0.74(m,9H).13C NMR(101MHz,CDCl3)δ172.84,131.95,128.45,79.04,77.32,77.00,76.69,58.90,54.32,53.97,44.07,39.48,38.83,37.92,36.96,36.52,35.63,33.58,32.13,29.64,29.24,29.18,29.10,28.05,27.65,27.03,26.82,25.97,25.15,21.92,21.80,19.56,15.36,14.29.LC-MS(ESI)[M+H]+＝503.4.
Example 71
Preparation of the Compound N- (hexahydropyridin-4-yl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
Reference example 62 conversion of [ (2-aminoethyl) amino ] methanoic acid-2-methylpropan-2-yl ester to 4-aminocyclopyrachlor-1-carboxylic acid-2-methylpropan-2-yl ester gives the compound N- (hexahydropyridin-4-yl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (71).1H NMR(400MHz,CDCl3)δ5.82(d,J＝7.7Hz,1H),4.05(m,1H),3.51(d,J＝12.4Hz,2H),3.22(dd,J＝11.4,4.4Hz,1H),2.98(t,J＝11.8Hz,2H),2.29(m,3H),2.19–2.07(m,4H),2.01(m,2H),1.90(m,3H),1.76(m,2H),1.71–1.65(m,3H),1.63(s,3H),1.59(m,1H),1.43–1.30(m,2H),1.04(dd,J＝13.6,9.4Hz,2H),0.99–0.93(s,3H),0.85(m,2H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ172.64,131.59,128.79,79.04,77.33,77.22,77.02,76.70,54.26,53.90,44.06,43.28,38.83,37.88,37.46,36.90,36.51,35.37,33.54,32.12,31.69,28.77,28.06,27.62,21.89,21.81,19.64,15.39,14.31.LC-MS(ESI)[M+H]+＝417.
Example 72
Preparation of the Compound N- (9-cyanononyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
The first step compound 2- (9-bromononyl) isoindole-1, 3-dione (70-2) (300 mg, 0.850 mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), potassium cyanide (110 mg,1.68mmol,1.98 eq) was added, then heated to 120 ℃ and stirred for 16hr, and tlc (petroleum ether: ethyl acetate=5:1) monitored to complete the reaction. The reaction mixture was taken up in water (15 mL), extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with water (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give the product 10- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) decanol (72-1) (55 mg,0.184mmol, 21.6%) as a yellow oil .1H NMR(400MHz,CDCl3)δ7.84(dt,J＝7.1,3.5Hz,2H),7.75–7.68(m,2H),3.74–3.62(m,2H),2.33(t,J＝7.2Hz,2H),1.64(d,J＝7.4Hz,4H),1.43(dd,J＝10.0,4.6Hz,2H),1.32(dd,J＝7.2,4.0Hz,8H).
The second step of the compound 10- (1, 3-dioxy-2, 3-dihydro-1H-isoindol-2-yl) decanitrile (72-1) (55 mg,0.184mmol,1.0 eq) was added to a reaction flask containing ethanol (2 mL), hydrazine hydrate (0.018 mL,0.369mmol,2.0 eq) was added, the mixture was stirred at 80℃for 16hr, the reaction was monitored by LC-MS and completed, the reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a white viscous substance, which was dried by oil pump instead of oil to give crude 10-aminodecanol (72-2) (30 mg,0.178mmol, 96.7%) as a white viscous substance. LC-MS (ESI) [ m+h ] + =169.1
In a third step, compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (I) (30 mg,0.067mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (1 mL), HATU (33.04 mg,0.087mmol,1.3 eq) and DIEA (N, N-diisopropylethylamine) (0.022 mL,0.134mmol,2.0 eq) were added and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by 10-aminodecylnitrile (72-2) (22.50 mg,0.134mmol,2.0 eq) and stirring continued for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. The reaction solution was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil, and the crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 4:1) to give the product N- (9-cyanononyl) -3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionamide (72-3) (30 mg,0.050mmol, 74.9%) as a colorless oil .1H NMR(400MHz,CDCl3)δ5.47(d,J＝4.3Hz,1H),3.23(d,J＝4.5Hz,2H),3.20–3.14(m,1H),2.42(d,J＝8.7Hz,1H),2.34(t,J＝7.1Hz,3H),2.26(m,1H),2.14(m,3H),2.05–1.75(m,8H),1.58–1.39(m,8H),1.30(s,10H),1.05–0.98(m,2H),0.88(d,J＝5.4Hz,12H),0.84(d,J＝8.0Hz,2H),0.80(s,3H),0.75(s,3H),0.03(d,J＝2.8Hz,6H).
The fourth step compound N- (9-cyanononyl) -3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (72-3) (30 mg,0.050mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (0.501 mL,0.501mmol,10 eq), the mixture was warmed to 50℃and stirred for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil, which was extracted with ethyl acetate (10 ml×3) and the organic phases were combined, washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give the product N- (9-cyanononyl) -3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (72) (9 mg,0.018mmol, 35.2%) as a white solid .1H NMR(400MHz,CDCl3)δ5.45(s,1H),3.23-3.22(d,J＝6.6Hz,3H),2.42-2.40(m,1H),2.32-2.24(dd,J＝19.2,12.2Hz,3H),2.17-2.08(m,3H),1.96-1.88(m,3H),1.78-1.75(d,J＝13.2Hz,2H),1.66-1.64(m,4H),1.63(s,3H),1.59(m,2H),1.46-1.38(m,4H),1.30(s,9H),1.08–1.00(m,2H),0.95(s,3H),0.86-0.85(m,2H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ172.87,131.91,128.48,119.83,79.05,77.34,77.03,76.71,54.28,53.93,39.48,38.83,37.89,36.93,36.51,35.61,33.57,32.11,29.68,29.17,29.11,28.67,28.59,28.05,27.62,26.84,25.32,25.15,21.90,21.79,19.58,17.13,15.38,14.29.LC-MS(ESI)[M+H]+＝485.35.
Example 73
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -1- (piperazin-1-yl) propan-1-one
In the first step, piperazine (2.0 g,23.2mmol,1.0 eq) was dissolved in methylene chloride (30 mL), and triethylamine (4.0 mL,29.0mmol,5.0 eq), (BOC) 2 O (1.3 mL,5.8mmol,0.4 eq) was added and the reaction mixture was stirred at room temperature for 16h. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was washed with saturated ammonium chloride (30 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with methylene chloride: methanol=20:1, and concentrated to give piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-1) (0.5 g, purity 90%, yield 41.6%) as a colorless liquid. 1H NMR(400MHz,CDCl3 ) Delta 3.47-3.36 (m, 4H), 2.88-2.79 (m, 4H), 1.46 (s, 9H).
In the second step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (120 mg,0.27mmol,1.0 eq), HATU (122.0 mg,0.32mmol,1.2 eq) and N, N-diisopropylethylamine (44.9 mg,0.35mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min, the reaction piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-1) (98.8 mg,0.40mmol,1.5 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate=3:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 3:1 passing through a column gives 4- {3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-2) (110 mg, purity 90%, yield 60%) the product was directly fed to the next step.
In the third step, 4- {3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-2) (110 mg,0.19mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), and a tetrahydrofuran solution of TBAF (1.90 mL,1.94mmol,10.0 eq) was added thereto, and the reaction mixture was stirred overnight at 60 ℃. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude was purified by column chromatography with dichloromethane: methanol=20:1 to give 4- {3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-3) (90 mg, purity 90%, yield 82.8%) as a white solid.
In the fourth step, 4- {3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } piperazine-1-carboxylic acid-2-methylpropan-2-yl ester (73-3) (60 mg,0.12mmol,1.0 eq) was dissolved in a hydrogen chloride-methanol solution (4 mol/L,2 mL), and the reaction mixture was stirred at room temperature for 2hrs. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The crude product was concentrated by passing through a column with methylene chloride, methanol=20:1-10:1, to give 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1- (piperazin-1-yl) propan-1-one (73) (21.5 mg, purity 91.3%, yield) as a white solid after concentration 40.9％).1H NMR(400MHz,CDCl3)δ3.93(s,2H),3.80(s,2H),3.24-3.19(m,5H),2.37-2.35(m,2H),2.25-2.20(m,1H),2.11(d,J＝13.9Hz,1H),2.05–1.85(m,3H),1.81–1.64(m,4H),1.64–1.53(m,4H),1.42-1.37(m,1H),1.25(s,2H),1.17-1.08(m,3H),0.98-0.94(m,3H),0.91–0.76(m,8H).13C NMR(101MHz,CDCl3)δ171.46,131.69,128.53,79.03,77.23,54.28,53.90,38.83,38.19,36.92,36.51,33.44,32.17,32.09,29.70,28.08,27.61,27.41,24.57,21.89,21.73,21.40,19.57,15.41,14.30.LC-MS(ESI)[M+H]+＝417.25.
Example 74
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (7-phenylheptyl) propionamide
Reference example 43 conversion of m-fluorobenzaldehyde to benzaldehyde gave the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- (7-phenylheptyl) propanamide (74).1H NMR(400MHz,CDCl3)δ7.30–7.26(m,2H),7.19-7.16(m,3H),5.47(s,1H),3.22(dd,J＝11.5,4.7Hz,3H),2.68–2.53(m,2H),2.48–2.39(m,1H),2.38–2.20(m,2H),2.19–2.08(m,2H),2.05–1.86(m,3H),1.79–1.73(m,2H),1.73–1.65(m,3H),1.62(s,3H),1.61-1.56(m,1H),1.52–1.37(m,3H),1.33-1.32(m,5H),1.30-1.25(m,1H),1.15–0.98(m,3H),0.97(s,3H),0.92(dd,J＝12.5,4.1Hz,1H),0.89–0.83(m,2H),0.81-0.79(m,6H).13C NMR(101MHz,CDCl3)δ172.89,142.75,131.90,128.51,128.39,128.25,125.61,79.05,77.34,77.02,76.70,54.29,53.94,39.53,38.83,37.88,36.94,36.51,35.93,35.57,33.57,32.11,31.40,29.67,29.18,28.05,27.64,26.87,25.14,21.91,21.80,19.57,15.37,14.29.LC-MS(ESI)[M+H]+＝508.
Example 75
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (3, 4,5, 6-tetrahydro-2H-pyran-4-ylmethyl) propanamide
Reference example 27 conversion of (1-methylindol-5-yl) methanamine to 3,4,5, 6-tetrahydro-2H-pyran-4-ylmethylamine gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3, 4,5, 6-tetrahydro-2H-pyran-4-ylmethyl) propanamide (75).1H NMR(400MHz,CDCl3)δ5.54(s,1H),3.97(dd,J＝11.3,3.4Hz,2H),3.36(t,J＝11.0Hz,2H),3.22(dd,J＝11.5,4.6Hz,1H),3.19–3.09(m,2H),2.56–2.39(m,1H),2.39–2.21(m,2H),2.21–2.09(m,2H),2.07–1.85(m,3H),1.76(m,2H),1.73–1.69(m,2H),1.67(m,2H),1.63(s,3H),1.62–1.55(m,3H),1.37(m,1H),1.30(dd,J＝12.5,3.9Hz,2H),1.08–1.00(m,2H),0.98(s,3H),0.89–0.84(m,2H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ173.08,131.77,128.58,79.04,77.34,77.22,77.02,76.70,67.60,54.28,53.91,45.14,38.83,37.85,36.92,36.51,35.56,35.31,33.57,32.12,30.60,28.05,27.62,25.13,21.90,21.76,19.59,15.37,14.28.LC-MS(ESI)[M+H]+＝432.
Example 76
Preparation of the Compound N-decyl-3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-hydroxyethyl) propanamide
The first step compound decan-1-amine (300 mg, 1.227 mmol,1.0 eq) was added to a reaction flask containing acetonitrile (2 mL), 1-bromo-4, 5-tetramethyl-3-oxa-4-sila-ne (547.51 mg,2.289mmol,1.2 eq) and K2CO3 (342.67 mg,2.479mmol,1.3 eq) were added, and after stirring was performed at 80 ℃ for 4hr, tlc (dichloromethane: methanol=10:1) was monitored to be substantially complete. The reaction solution is directly stirred with silica gel, and is separated and purified by column chromatography (dichloromethane: methanol=1:0 to 19:1) to obtain the product 2, 3-tetramethyl-7-aza-4-oxa-3-silaheptadecane (76-1) (30 mg,0.095mmol, 29.9%) as light yellow oily substance .1H NMR(400MHz,CDCl3)δ3.74(t,J＝5.3Hz,2H),2.77(t,J＝5.3Hz,2H),2.68(t,J＝7.4Hz,2H),1.52(d,J＝7.2Hz,2H),1.20(d,J＝17.2Hz,14H),0.84–0.76(m,12H),-0.00(s,6H).
The second step compound 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (2 mL), HATU (74.56 mg,0.196mmol,1.1 eq) and DIEA (N, N-diisopropylethylamine) (0.059 mL, 0.356 mmol,2.0 eq) were added and stirred for 0.5hr, and 2, 3-tetramethyl-7-aza-4-oxa-3-silaheptadecane (76-1) (61.89 mg,0.196mmol,1.1 eq) was added and stirring was continued at room temperature (18 ℃) for 1hr. TLC (petroleum ether: ethyl acetate=5:1) monitored reaction completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 16:1), the product N-decyl-3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a is obtained by separation and purification-dodecahydrophenanthrene ] -N- (4, 5-tetramethyl-3-oxa-4-silahex-1-yl) propanamide (76-2) (100 mg,0.134mmol, 75.1%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.70(m,2H),3.27(m,4H),3.13(dd,J＝11.2,4.6Hz,1H),2.32(m,5H),1.97–1.82(m,3H),1.68–1.42(m,12H),1.21(t,J＝7.0Hz,18H),0.93(m,4H),0.84(s,19H),0.73(dd,J＝20.9,4.4Hz,7H),-0.01(dd,J＝6.0,2.3Hz,12H).
The third step compound N-decyl-3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydro-gen phenanthryl ] -N- (4, 5-tetramethyl-3-oxa-4-silahex-1-yl) propanamide (76-2) (100 mg,0.134mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (1.5 mL), after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (1.34 mL,1.34mmol,10 eq) the mixture was heated to 50℃and stirred for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. The reaction mixture was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil. Water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:3) to give the product N-decyl-3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (2-hydroxyethyl) propanamide (76) (21.3 mg,0.036mmol, 26.9%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.76(d,J＝4.1Hz,2H),3.64–3.47(m,2H),3.47–3.26(m,2H),3.22(dd,J＝11.5,4.5Hz,1H),2.50–2.12(m,6H),1.93(t,J＝15.9Hz,3H),1.80–1.49(m,10H),1.45–1.19(m,16H),1.13–1.02(m,2H),0.98(S,3H),0.88(t,J＝6.6Hz,4H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ175.47,175.47,132.03,128.36,79.06,77.34,77.02,76.70,62.96,54.32,53.95,50.22,49.79,38.83,38.32,36.95,36.51,33.46,32.27,32.19,31.87,29.57,29.53,29.38,29.29,29.16,28.06,27.63,26.85,25.11,22.67,21.93,21.75,19.53,15.38,14.27,14.11.LC-MS(ESI)[M+H]+＝518.40.
Example 77
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-methoxyethyl) propanamide
Reference example 27 conversion of (1-methylindol-5-yl) methanamine to 2-methoxyethylamine gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- (2-methoxyethyl) propanamide (77).1H NMR(400MHz,CDCl3)δ5.88(s,1H),3.45(s,4H),3.36(s,3H),3.26–3.19(m,1H),2.53–2.21(m,2H),2.17-2.14(m,2H),2.04–1.86(m,3H),1.77(dd,J＝9.7,6.3Hz,1H),1.72–1.52(m,7H),1.42-1.20(m,2H),1.06 -1.02(m,2H),1.00-0.97(m,3H),0.91(d,J＝4.1Hz,1H),0.89–0.83(m,2H),0.82-0.80(m,6H).13C NMR(101MHz,CDCl3)δ172.99,131.76,128.60,79.06,77.34,77.03,76.71,71.29,58.75,54.29,53.95,39.14,38.83,37.86,36.94,36.51,35.44,34.94,33.58,32.10,28.05,27.63,25.03,21.90,21.75,19.54,15.37,14.28.LC-MS(ESI)[M+H]+＝392.
Example 78
Preparation of decyl 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionate
In the first step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq) was dissolved in methylene chloride (5 mL), oxalyl chloride (0.1 mL,0.12mmol,1.4 eq) was added and stirred at room temperature for 0.5h, then methylene chloride and excess oxalyl chloride were removed by suspension evaporation, and then dissolved in methylene chloride (5 mL), triethylamine (0.06 mg,0.45mmol,5.0 eq) and decanol (15.5 mg,0.10mmol,1.1 eq) were added, and the reaction mixture was stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=10:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=15:1 was passed through a column to give decyl 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionate (78-1) (25 mg, purity 90%, yield 42.8%), the product was directly taken to the next step.
In the second step, decyl 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionate (78-1) (45 mg,0.08mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), a solution of TBAF (0.76 mL,0.76mmol,10.0 eq) in tetrahydrofuran was added, and the reaction mixture was stirred overnight at 50 ℃. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with petroleum ether ethyl acetate=10:1 to give decyl 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionate (78-2) (4.9 mg, purity 85.4%, yield) as a white solid after concentration 11.7％).1H NMR(400MHz,CDCl3)δ4.09–4.02(m,2H),3.22(dd,J＝11.4,4.7Hz,1H),2.49–2.38(m,1H),2.37–2.32(m,1H),2.30–2.20(m,1H),2.18–2.10(m,1H),2.06–1.84(m,3H),1.80–1.74(m,1H),1.72–1.65(m,3H),1.62(t,J＝6.6Hz,5H),1.55–1.41(m,3H),1.33(dd,J＝14.9,6.7Hz,5H),1.27(s,9H),1.20–1.09(m,1H),1.08–1.02(m,1H),1.00(s,1H),0.98(d,J＝6.1Hz,3H),0.91–0.84(m,6H),0.81(d,J＝2.8Hz,4H),0.79(s,2H).13C NMR(101MHz,CDCl3)δ173.84,131.41,128.42,79.16,64.57,54.28,53.92,38.83,37.86,36.93,36.50,33.58,33.51,32.02,31.90,29.55,29.33,29.28,28.67,28.05,27.64,25.96,24.68,22.70,21.90,21.74,19.48,15.38,14.27,14.14.
Example 79
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-2-yl) heptyl ] propanamide
The first step compound 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (II) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing anhydrous DMF (N, N-dimethylformamide) (1 mL), HATU (81.34 mg,0.214mmol,1.2 eq) and DIEA (N, N-diisopropylethylamine) (0.059 mL, 0.317 mmol,2.0 eq) were added followed by stirring for 0.5hr, and 7- (pyridin-4-yl) hept-1-amine (V) (51.4 mg, 0.265 mmol,1.5 eq) was added followed by further stirring for 1hr. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a yellow oil. The crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 49:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (79-1) (80 mg,0.128mmol, 72.0%) as a yellow oil .1H NMR(400MHz,CDCl3)δ8.56(s,2H),7.43(s,2H),5.42(s,1H),3.27–3.15(m,3H),2.76(s,2H),2.37(m,2H),2.29–2.17(m,2H),2.07(m,5H),1.91(m,6H),1.81–1.75(m,2H),1.62(s,3H),1.59(m,7H),1.13–0.95(m,3H),0.92(s,1H),0.88(d,J＝5.8Hz,12H),0.83(d,J＝11.2Hz,2H),0.79(s,3H),0.75(s,3H),0.03(d,J＝2.8Hz,6H).
The second step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (80 mg,0.128mmol,1.0 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (1 mL), and after addition of a tetrahydrofuran solution of tetrabutylammonium fluoride (1.28 mL,1.28mmol,10 eq) was warmed to 50℃and stirred for 16hrs. TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil, which was extracted with ethyl acetate (10 mL x 3) and the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil. The crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0 to 20:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (79) (5.76 mg,0.010mmol, 7.55%) as a white solid .1H NMR(400MHz,CDCl3)δ8.52(s,2H),7.26–7.24(m,2H),5.45–5.32(m,1H),3.30–3.14(m,3H),2.68(t,J＝7.7Hz,2H),2.49–2.28(m,2H),2.28–2.09(m,3H),2.07–1.83(m,4H),1.76(d,J＝13.0Hz,2H),1.66(m,4H),1.62(s,3H),1.51–1.44(m,2H),1.34(s,8H),1.05(m,2H),0.97(s,3H),0.89(m,2H),0.80(d,J＝7.3Hz,6H).13C NMR(101MHz,DMSO)δ168.15,143.23,127.13,125.15,123.75,119.69,74.29,72.60,72.49,72.28,71.96,49.52,49.16,34.68,34.08,33.13,32.17,31.75,30.86,30.62,28.81,27.36,25.38,24.93,24.59,24.28,23.30,22.87,22.47,22.01,20.40,17.15,17.04,14.84,10.64,9.54.LC-MS(ESI)[M+H]+＝509.
Example 80
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-hydroxyethyl) propionamide
Reference example 27 conversion of (1-methylindol-5-yl) methane amine to diaminoethanol gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (2-hydroxyethyl) propionamide (80).1H NMR(400MHz,CDCl3)δ6.03(s,1H),3.73(t,J＝4.9Hz,2H),3.43 -3.42(m,2H),3.22(dd,J＝11.5,4.7Hz,1H),2.49–2.12(m,5H),2.09–1.85(m,5H),1.77(dt,J＝13.2,3.5Hz,1H),1.73–1.57(m,6H),1.37(ddd,J＝25.8,13.0,3.6Hz,1H),1.28–1.11(m,1H),1.06(dd,J＝12.3,4.9Hz,1H),1.03–0.95(m,3H),0.93-0.85(m,2H),0.83 0.79(m,6H).13C NMR(101MHz,CDCl3)δ174.48,131.62,128.73,79.05,62.69,54.28,53.93,42.58,38.83,37.93,36.93,36.51,35.42,33.55,32.13,28.04,27.62,25.11,21.88,21.82,19.54,15.34,14.27.LC-MS(ESI)[M+H]+＝378.
Example 81
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [ (3-methylphenyl) methyl ] propionamide
In the first step 3- ((4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthren-1-yl) propionic acid (III) (0.40 g,0.120mmol,1.0 eq) was added to a reaction flask containing DMF (1 mL), HATU (59.10 mg,0.156mmol,1.3 eq) and DIEA (N, N-diisopropylethylamine) (0.040 mL,0.240mmol,2.0 eq) were added followed by stirring for 0.5hr and stirring was continued at room temperature for 1hr after the addition of (3-methylphenyl) methanamine (18.84 mg,0.156mmol,1.3 eq). TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. Water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0-2:1), and the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ (3-methylphenyl) methyl ] propionamide (81) (8.87 mg, 92.1% purity, 31.2%) was obtained as a white solid .1H NMR(400MHz,CDCl3)δ7.22(t,J＝7.5Hz,1H),7.08(t,J＝9.9Hz,3H),5.71(s,1H),4.38(d,J＝5.3Hz,2H),3.21(dd,J＝11.5,4.6Hz,1H),2.55–2.41(m,1H),2.35(m,1H),2.33(s,3H),2.29(m,1H),2.24–2.11(m,2H),2.04–1.88(m,2H),1.86–1.64(m,5H),1.59(s,3H),1.43–1.24(m,2H),1.11–0.98(m,2H),0.96(s,3H),0.91(m,1H),0.85(m,2H),0.82–0.73(d,J＝2.5Hz,6H).).13C NMR(101MHz,CDCl3)δ172.68,138.46,138.21,131.73,128.72,128.65,128.31,124.95,79.06,77.33,77.22,77.02,76.70,54.28,53.91,43.75,38.82,37.80,36.92,36.49,35.38,33.53,32.11,28.04,27.62,25.05,21.89,21.70,21.37,19.55,15.36,14.27.LC-MS(ESI)[M+H]+＝438.
Example 82
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-4-yl) heptyl ] propionamide
Referring to the synthesis of intermediate V, 2-methylpyridine is exchanged for 4-methylpyridine to give 7- (pyridin-4-yl) hept-1-amine (82-1)
Reference example 27 conversion of (1-methylindol-5-yl) methanamine to 7- (pyridin-4-yl) hept-1-amine (82-1) gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- [7- (pyridin-4-yl) heptyl ] propanamide (82).1H NMR(400MHz,CDCl3)δ8.52(s,2H),7.26–7.24(m,2H),5.45–5.32(m,1H),3.30–3.14(m,3H),2.68(t,J＝7.7Hz,2H),2.49–2.28(m,2H),2.28–2.09(m,3H),2.07–1.83(m,4H),1.76(d,J＝13.0Hz,2H),1.66(m,4H),1.62(s,3H),1.51–1.44(m,2H),1.34(s,8H),1.05(m,2H),0.97(s,3H),0.89(m,2H),0.80(d,J＝7.3Hz,6H).13C NMR(101MHz,DMSO)δ168.15,143.23,127.13,125.15,123.75,119.69,74.29,72.60,49.52,49.16,34.68,34.08,33.13,32.17,31.75,30.86,30.62,28.81,27.36,25.38,24.93,24.59,24.28,23.30,22.87,22.47,22.01,20.40,17.15,17.04,14.84,10.64,9.54.LC-MS(ESI)[M+H]+＝509.
Example 83
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1- (4-methylpiperazin-1-yl) propan-1-one
Reference example 27 conversion of (1-methylindol-5-yl) methanamine to 1-methylpiperazine gives the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -1- (4-methylpiperazin-1-yl) propan-1-one (83).1H NMR(400MHz,CDCl3)δ3.67(m,2H),3.56–3.48(m,2H),3.22(dd,J＝11.4,4.6Hz,1H),2.44(m,4H),2.37(m,2H),2.35(s,3H),2.32–2.12(m,3H),2.01(m 2H),1.95–1.86(m,2H),1.76(m,1H),1.68(m,2H),1.62(s,3H),1.37(m,2H),1.06(m,2H),0.98(s,3H),0.95–0.85(m,3H),0.84–0.78(m,6H).13C NMR(101MHz,CDCl3)δ171.51,132.14,128.16,79.06,77.34,77.22,77.02,76.70,55.10,54.64,54.30,53.95,45.86,45.22,41.19,38.83,38.18,36.95,36.51,33.48,32.29,32.14,28.06,27.63,24.87,21.92,21.77,19.55,15.38,14.30.LC-MS(ESI)[M+H]+＝417.2.
Example 84
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (1, 4-oxaazepin-4-yl) hexyl ] propanamide
In the first step [ (6-bromohexyl) amino ] methanoic acid-2-methylpropan-2-yl ester (200 mg, 0.514 mmol,1.0 eq) and morpholine (0.070 mL,1.07mmol,1.5 eq) were added to a reaction flask containing DMF (N, N-dimethylformamide) (5 mL), cesium carbonate (581 mg,1.78mmol,2.5 eq) was added, and the mixture was warmed to 100℃and stirred for 6hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), affording a colorless oil. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 1:3) to give { [6- (1, 4-oxazacyclohex-4-yl) hexyl ] amino } methanoic acid-2-methylpropan-2-yl ester (84-1) (140 mg, purity 100%, 68.4%) as a colorless oil. 1 H NMR (400 mhz, dmso) delta 3.56 (m, 4H), 2.88 (dd, j=13.0, 6.5hz, 2H), 2.28 (m, 6H), 1.35 (m, 12H), 1.24 (m, 4H).
In the second step { [6- (1, 4-oxazacyclohex-4-yl) hexyl ] amino } methanoic acid-2-methylpropan-2-yl ester (84-1) (70 mg,0.244mmol,1.0 eq) was added to a solution of hydrogen chloride (2 mL,8.00mmol,32 eq) in methanol (2 mL) and stirred overnight at room temperature. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a colorless oil, which was used directly in the subsequent reaction without further purification. Crude 6- (1, 4-oxazacyclohex-4-yl) hex-1-amine (84-2) (50 mg, 100% purity, 91.8%) was obtained as a colorless oil .1H NMR(400MHz,DMSO)δ11.33(s,1H),8.03(s,3H),3.90–3.77(m,4H),3.39-3.36(d,J＝12.3Hz,2H),3.06–2.97(m,4H),2.78-2.73(dd,J＝13.6,6.4Hz,2H),1.73–1.69(m,2H),1.59–1.55(m,2H),1.36–1.31(m,4H).
In a third step 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.178mmol,1.0 eq) was added to a reaction flask containing DMF (N, N-dimethylformamide) (2 mL), HATU (88.12 mg,0.232mmol,1.3 eq) and DIEA (N, N-diisopropylethylamine) (0.059 mL, 0.351 mmol,2.0 eq) were added and stirred for 0.5hr at room temperature after addition of 6- (1, 4-oxaazepin-4-yl) hex-1-amine (84-2) (43.1 mg,0.232mmol,1.3 eq). TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. Water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil. The crude product was isolated and purified by column chromatography (dichloromethane: methanol=1:0-16:1) to give the product 3- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (1, 4-oxaazepin-4-yl) hexyl ] propanamide (84-3) (40 mg, purity 100%, 36.3%) as a pale yellow solid .1H NMR(400MHz,CDCl3)δ5.51(s,1H),3.88(s,4H),3.21(m,,3H),2.66(m,5H),2.47–2.19(m,3H),2.18–1.65(m,14H),1.63(s,3H),1.61–1.43(m,6H),1.31(m,1H),1.11–0.90(m,4H),0.89(s,9H),0.86(m,3H),0.81(s,3H),0.75(s,3H),0.03(d,J＝2.6Hz,6H).
In the fourth step, 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (1, 4-oxaazepin-4-yl) hexyl ] propanamide (84-3) (40 mg,0.065mmol,1.0 eq) was dissolved in a tetrahydrofuran solution (1 mol/L,5 mL) of tetrabutylammonium fluoride and the reaction solution was stirred at 50℃for 16hrs. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃) to give a yellow oil, which was extracted with ethyl acetate (10 mL x 3) and the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a colorless oil. The crude product was purified by column chromatography (dichloromethane: methanol=1:0-16:1) and preparative HPLC to give the product 3- [ (4 br,7 s) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [6- (1, 4-oxaazepin-4-yl) hexyl ] propionamide (84) (11.7 mg, purity 98.0%, 35.3%) as a white solid .1H NMR(400MHz,CDCl3)δ5.51(s,1H),3.79(t,J＝4.4Hz,4H),3.31–3.16(m,3H),2.58(m,4H),2.49–2.41(m,3H),2.40–2.07(m,5H),2.03–1.87(m,3H),1.84–1.66(m,4H),1.63(s,3H),1.59(m,2H),1.49(m,2H),1.34(s,5H),1.16–1.01(m,2H),0.97(s,3H),0.94–0.84(m,4H),0.80(d,J＝8.8Hz,6H).13C NMR(101MHz,CDCl3)δ173.03,131.87,128.45,79.05,77.33,77.22,77.01,76.70,65.33,58.16,54.26,53.91,52.62,39.24,38.81,37.90,36.91,36.49,35.59,33.54,32.09,29.39,28.04,27.60,26.62,26.37,25.18,24.61,21.89,21.78,19.58,15.37,14.29.LC-MS(ESI)[M+H]+＝503.3.
Example 85
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [ (4-methylphenyl) methyl ] propionamide
Referring to example 81, (3-methylphenyl) methanamine was changed to (4-methylphenyl) methanamine to give the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [ (4-methylphenyl) methyl ] propanamide (85).1H NMR(400MHz,CDCl3)δ7.19–7.12(m,4H),5.69(s,1H),4.38(d,J＝5.3Hz,2H),3.21(dd,J＝11.5,4.6Hz,1H),2.53–2.36(m,2H),2.34(s,3H),2.31–2.24(m,1H),2.23–2.11(m,2H),1.95(m,2H),1.87–1.64(m,5H),1.59(s,3H),1.48–1.27(m,2H),1.08–0.98(m,2H),0.96(s,3H),0.90(m,1H),0.86–0.81(m,2H),0.79(d,J＝3.5Hz,6H).13C NMR(101MHz,CDCl3)δ172.71,137.51,135.28,131.74,129.39,128.64,127.89,79.06,77.33,77.21,77.01,76.69,54.28,53.91,43.48,38.82,37.81,36.92,36.49,35.43,33.53,32.10,28.03,27.62,25.06,21.88,21.70,21.09,19.55,15.35,14.26.LC-MS(ESI)[M+H]+＝438.
Example 86
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (9-methoxynonyl) propionamide
In the first step, 9-bromononon-1-ol (2.0 g,9.0mmol,1.0 eq) was dissolved in methanol (40 mL), sodium methoxide (4.84 g,89.6mmol,10.0 eq) was added, and the reaction solution was stirred at 80℃for 2h. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction solution was concentrated, and the crude product was petroleum ether: ethyl acetate=5:1, and concentrated to give 9-methoxynonen-1-ol (86-1) (1.1 g, purity 90%, yield 63.38％).1H NMR(400MHz,CDCl3)δ3.56(t,J＝6.7Hz,2H),3.29(t,J＝6.7Hz,2H),3.26(s,3H),1.64(s,1H),1.54–1.44(m,4H),1.24(s,9H).
In a second step 9-methoxynonen-1-ol (86-1) (0.50 g,2.87mmol,1.0 eq) was dissolved in acetonitrile (10 mL) followed by carbon tetrabromide (1.43 g,4.30mmol,1.5 eq) and triphenylphosphine (1.13 g,4.30mmol,1.5 eq) and the reaction stirred at room temperature for 16hrs, TLC (petroleum ether: ethyl acetate=10:1) indicated that the reaction was complete. After concentration, the crude product was purified with petroleum ether: ethyl acetate=20:1 was passed through a column to give 1-bromo-9-methoxynonane (86-2) (0.6 g, purity 90%, yield 79.4％).1H NMR(400MHz,CDCl3)δ3.41(t,J＝6.9Hz,2H),3.36(t,J＝6.6Hz,2H),3.33(s,3H),1.92–1.79(m,1H),1.62–1.51(m,1H),1.42(t,J＝6.9Hz,1H),1.31(s,4H).
In the third step 9-bromo-1-methoxynonane (85-2) (0.6 g,2.53mmol,1.0 eq) and potassium phthalimide (0.57 g,3.04mmol,1.2 eq) were dissolved in N, N-dimethylformamide (10 mL), the reaction stirred overnight at room temperature and TLC (petroleum ether: ethyl acetate=5:1) showed completion of the reaction. After the reaction, the mixture was extracted with ethyl acetate (100 mL), washed with water (50 mL. Times.5), dried over anhydrous Na 2SO4, and the crude product was purified with petroleum ether: ethyl acetate=20:1-5:1 was passed through a column to give 2- (9-methoxynonyl) isoindole-1, 3-dione (86-3) (0.56 g, purity 90%, yield 65.4%). LC-MS (ESI) [ m+h ] + = 304.14.
In the fourth step, 2- (9-methoxynonyl) isoindole-1, 3-dione (86-3) (0.6 g,1.98mmol,1.0 eq) was dissolved in ethanol (20 mL), hydrazine hydrate (0.48 mL,9.89mmol,5.0 eq) was added, the reaction was stirred at 80℃for 2h, and TLC (petroleum ether: ethyl acetate=1:1) showed completion. After the completion of the reaction, the mixture was concentrated, extracted with chloroform (20 mL), washed with water (20 mL. Times.2), and dried over anhydrous Na 2SO4 to give a crude 9-methoxynon-1-amine (86-4) which was directly fed to the next step.
In the fifth step, 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (I) (80 mg,0.18mmol,1.0 eq), HATU (81.3 mg,0.21mmol,1.2 eq) and N, N-diisopropylethylamine (29.9 mg,0.23mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3 mL), and after stirring for 2min the reaction mixture was added 9-methoxynon-1-amine (86-4) (37.7 mg,0.2mmol,1.2 eq) and stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=3:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 5:1 passage through a column afforded 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (9-methoxynonyl) propanamide (86-5) (50 mg, 90% purity, 41.8% yield).
In the sixth step, 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (9-methoxynonyl) propanamide (86-5) (50 mg,0.08mmol,1.0 eq) was dissolved in THF (tetrahydrofuran) (1 mol/L,3 mL) of TBAF, and the reaction solution was stirred at 50℃for 3hrs. TLC (petroleum ether: ethyl acetate=3:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=3:1, and concentrated to give 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (9-methoxynonyl) propionamide (86) (2.33 mg, 96.84% purity, yield) as a colorless oil 5.7％).1H NMR(400MHz,CDCl3)δ5.46(s,1H),3.36(t,J＝6.6Hz,2H),3.33(s,3H),3.22(dd,J＝11.4,4.6Hz,3H),2.48–2.39(m,1H),2.36-2.34(s,1H),2.29–2.20(m,1H),2.19–2.10(m,2H),2.05-1.90(m,3H),1.76(dd,J＝13.2,3.4Hz,1H),1.73–1.67(m,3H),1.70-1.765(m,3H),1.63(s,3H),1.58(d,J＝5.7Hz,1H),1.55(d,J＝6.9Hz,1H),1.50-1.47(m,2H),1.35-1.32(m,1H),1.30-1.27(m,7H),1.25(s,1H),1.07–0.99(m,2H),0.97(s,3H),0.89–0.83(m,2H),0.80(d,J＝9.2Hz,6H).13C NMR(101MHz,CDCl3)δ173.29,131.89,128.52,79.07,77.35,77.24,77.03,76.71,72.94,58.57,54.28,53.93,39.55,38.83,37.88,36.93,36.51,35.60,33.57,32.11,29.67,29.64,29.47,29.42,29.24,28.05,27.62,26.92,26.12,25.15,21.90,21.79,19.60,15.38,14.30.LC-MS(ESI)[M+H]+＝490.40.
Example 87
Preparation of the Compound N- [ (4-fluorophenyl) methyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide
Referring to example 81, (3-methylphenyl) methanamine was converted to (4-fluorophenyl) methanamine to give the compound N- [ (4-fluorophenyl) methyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (87).1H NMR(400MHz,CDCl3)δ7.26–7.20(t,J＝8.6Hz,2H),7.03-6.99(t,J＝8.6Hz,2H),5.78(s,1H),4.46–4.33(m,2H),3.22-3.19(dd,J＝11.5,4.6Hz,1H),2.52–2.43(m,1H),2.32-2.27(m,2H),2.24–2.10(m,2H),2.02–1.89(m,2H),1.88–1.65(m,5H),1.59(s,3H),1.48–1.20(m,2H),1.07–0.98(m,2H),0.95(s,3H),0.93–0.81(m,3H),0.79(d,J＝3.0Hz,6H).13C NMR(101MHz,CDCl3)δ172.75,163.42,160.98,134.19,131.68,129.57,129.49,128.75,115.65,115.44,79.04,77.33,77.02,76.70,54.28,53.90,42.94,38.82,37.80,36.91,36.49,35.36,33.55,32.10,28.04,27.62,25.02,21.89,21.71,19.56,15.36,14.26.LC-MS(ESI)[M+H]+＝442.
Example 88
Preparation of methyl Compound 9- ({ 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) nonanoate
In the first step, DMP (dess-Martin reagent) (2.93 g,6.9mmol,2.0 eq) was dispersed in methylene chloride (methylene chloride) (5 mL), and a methylene chloride solution (10 mL) of 2- (9-hydroxynonyl) isoindole-1, 3-dione (70-1) (1.0 g,3.5mmol,1.0 eq) was added under ice-bath, and the reaction solution was stirred at room temperature for 1hrs. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. The reaction mixture was diluted with dichloromethane (50 mL) and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=15:1, and concentrating to obtain 9- (1, 3-dioxy-2, 3-dihydro-1H-isoindol-2-yl) nonanal (88-1), which is directly taken into the next step.
In a second step 9- (1, 3-dioxy-2, 3-dihydro-1H-isoindol-2-yl) nonanal (88-1) (0.20 g,0.70mmol,1.0 eq) was dissolved in t-BuOH (t-butanol) (8 mL), and 2-methyl-2-butene (0.70 g,3.5mmol,5.0 eq) and an aqueous solution (5 mL) of sodium dihydrogen phosphate (0.25 g,2.1mmol,3.0 eq) and sodium chlorite (0.19 g,2.1mmol,3.5 eq) were added under ice bath and the reaction solution was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=5:1, uv254nm color development) showed the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (20 mL), and washed with saturated sodium bicarbonate (20 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=3:1, and concentrated to give a colorless semi-solid, which was directly taken to the next step.
Crude 9- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) nonanoic acid (0.20 g,0.66mmol,1.0 eq) was dissolved in methanol (10 mL), concentrated sulfuric acid (1 mL) was added, the reaction was allowed to proceed for 5min at room temperature, and TLC (Petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. After the reaction solution is concentrated, petroleum ether is used as a crude product: ethyl acetate=5:1, and concentrating to give methyl 9- (1, 3-dioxy-2, 3-dihydro-1H-isoindol-2-yl) nonanoate (88-2) (0.19 g, 90% purity, two-step yield 85.5％).1H NMR(400MHz,CDCl3)δ7.76(dt,J＝7.0,3.5Hz,2H),7.66–7.62(m,2H),3.65–3.56(m,5H),2.22(t,J＝7.5Hz,2H),1.62–1.49(m,4H),1.27–1.15(m,8H).
In a third step methyl 9- (1, 3-dioxo-2, 3-dihydro-1H-isoindol-2-yl) nonanoate (88-2) (0.19 g,0.60mmol,1.0 eq) was dissolved in EtOH (ethanol) (10 mL) and hydrazine hydrate (0.15 g,3.0mmol,5.0 eq) was added and reacted at 80℃for 3H, TLC (Petroleum ether: ethyl acetate=1:1) indicated that the reaction was complete. After the reaction solution was concentrated, it was extracted with chloroform (20 mL), washed with water (20 mL), and the crude product was concentrated on an organic layer using petroleum ether: ethyl acetate=2:1 was passed through a column to give methyl 9-aminononanoate (88-3) (0.13 g, purity 80%, two step yield 46.67％).1H NMR(400MHz,MeOD)δ3.55(d,J＝2.5Hz,3H),2.51(dd,J＝9.7,4.6Hz,2H),2.21(t,J＝7.4Hz,2H),1.51(d,J＝6.7Hz,2H),1.37(s,2H),1.23(s,8H).
In the fourth step, 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanoic acid (III) (140 mg,0.31mmol,1.0 eq), HATU (142 mg,0.37mmol,1.2 eq) and N, N-diisopropylethylamine (52.4 mg,0.40mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (4 mL), and after stirring for 2min, the reaction mixture was added to methyl 9-aminononanoate (88-3) (70 mg,0.37mmol,1.2 eq) and stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1-5:1 was passed through a column to give methyl 9- ({ 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) nonanoate (88-3) (90 mg, purity 90%, yield 42.01％).1H NMR(400MHz,CDCl3)δ3.67(s,3H),3.31–3.13(m,3H),2.46(s,1H),2.30(t,J＝7.4Hz,3H),2.17-2.14(m,2H),2.00-1.80(m,6H),1.74–1.43(m,13H),1.30-1.26(m,9H),0.94–0.85(m,13H),0.85–0.71(m,7H),0.03(d,J＝2.7Hz,6H).
In the fifth step, methyl 9- ({ 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) nonanoate (88-3) (80 mg,0.13mmol,1.0 eq) was dissolved in THF (tetrahydrofuran) (1 mol/L,5 mL) of TBAF, and the reaction solution was stirred at 80℃for 5hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1, and concentrated to give 9- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) nonanoic acid (88) (18 mg, 90% purity, yield as a colorless oil 25.0％).1H NMR(400MHz,DMSO)δ11.98(s,1H),7.71(s,1H),4.33(s,1H),3.02-2.96(m,3H),2.35–2.24(m,1H),2.18(t,J＝7.4Hz,3H),2.09–2.04(m,1H),2.01–1.89(m,3H),1.83(d,J＝12.9Hz,1H),1.71–1.59(m,3H),1.55(s,2H),1.52-1.45(m,4H),1.40-1.30(m,3H),1.24(s,10H),1.04–0.95(m,2H),0.91–0.85(m,4H),0.81–0.75(m,5H),0.72-0.69(m,3H).13C NMR(101MHz,DMSO)δ175.01,172.10,132.76,127.12,77.38,54.37,53.97,40.60,40.39,40.18,39.97,39.77,39.56,39.35,38.93,38.83,37.84,37.04,36.55,35.17,34.17,33.55,32.16,29.64,29.50,29.18,29.13,29.00,28.65,27.90,26.83,25.52,24.98,22.04,21.86,19.77,16.28,14.57.LC-MS(ESI)[M+H]+＝490.
Example 89
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (pyridin-4-ylmethyl) propionamide
Referring to example 81, (3-methylphenyl) methanamine was changed to pyridin-4-ylmethylamine to give the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (pyridin-4-ylmethyl) propanamide (89).1H NMR(400MHz,CDCl3)δ8.55(d,J＝4.4Hz,2H),7.26–7.23(m,2H),6.03(s,1H),4.48(t,J＝6.7Hz,2H),3.22(dd,J＝11.5,4.5Hz,1H),2.52-2.50(m,1H),2.43–2.31(m,2H),2.29–2.13(m,3H),2.02–1.85(m,3H),1.79–1.55(m,8H),1.47–1.02(m,3H),1.03–0.92(m,4H),0.90–0.83(m,2H),0.81-0.79(m,5H).13C NMR(101MHz,CDCl3)δ173.20,148.95,131.53,128.87,122.66,79.02,77.35,77.23,77.03,76.71,54.26,53.99,53.88,42.41,38.83,37.83,36.90,36.50,35.33,33.56,32.12,28.05,27.61,25.04,21.90,21.75,21.42,19.60,15.39,14.29.LC-MS(ESI)[M+H]+＝425.0.
Example 90
Preparation of the Compound N- (9-aminononyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide hydrochloride
In the first step, nono-1, 9-diamine (100 mg,0.63mmol,1.0 eq) was dissolved in methylene chloride (methylene chloride) (5 mL), and a solution of triethylamine (0.18 mL,1.26mmol,5.0 eq), (BOC) 2 O (0.06 mL,0.25mmol,0.4 eq) in methylene chloride (5 mL) was added and the reaction mixture was stirred at room temperature for 16h. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. The reaction mixture was diluted with methylene chloride (50 mL) and washed with saturated ammonium chloride (40 mL. Times.2). The organic layer was dried over anhydrous sodium sulfate, and the crude product was purified by passing through a column with dichloromethane: methanol=10:1-5:1, and concentrated to give [ (9-aminononyl) amino ] methanoic acid-2-methylpropan-2-yl ester (90-1) (50 mg, purity 50%, yield) as a colorless liquid 38.2％).1H NMR(400MHz,DMSO)δ7.87(s,2H),3.32–3.27(m,2H),2.77–2.71(m,2H),1.60–1.46(m,2H),1.44–1.30(m,10H),1.24(s,11H).
In the second step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (40 mg,0.12mmol,1.0 eq), HATU (54.5 mg,0.14mmol,1.2 eq) and N, N-diisopropylethylamine (20.1 mg,0.15mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (1.5 mL), and after stirring for 2min the reaction [ (9-aminononyl) amino ] methanoic acid-2-methylpropan-2-yl ester (90-1) (40 mg,0.15mmol,1.3 eq) was added and the reaction mixture was stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=3:1) showed that the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 3:1 column passed to give { [9- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) nonyl ] amino } methanesulfonic acid-2-methylpropan-2-yl ester (90-2) (30 mg, purity 90%, yield 39.3%) the product was directly taken to the next step.
In the third step, 2-methylpropan-2-yl { [9- ({ 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) nonyl ] amino } methanoate (90-2) (30 mg,0.05mmol,1.0 eq) was dissolved in (1 mol/L,2 mL), and the reaction mixture was stirred at room temperature for 2hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The crude product after concentration of the reaction solution was subjected to column chromatography with methylene chloride, methanol=50:1-10:1, and concentrated to give white solid N- (9-aminononyl) -3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (90) (35 mg, purity 90%, yield 35.3％).1H NMR(400MHz,DMSO)δ7.97(s,3H),7.77(s,1H),4.34(d,J＝4.9Hz,1H),3.06–2.89(m,3H),2.73(s,2H),2.51(s,9H),2.32–1.80(m,8H),1.68–1.43(m,11H),1.38–1.21(m,16H),1.07–0.86(m,6H),0.80–0.74(m,5H),0.71-0.69(m,3H).13C NMR(101MHz,DMSO)δ172.14,132.78,127.11,77.36,54.37,53.97,40.63,40.42,40.21,40.00,39.79,39.59,39.38,39.15,38.92,38.84,37.86,37.04,36.54,35.18,33.54,32.15,29.66,29.26,29.13,28.98,28.66,27.89,27.39,26.86,26.32,25.52,22.05,21.85,19.78,16.29,14.58.
Example 91
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (3-cyano-1-methylindol-5-yl) amino ] methanamide
In the first step, the compound 1-methyl-5-nitroindole-3-carbaldehyde (100 mg,0.490mmol,1.0 eq), hydroxylamine hydrochloride (68.06 mg,0.979mmol,2.0 eq), sodium formate (133.23 mg,1.959mmol,4.0 eq) were dissolved in formic acid (1.3 mL) and N-methylpyrrolidone (0.13 mL), then heated to 105℃and stirred for 3h, LC-MS detection, after completion of the reaction, cooled to room temperature, then water was added, and the precipitated solid was filtered and dried to give 1-methyl-5-nitroindole-3-carbonitrile (91-1) (98 mg,0.487mmol, yield: 99.46%) as a yellow solid compound. LC-MS (ESI) [ m+na ] + =224.1
In the second step, the compound 1-methyl-5-nitroindole-3-carbonitrile (91-1) (50 mg,0.249mmol,1.0 eq) was dissolved in ethanol (5 mL), then ammonium chloride (132.93 mg, 2.480 mmol,10.0 eq) and zinc powder (259.97 mg,3.976mmol,16.0 eq) were added thereto, the temperature was raised to 50 degrees and stirred for 1 hour, after completion of the reaction, filtration and then spin-drying gave a crude product, which was separated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give 5-amino-1-methylindole-3-carbonitrile (91-2) (36 mg,0.21mmol, yield: 84.21%) as an oily compound. LC-MS (ESI) [ m+na ] + =172.1
The third step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing toluene (2 mL), diphenyl azide phosphate (64.39 mg,0.234mmol,1.05 eq) and triethylamine (0.065 mL, 0.268 mmol,2.0 eq) were added and stirred at room temperature (90 ℃) for 1hr, the reaction mixture was cooled to room temperature, tetrahydrofuran (2 mL) was further added, and stirring was continued for 18hr with 5-amino-1-methylindole-3-carbonitrile (91-2) (45.81 mg,0.223mmol,1.0 eq). LC-MS monitoring was incomplete. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } -1- [ (3-cyano-1-methylindol-5-yl) amino ] methanamide (91-3) (330 mg,0.045mmol, 20.39%) as a pale yellow solid. LC-MS (ESI) [ m+h ] + = 618.40
In the fourth step N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopentan-5-yl) amino ] methanamide (91-3) (30 mg,0.049mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.4816 mL, 0.4816 mmol,10.0 eq) was added thereto and then heated to 50℃for 18hr with stirring. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by preparative TLC (dichloromethane: methanol=10:1), to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (3-cyano-1-methylindol-5-yl) amino ] methanamide (91) (10 mg,0.020mmol, 40.15%) as a white solid .1H NMR(400MHz,CDCl3)δ7.85(d,J＝2.2Hz,1H),7.72(d,J＝1.8Hz,1H),7.44(d,J＝8.8Hz,1H),7.27(dd,J＝8.8,2.0Hz,1H),3.85(s,3H),3.26(dd,J＝8.6,5.0Hz,1H),3.14(d,J＝10.0Hz,2H),2.38(m,1H),2.29(m,2H),2.00(d,J＝21.5Hz,2H),1.91(dd,J＝16.3,4.6Hz,1H),1.76(s,3H),1.66(s,3H),1.62(d,J＝1.8Hz,2H),1.40(s,2H),1.33(s,1H),1.28(s,3H),1.08(m,2H),0.96(d,J＝5.1Hz,3H),0.90(d,J＝6.0Hz,1H),0.86(d,J＝5.5Hz,3H),0.79(d,J＝8.1Hz,3H).13C NMR(101MHz,CDCl3)δ157.37,136.79,134.56,132.76,130.10,129.07,128.14,117.48,115.62,110.69,109.22,83.54,78.37,54.43,54.20,38.52,37.92,36.90,36.22,33.26,32.38,32.17,29.34,29.12,27.24,26.93,21.69,21.58,18.52,14.72,13.40.LC-MS(ESI)[M+H]+＝504.30
Example 92
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } undecanamide
Undecanoic acid (42.6 mg,0.23mmol,1.4 eq), HATU (74.6 mg,0.20mmol,1.2 eq) and N, N-diisopropylethylamine (27.5 mg,0.21mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (1.5 mL), and after stirring for 2min the reaction (4 ar,4 bs) -8- (2-aminoethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (65-2) (50 mg,0.16mmol,1.0 eq) was added and the reaction stirred at room temperature for 20min. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate = 10:1 column chromatography gave N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } undecanamide (92) (3.66 mg, 91% purity, yield) 4.4％).1H NMR(400MHz,CDCl3)δ5.51(s,1H),3.38–3.29(m,1H),3.22(dd,J＝11.5,4.6Hz,1H),3.15–3.03(m,1H),2.28(t,J＝7.3Hz,2H),2.19–2.12(m,2H),2.06–1.89(m,3H),1.78–1.58(m,11H),1.27(d,J＝9.7Hz,15H),1.17–0.95(m,6H),0.93–0.83(m,6H),0.81(s,3H),0.79(s,2H).13C NMR(101MHz,CDCl3)δ171.64,83.37,79.31,77.34,77.23,77.02,76.71,55.26,54.18,46.44,39.41,39.22,37.52,37.12,36.41,31.60,28.70,28.56,27.89,25.91,21.46,21.32,20.09,19.81,18.11,16.00,14.24,-3.79,-4.92.LC-MS(ESI)[M+H]+＝474.
Example 93
Preparation of Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [1- (7-phenylheptyl) -1,2, 3-triazacyclopentan-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
The first step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (100 g,0.231mmol,1.0 eq) was added to a reaction flask containing methanol (5 mL), dimethyl (1-diazonium-2-oxosubunit propyl) phosphonate (66.59 mg,0.347mmol,1.5 eq) and potassium carbonate (66.85 mg, 0.4632 mmol,2 eq) were added and reacted at room temperature for 1 hour. TLC (petroleum ether: ethyl acetate=20:1) monitored reaction completion. Dichloromethane (30 mL) was added to the reaction solution, which was washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=100:1) to give the product { [ (2 s,4ar,4bs,8ar,10 ar) -8- (but-3-ynyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecaphenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (93-1) (80 mg,0.187mmol, 80.74%) as a white solid .1H NMR(400MHz,CDCl3)δ3.15(dd,J＝11.2,4.6Hz,1H),2.34–2.14(m,3H),2.09(s,2H),1.90(dd,J＝5.5,2.9Hz,3H),1.86–1.74(m,1H),1.72–1.61(m,3H),1.59(s,3H),1.54(d,J＝18.0Hz,3H),1.34–1.27(m,1H),1.00(dd,J＝12.5,4.6Hz,1H),0.97–0.88(m,2H),0.85(d,J＝4.2Hz,12H),0.80(dd,J＝6.2,4.6Hz,1H),0.77(s,3H),0.73(d,J＝7.1Hz,3H),0.00(d,J＝2.6Hz,6H).
The second step compound bromobenzene (1 g,6.369mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (10 mL), hept-6-yn-1-ol (1.07 g,9.554mmol,1.5 eq), diphenylphosphine palladium dichloride (0.45 g,0.637mmol,0.1 eq), cuprous iodide (0.2 g,0.637mmol,0.1 eq), N, N-diisopropylethylamine (2.06 g,15.92mmol,2.5 eq) was added and stirred at 80℃for 2hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. Ethyl acetate (20 mL) was added to the reaction solution, which was washed with water (20 mL x 2), dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1), and the product 7-phenylhept-6-yn-1-ol (93-2) (800 mg,4.249mmol, 66.72%) was collected as a colorless oily liquid. LC-MS (ESI) [ m+h ] + =175.3
The third step compound, 7-phenylhept-6-yn-1-ol (93-2) (800 mg,4.249mmol,1.0 eq), was added to a reaction flask containing methanol (10 mL), palladium on carbon (160 mg) was added to replace hydrogen, pressurized to 40psi, warmed to 50℃and reacted for 18 hours. LC-MS monitored the reaction to completion, celite filtration, and spin-drying of the filtrate under reduced pressure (water pump, 40 ℃ C.) gave the product 6-phenylhex-1-ol (93-3) (750 mg,4.207mmol, 99%) as a colorless oil.
The fourth step compound 6-phenylhex-1-ol (93-3) (100 mg,0.520mmol,1.0 eq) was added to a reaction flask containing dichloromethane (5 mL), 4-methylbenzenesulfonyl chloride (297.41 mg,1.560mmol,3.0 eq), 4- (dimethylamino) pyridine (6.35 mg,0.052mmol,0.1 eq), ethyl [ di (propan-2-yl) ] amine (336.06 mg,2.60mmol,5 eq) was added and reacted at room temperature (25 degrees) for 18 hours. TLC (petroleum ether: ethyl acetate=20:1) monitored the reaction. Ethyl acetate (20 mL) was added to the reaction solution, which was washed with water (20 mL x 2), dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=50:1), and the product 4-methylbenzenesulfonic acid-7-phenylheptyl ester (93-4) (100 mg,0.289mmol, 55.50%) was collected as a white solid. LC-MS (ESI) [ m+h ] + =175.2
In the fifth step, 4-methylbenzenesulfonic acid-7-phenylheptyl ester (93-4) (100 mg,0.289mmol,1.0 eq) was dissolved in dimethyl sulfoxide (3 mL), then sodium azide (37.52 mg,0.577mmol,2.0 eq) was added thereto, nitrogen was replaced, then heated to 50 degrees and stirred overnight, and TLC (petroleum ether: ethyl acetate=20:1) monitored the reaction. After completion of the reaction, water was added thereto for quenching, followed by extraction with ethyl acetate (50 mL), and the organic phase was washed twice with saturated brine (50 mL x 2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give (7-azidoheptyl) benzene (93-5) (60 mg,0.276mmol, 95.66%) as an oily compound, and the crude product was directly used in the next step.
In the sixth step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (but-3-ynyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrofen-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (93-1) (30 mg,0.070mmol,1.0 eq), (7-azidoheptyl) benzene (93-5) (15 mg,0.070mmol,1 eq), sodium ascorbate (13.86 mg,0.070mmol,1 eq), copper pentahydrate (17.47 mg,0.070mmol,1 eq) were dissolved in t-butanol (1 mL) and H 2 O (1 mL), and then stirred overnight at room temperature (petroleum ether: ethyl acetate=20:1) and, after the reaction was completed, ethyl acetate was added to dilute it, and the crude product (50 mg,0.070mmol,1 eq) was obtained by washing with dry TLC (5, 7-azidoheptyl) benzene (93-5) (15 mg,0.070mmol,1 eq), sodium ascorbate (13.86 mg, 0.47 mmol, 1mg, 7-tetramethyl), copper sulfate (17.47 mmol,1 mL), and H 2 O (1 mL), and dry ethyl ether (1 mL) were used to extract dry off.
The seventh step was to dissolve the compound 4- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- (7-phenylheptyl) -1,2, 3-triazacyclopenta-ne (93-6) (25 mg,0.039mmol,1 eq) in tetrahydrofuran (0.5 mL), then add tetrabutylammonium fluoride (1M in tetrahydrofuran) (1.5 mL), displace nitrogen, warm up to 50℃and stir overnight. LC-MS detection reaction, when the reaction was completed, water was added thereto, then extraction was performed with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was then separated and purified (petroleum ether: ethyl acetate=2:1) via a preparative plate to obtain (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [1- (7-phenylheptyl) -1,2, 3-triazacyclopent-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (93) (19 mg,0.036mmol, yield: 92.33%) as a solid compound, which was freeze-dried to obtain a powdery compound .1H NMR(400MHz,CDCl3)δ7.33–7.26(m,3H),7.20(t,J＝7.7Hz,3H),4.32(t,J＝7.2Hz,2H),3.26(dd,J＝11.5,4.6Hz,1H),2.92–2.75(m,1H),2.71–2.60(m,3H),2.50–2.36(m,2H),2.26(dt,J＝9.1,4.8Hz,1H),2.09–1.98(m,2H),1.90(dt,J＝13.9,7.1Hz,4H),1.83–1.74(m,4H),1.67(ddd,J＝15.3,8.7,5.9Hz,5H),1.60(s,3H),1.47–1.38(m,2H),1.32(dd,J＝10.2,6.7Hz,3H),1.13–1.03(m,2H),1.01(d,J＝5.0Hz,3H),0.97–0.86(m,3H),0.84(d,J＝3.8Hz,3H),0.82(s,3H).13C NMR(101MHz,CDCl3)δ148.08,142.62,132.20,128.38,128.26,128.08,125.65,120.28,79.09,54.31,53.95,50.19,38.84,37.99,36.97,36.53,35.87,33.52,32.18,31.31,30.37,29.10,28.99,28.89,28.06,27.65,26.43,24.69,21.94,21.85,19.59,15.38,14.30.LC-MS(ESI)[M+1]+＝532.4.
Example 94
Preparation of Compound 9- ({ 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) nonanoic acid
In the first step, 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanoic acid (I) (140 mg,0.31mmol,1.0 eq), HATU (142 mg,0.37mmol,1.2 eq) and N, N-diisopropylethylamine (52.4 mg,0.40mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (4 mL), and after stirring for 2min, the reaction mixture was added to methyl 9-aminononanoate (88-3) (70 mg,0.37mmol,1.2 eq) and stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1-5:1 was passed through a column to give methyl 9- ({ 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) nonanoate (94-1) (90 mg, purity 90%, yield 42.01％).1H NMR(400MHz,CDCl3)δ3.67(s,3H),3.31–3.13(m,3H),2.46(s,1H),2.30(t,J＝7.4Hz,3H),2.17-2.14(m,2H),2.00-1.80(m,6H),1.74–1.43(m,13H),1.30-1.26(m,9H),0.94–0.85(m,13H),0.85–0.71(m,7H),0.03(d,J＝2.7Hz,6H).
In the fifth step, methyl 9- ({ 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionyl } amino) nonanoate (94-1) (80 mg,0.13mmol,1.0 eq) was dissolved in THF (tetrahydrofuran) (1 mol/L,5 mL) of TBAF, and the reaction solution was stirred at 80℃for 5hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1, and concentrated to give 9- ({ 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionyl } amino) nonanoic acid (94) (18 mg, 90% purity, yield as a colorless oil 25.0％).1H NMR(400MHz,DMSO)δ11.98(s,1H),7.71(s,1H),4.33(s,1H),3.02-2.96(m,3H),2.35–2.24(m,1H),2.18(t,J＝7.4Hz,3H),2.09–2.04(m,1H),2.01–1.89(m,3H),1.83(d,J＝12.9Hz,1H),1.71–1.59(m,3H),1.55(s,2H),1.52-1.45(m,4H),1.40-1.30(m,3H),1.24(s,10H),1.04–0.95(m,2H),0.91–0.85(m,4H),0.81–0.75(m,5H),0.72-0.69(m,3H).13C NMR(101MHz,DMSO)δ175.01,172.10,132.76,127.12,77.38,54.37,53.97,40.60,40.39,40.18,39.97,39.77,39.56,39.35,38.93,38.83,37.84,37.04,36.55,35.17,34.17,33.55,32.16,29.64,29.50,29.18,29.13,29.00,28.65,27.90,26.83,25.52,24.98,22.04,21.86,19.77,16.28,14.57.LC-MS(ESI)[M+H]+＝490.
Example 95
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (trifluoromethyl) -2, 3-dihydro-1H-indol-5-yl ] butanamide
The first step compound 5-nitro-1H-indole (1 g,6.167mmol,1.0 eq) was added to a reaction flask containing trifluoroacetic acid (2 mL), cooled to 0℃and stirred, triethylsilane (1.31 g,6.167mmol,1.0 eq) was slowly added dropwise, and reacted for 1 hour at room temperature; TLC (dichloromethane: methanol=20:1) monitored the reaction was complete. Dichloromethane (30 mL) was added to the reaction solution, washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=10:1) to give 5-nitro-2, 3-dihydro-1H-indole (95-1) (0.8 g,4.873mmol, 79%) as a white solid, LC-MS (ESI) [ m+h ] + =165.1
The second step of adding 5-nitro-2, 3-dihydro-1H-indole (95-1) (200 mg,1.218mmol,1.0 eq) into a reaction bottle containing N, N-dimethylformamide (5 mL), cooling to 0 ℃, stirring, slowly adding sodium hydrogen (116.96 mg,4.873mmol,4.0 eq), reacting at room temperature for 0.5 hours, adding 1.5mL (25%wt) of an N, N-dimethylformamide solution of trifluoroiodomethane, heating to 80 ℃, and reacting for 18 hours; TLC (dichloromethane: methanol=20:1) monitored the reaction. Dichloromethane (30 mL) was added to the reaction solution, washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (dichloromethane: methanol=10:1) to give the product 5-nitro-1- (trifluoromethyl) -2, 3-dihydro-1H-indole (95-2) (141.42 mg, 0.09 mmol, 50%) as a white solid LC-MS (ESI) [ m+h ] + =233
The third step, 5-nitro-1- (trifluoromethyl) -2, 3-dihydro-1H-indole (95-2) (100 mg,0.431mmol,1.0 eq) was added to a reaction flask containing methanol (5 mL), 10% palladium on carbon (20 mg) was added, hydrogen was replaced, and the reaction was carried out for 2 hours; TLC (dichloromethane: methanol=20:1) monitored reaction was complete; direct filtration and spin-drying of the organic solvent gave the product 1- (trifluoromethyl) -2, 3-dihydro-1H-indol-5-amine (95-3) (80 mg, 0.390 mmol, 91.86%) as a white solid LC-MS (ESI) [ M+H ] + =203.1
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (52.24 mg,0.150mmol,1 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (2 mL), and after adding (7-azobenzotriazole) -N, N, N\' -tetramethyl urea hexafluorophosphate (113.98 mg,0.300mmol,2 eq) and ethyl [ di (propan-2-yl) ] amine (58.12 mg,0.450mmol,3.0 eq), stirring was continued at room temperature (25 ℃) for 0.5hr, followed by adding 1- (trifluoromethyl) -2, 3-dihydro-1H-indol-5-amine (95-30 mg, 1.150 mmol, 1.1500 hr). TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (trifluoromethyl) -2, 3-dihydro-1H-indol-5-yl ] butanamide (95) (20 mg,0.038mmol, 25.14%) as a white solid .1H NMR(400MHz,CDCl3)δ7.58(s,1H),7.04(d,J＝8.7Hz,2H),6.94(d,J＝8.3Hz,1H),3.71(t,J＝8.3Hz,2H),3.22(dd,J＝11.5,4.6Hz,1H),3.09(t,J＝8.3Hz,2H),2.38–2.26(m,2H),2.15(dd,J＝25.6,9.7Hz,3H),2.04–1.90(m,3H),1.87–1.75(m,2H),1.72–1.64(m,4H),1.61(s,3H),1.43–1.31(m,2H),1.07(ddd,J＝18.7,13.5,7.9Hz,2H),0.97(d,J＝6.0Hz,3H),0.89–0.84(m,2H),0.80(d,J＝10.3Hz,7H).13C NMR(101MHz,CDCl3)δ171.14,133.00,132.39,131.92,127.80,119.40,117.87,112.43,112.40,79.11,54.30,53.96,48.17,38.83,37.84,37.45,36.96,36.53,33.58,32.12,28.48,28.36,28.06,27.65,24.36,21.93,21.86,19.66,15.37,14.30.LC-MS(ESI)[M+H]+＝533.3.
Example 96
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (9-hydroxynonyl) propionamide
In the first step, 2- (9-hydroxynonyl) isoindole-1, 3-dione (70-1) (0.5 g,1.73mmol,1.0 eq) was dissolved in ethanol (20 mL), hydrazine hydrate (0.42 mL,8.64mmol,5.0 eq) was added, the reaction was stirred at 80℃for 2h, and TLC (dichloromethane: methanol=10:1) showed completion of the reaction. Concentrating after the reaction, extracting with chloroform (20 mL), washing with water (20 mL. Times.2), drying with anhydrous Na 2SO4, and directly feeding crude 9-amino-non-1-ol (96-1) into the next step .1H NMR(400MHz,MeOD)δ3.44(t,J＝6.6Hz,2H),2.57–2.47(m,2H),1.51–1.33(m,4H),1.21(d,J＝17.6Hz,10H),1.05(d,J＝6.2Hz,1H).
In a second step 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (100.0 mg,0.22mmol,1.0 eq), HATU (101.6 mg,0.27mmol,1.2 eq) and N, N-diisopropylethylamine (37.4 mg,0.29mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (3.0 mL), and after stirring for 2min the reaction 9-amino-non-1-ol (96-2) (46.17 mg,0.29mmol,1.3 eq) was added and the reaction mixture was stirred overnight at room temperature. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.3). The organic layer was dried over anhydrous sodium sulfate and the crude was purified by passing methanol=15:1 through a column to give 3- [ (4 as,4 br) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (9-hydroxynonyl) propanamide (90 mg, purity 90%, yield 52.3%) which was directly taken to the next step.
In the fifth step, 3- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (9-hydroxynonyl) propionamide (96-2) (90 mg,0.15mmol,1.0 eq) was dissolved in tetrahydrofuran (2 mL), a solution of TBAF (5.0 mL,3.0mmol,32.8 eq) in tetrahydrofuran was added, and the reaction solution was stirred overnight at 60 ℃. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (30 mL. Times.2), and washed with water (20 mL. Times.4). The organic layer was dried over anhydrous sodium sulfate and the crude was purified with dichloromethane, methanol=10:1, and concentrated to give 3- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (9-hydroxynonyl) propanamide (96) as a white solid (33 mg, 96.2% purity, yield 43.6％).1H NMR(400MHz,CDCl3)δ5.46(s,1H),3.57(t,J＝6.6Hz,2H),3.20–3.09(m,3H),2.43–2.14(m,3H),2.13–2.02(m,2H),1.99–1.79(m,3H),1.70–1.55(m,10H),1.52–1.46(m,3H),1.44–1.36(m,2H),1.29–1.21(m,10H),1.01–0.87(m,5H),0.80(dd,J＝16.0,8.4Hz,2H),0.73(d,J＝9.3Hz,6H).13C NMR(101MHz,CDCl3)δ172.94,131.89,128.53,79.05,77.34,77.23,77.03,76.71,63.02,54.29,53.94,39.53,38.83,37.88,36.93,36.51,35.56,33.57,32.75,32.11,29.66,29.43,29.30,29.16,28.05,27.62,26.87,25.68,25.15,21.90,21.79,19.59,15.38,14.29.LC-MS(ESI)[M+H]+＝476.4.
Example 97
Preparation of the Compound N-decyl-3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-methylpropanamide
Reference example 63 by converting bromoethane to iodomethane, N-decyl-3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N-methylpropanamide is obtained (97).1H NMR(400MHz,CDCl3)δ3.38–3.11(m,3H),2.90(s,3H),2.36-2.13(m,4H),2.10-1.86(m,3H),1.75–1.66(m,6H),1.65–1.58(m,3H),1.56(d,J＝8.0Hz,3H),1.54–1.44(m,2H),1.32(dd,J＝13.0,3.5Hz,1H),1.29–1.19(m,13H),1.05-0.96(m,2H),0.94(d,J＝7.4Hz,1H),0.92(s,2H),0.89–0.79(m,5H),0.78-0.76(m,3H),0.74(s,2H).13C NMR(101MHz,CDCl3)δ172.85,132.35,127.93,79.07,77.35,77.03,76.71,54.33,53.98,50.02,47.82,38.83,38.36,36.96,36.52,35.37,33.47,32.75,32.15,31.89,29.59,29.55,29.31,28.06,27.64,26.92,25.25,24.81,22.67,21.94,21.78,19.51,15.38,14.29,14.11.LC-MS(ESI)[M+H]+＝488.45
Example 98
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (trifluoromethyl) indol-5-yl ] butanamide
The first step compound 5-nitro-1- (trifluoromethyl) -2, 3-dihydro-1H-indole (95-2) (100 mg,0.431mmol,1.0 eq) was added to a reaction flask containing carbon tetrachloride (5 mL), 1-bromotetrahydropyrrole-2, 5-dione (153.43 mg,0.862mmol,2.0 eq) was slowly added, the temperature was raised to 80℃and the reaction was carried out for 18 hours. TLC (dichloromethane: methanol=20:1) was monitored completely. Dichloromethane (30 mL) was added to the reaction solution, washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (dichloromethane: methanol=10:1) to give 3-bromo-5-nitro-1- (trifluoromethyl) indole (98-1) (80 mg,0.259mmol, 60.1%) as a white solid .1H NMR(400MHz,CDCl3)δ8.56(d,J＝2.2Hz,1H),8.31(dd,J＝9.1,2.2Hz,1H),7.69(dd,J＝9.1,1.5Hz,1H),7.55(s,1H).
The fourth step compound 3-bromo-5-nitro-1- (trifluoromethyl) indole (98-1) (80 mg, 0.299 mmol,1.0 eq) was added to a reaction flask containing methanol (5 mL), 10% palladium on carbon (16 mg) was added, hydrogen was replaced, and the reaction was carried out for 2 hours; TLC (dichloromethane: methanol=20:1) monitoring, with product formation; direct filtration and spin-drying of the organic solvent gave the product 1- (trifluoromethyl) indol-5-amine (98-2) (50mg,0.250mmol,96.5 0%) as a white solid LC-MS (ESI) [ m+h ] + =201.1
The fifth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (52.24 mg,0.150mmol,1 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (2 mL), and after adding (7-azobenzotriazole) -N, N, N\' -tetramethyl urea hexafluorophosphate (113.98 mg,0.300mmol,2 eq) and ethyl [ di (propan-2-yl) ] amine (58.12 mg,0.450mmol,3.0 eq), stirring was continued at room temperature (25 ℃) for 0.5hr, and then 1- (trifluoromethyl) indol-5-amine (30 mg,0.150 eq, 1.1 hr). TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (trifluoromethyl) indol-5-yl ] butyramide (98) (20 mg,0.038mmol, 25.14%) as a white solid .1H NMR(400MHz,CDCl3)δ7.98(s,1H),7.51(d,J＝9.5Hz,1H),7.28(d,J＝2.8Hz,1H),7.17(s,1H),6.62(d,J＝3.5Hz,1H),3.22(dd,J＝11.5,4.6Hz,1H),2.37(dq,J＝15.7,7.8Hz,2H),2.15(d,J＝7.7Hz,2H),1.99(d,J＝9.0Hz,2H),1.90(d,J＝13.6Hz,2H),1.78–1.66(m,4H),1.62(s,3H),1.59(s,2H),1.40–1.30(m,2H),1.05(ddd,J＝16.7,12.2,4.6Hz,2H),0.97(s,3H),0.94–0.84(m,3H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ171.25,133.02,132.41,130.78,130.22,127.80,123.84,117.49,112.88,112.01,107.08,79.11,54.30,53.96,38.83,37.84,37.55,36.97,36.53,33.58,32.13,29.32,28.49,28.05,27.65,24.38,21.93,21.86,19.67,15.37,14.30.LC-MS(ESI)[M+H]+＝531.3.
Example 99
Preparation of the Compound N-decyl-5- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanamide
In the first step, 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (300 mg,0.69mmol,1.0 eq) and ethoxyformylmethylene triphenylphosphine (724.5 mg,2.08mmol,3.0 eq) were dissolved in dichloromethane (15 mL) and the reaction was stirred at room temperature for 1.5hrs. TLC (petroleum ether: ethyl acetate=10:1) showed the reaction was complete. Concentrating the reaction liquid, and then carrying out column chromatography on petroleum ether: ethyl acetate=20:1 afforded (2E) -5- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pent-2-enoic acid ethyl ester (99-1) as a white solid (330 mg, purity 90%, yield 85.2％).1H NMR(400MHz,CDCl3)δ7.05–6.89(m,1H),5.82(d,J＝15.7Hz,1H),4.18(dt,J＝7.1,6.1Hz,2H),3.19(dd,J＝4.0Hz,1H),2.30-2.05(m,4H),2.04-1.85(m,3H),1.77–1.48(m,10H),1.40–1.24(m,5H),1.10-1.02(m,2H),0.92–0.86(m,13H),0.83–0.78(m,4H),0.76(d,J＝7.2Hz,3H),0.04(d,J＝8.2Hz,6H).
In a second step, (2E) -5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pent-2-enoic acid ethyl ester (99-1) (300 mg,0.60mmol,1.0 eq) was dissolved in THF (20 mL) and Pd/C10% (60 mg) was added under nitrogen. The reaction solution was replaced with hydrogen 3 times. The reaction mixture was stirred at room temperature for 1hrs under hydrogen atmosphere. 1 H NMR showed the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give ethyl 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] pentanoate (99-2) as a white solid (170 mg, purity 90%, yield 50.8%), the crude product was directly fed to the next step .1H NMR(400MHz,CDCl3)δ4.10(q,J＝7.1Hz,2H),3.15(dd,J＝4.1Hz,1H),2.27(t,J＝7.5Hz,2H),2.10-1.80(m,5H),1.72–1.58(m,6H),1.56(s,3H),1.35-30(m,3H),1.28–1.17(m,7H),1.08–0.93(m,3H),0.85-0.81(m,12H),0.77(s,3H),0.76–0.70(m,4H),0.01(t,J＝2.8Hz,6H).
In a third step, ethyl 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanoate (99-2) (230 mg,0.46mmol,1.0 eq) was dissolved in methanol (methanol) (4 mL) and aqueous NaOH (2 mL,10 mmol) was added at room temperature. The reaction solution was stirred at 45℃for 3hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was concentrated, extracted with ethyl acetate (20 mL), washed with water (15 mL. Times.3), and dried over anhydrous sodium sulfate. The organic layer was concentrated to give 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanoic acid (99-3) as a white solid (190 mg, purity 90%, yield 78.7%), the crude product was directly fed to the next step .1H NMR(400MHz,CDCl3)δ3.15(dd,J＝11.2,4.5Hz,1H),2.33(t,J＝7.4Hz,2H),2.10–1.80(m,5H),1.72–1.45(m,10H),1.44–1.21(m,4H),1.08–0.92(m,3H),0.89–0.83(m,13H),0.82–0.76(m,4H),0.73(d,J＝7.2Hz,3H),0.00(d,J＝2.4Hz,6H).
In the fourth step, 5- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanoic acid (99-3) (150 mg,0.32mmol,1.0 eq), HATU (143 mg,0.38mmol,1.2 eq) and N, N-diisopropylethylamine (52.8 mg,0.41mmol,1.3 eq) were dissolved in DMF (N, N-dimethylformamide) (4 mL), and after stirring for 2min, the reaction decylamine (41.8 mg,0.40mmol,1.3 eq) was added and the reaction mixture was stirred at room temperature for 1h. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1 column chromatography gives N-decyl-5- [ (4 br,7 s) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanamide (99-4) (130 mg, purity 90%, yield) 60.2％).1H NMR(400MHz,CDCl3)δ5.56(s,1H),3.18(d,J＝25.9Hz,3H),2.12(d,J＝44.6Hz,4H),1.92(d,J＝50.2Hz,3H),1.69–1.42(m,12H),1.30(s,2H),1.29–1.20(m,16H),0.92(d,J＝22.1Hz,3H),0.88–0.82(m,16H),0.77(s,3H),0.76–0.65(m,4H),0.00(d,J＝2.7Hz,6H).
In the fifth step, N-decyl-5- [ (4 aS,4 bR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanamide (99-4) (100 mg,0.16mmol,1.0 eq) was dissolved in THF (tetrahydrofuran) (1 mol/L,5 mL) of TBAF, and the reaction mixture was stirred at 80℃for 5hrs. TLC (petroleum ether: ethyl acetate=5:1) showed the reaction was complete. The reaction mixture was extracted with ethyl acetate (20 mL. Times.2), and washed with water (30 mL. Times.5). The organic layer was dried over anhydrous sodium sulfate and the crude product was petroleum ether: ethyl acetate=10:1, and concentrated to give N-decyl-5- [ (4 as,4 br) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] pentanamide (99) (26 mg, purity 88%, yield) as a colorless oil 26.2％).1H NMR(400MHz,CDCl3)δ5.37(s,1H),3.25–3.11(m,3H),2.17–1.77(m,7H),1.69–1.50(m,11H),1.47–1.32(m,4H),1.30-1.15(m,17H),1.05-0.90(m,5H),0.84–0.65(m,10H).13C NMR(101MHz,CDCl3)δ173.01,133.03,126.96,79.11,77.35,77.23,77.03,76.71,54.31,53.98,39.56,38.83,38.03,36.97,36.91,36.52,33.51,32.16,31.90,29.71,29.56,29.32,28.90,28.34,28.07,27.65,26.95,26.10,22.69,21.97,21.86,19.57,15.39,14.29,14.12.LC-MS(ESI)[M+H]+＝502.5.
Example 100
Preparation of Compound 100
In a first step, 8-phenyloct-1-ol (200 mg,0.97 mmol), pyridine chlorochromate (417.74 mg,1.94 mmol) and solvent dichloromethane (10 mL) were added to a 50mL three-necked flask under nitrogen, and the reaction solution was stirred at room temperature for 1hr. LC-MS monitored the reaction. After completion of the reaction, quenched with water, then extracted with ethyl acetate (15 ml x 3), the organic phase was washed twice with saturated brine (10 ml x 2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give the crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 20:1) to give 8-phenyloctanal (100-1) (100 mg,0.441mmol, yield: 45.45%) as pale yellow oil compound .1H NMR(400MHz,DMSO)δ9.65(t,J＝1.5Hz,1H),7.26(t,J＝7.4Hz,2H),7.16(dd,J＝12.2,7.1Hz,3H),2.65–2.52(m,2H),2.40(td,J＝7.2,1.5Hz,2H),1.63–1.45(m,4H),1.25(d,J＝18.6Hz,6H).
In a second step, 8-phenyloctanal (100-1) (50 mg,0.245 mmol), (1-diazonium-2-oxypropylene) dimethyl phosphonate (94.13 mg,0.490 mmol), potassium carbonate (67.72 mg,0.490 mmol) and solvent methanol (10 mL) were added to a 50mL three-necked flask under nitrogen, and the reaction stirred at room temperature for 8hr. LC-MS detection, after the reaction was completed, the reaction solution was diluted with ethyl acetate, and washed with water and saturated brine. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was taken up in silica gel (petroleum ether: ethyl acetate=20:1) and passed through a column, and the sample was collected to give the product non-8-alkynylbenzene (100-2) (25 mg,0.112mmol, yield: 45.90%) as a pale yellow oily compound .1H NMR(400MHz,DMSO)δ7.26(t,J＝7.4Hz,2H),7.16(d,J＝12.6,7.1Hz,3H),2.73(t,J＝2.6Hz,1H),2.62–2.53(m,2H),2.13(t,J＝6.9,2.6Hz,2H),1.61–1.49(m,2H),1.45–1.39(m,2H),1.31(d,J＝17.9,7.5Hz,6H).
In the third step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-azidopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (136-4) (30 mg,0.065 mmol), non-8-alkynylbenzene (100-2) (18.23 mg,0.098 mmol), sodium ascorbate (7.73 mg,0.039 mmol), copper sulfate pentahydrate (3.25 mg,0.013 mmo) were dissolved in t-butanol (2 mL) and H 2 O (2 mL) and stirred at room temperature overnight, and after completion of the reaction, the reaction was diluted with ethyl acetate and washed with saturated brine by LC-MS. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was passed through a column with (petroleum ether: ethyl acetate/5:1) to collect a sample to give the product 1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } -4- (6-phenylhexyl) -1,2, 3-triazacyclopenta-ne (100-3) (15 mg,0.022mmol, yield: 33.80%) as a pale yellow solid compound.
The fourth step was to dissolve the compound 1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -4- (7-phenylheptyl) -1,2, 3-triazacyclopenta-ne (100-3) (30 mg,0.045 mmol) in tetrahydrofuran (5 mL), then to add tetrabutylammonium fluoride (1M/tetrahydrofuran) (1.5 mL), displace nitrogen, warm to 50℃stirring overnight, LC-MS detect, quench with water after the reaction was completed, then extract with ethyl acetate (30 mL), wash the organic phase twice with saturated brine, dry with anhydrous sodium sulfate, filter, spin-dry to give the crude product, then separate and purify the crude product via preparative plate (petroleum ether); ethyl acetate=2:1) to give (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- {3- [4- (7-phenylheptyl) -1,2, 3-triazacyclopent-1-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (100) (15 mg,0.026mmol, yield: 57.44%) as a white solid compound .1H NMR(400MHz,DMSO)δ7.84(d,J＝4.8Hz,1H),7.27-7.24(m,2H),7.19–7.10(m,3H),4.26(t,J＝6.4Hz,2H),3.03–2.95(m,1H),2.578-2.53(m,4H),1.99–1.72(m,7H),1.67–1.40(m,13H),1.34–1.22(m,8H),0.99–0.91(m,2H),0.87(d,J＝5.7Hz,3H),0.79–0.70(m,6H),0.67(s,3H).13C NMR(101MHz,DMSO)δ147.26,142.74,132.25,128.69,128.66,127.39,126.04,122.14,77.35,54.31,53.90,49.68,38.90,37.99,37.01,36.50,35.60,33.46,32.05,31.46,29.52,29.23,29.09,29.05,28.88,28.62,27.88,26.01,25.44,22.03,21.83,19.63,16.25,14.53.LC-MS(ESI)[M+1]+＝546.4.
Example 101
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-undecylpropionamide
Reference example 27 (1-methylindol-5-yl) methanamine was converted to undecylamine to give the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N-undecylpropionamide (101).1H NMR(400MHz,CDCl3)δ5.45(s,1H),3.25-3.20(m,3H),2.50–2.07(m,5H),2.05–1.85(m,3H),1.79–1.62(m,5H),1.60-1.53(m,3H),1.53–1.41(m,3H),1.37(dd,J＝13.0,3.5Hz,1H),1.28-1.25(m,16H),1.08–0.99(m,2H),0.97(s,3H),0.92–0.84(m,5H),0.81(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ172.83,131.92,128.49,79.06,77.33,77.02,76.70,54.30,53.95,39.55,38.83,37.89,36.94,36.51,35.60,33.57,32.12,31.91,29.69,29.59,29.54,29.32,28.04,27.63,26.95,25.14,22.68,21.91,21.80,19.57,15.36,14.29,14.11.
LC-MS(ESI)[M+H]+＝488.
Example 102
Preparation of the Compound N- (4-fluoro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
In the first step, 5-bromo-4-fluoro-1H-indole (100 mg,0.467 mmol) was dissolved in N, N-dimethylformamide (3 mL), then the mixture was cooled to 0℃under nitrogen, sodium hydride (22.43 mg,0.934 mmol) was added to the solution, stirred at 0℃for half an hour, methyl iodide (0.058 mL,0.934 mmol) was added dropwise, and the mixture was warmed to room temperature and stirred for 16 hours. After the reaction was completed, 30mL of water was added, extraction was performed with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product, and the crude product was purified by a preparation plate (developing solvent: petroleum ether: ethyl acetate=5/1) to obtain a pale yellow solid, 5-bromo-4-fluoro-1-methylindole (102-1)(95mg,0.396mmol,84.70％).1H NMR(400MHz,DMSO)δ7.44(d,J＝3.1Hz,1H),7.32(m,2H),6.52(d,J＝3.1Hz,1H),3.81(s,3H).LC-MS(ESI)[M+H]+＝228.0.
In the second step, 5-bromo-4-fluoro-1-methylindole (102-1) (60 mg,0.263 mmol), benzophenone imine (71.52 mg, 0.015 mmol), potassium tert-butoxide (59.02 mg,0.526 mmol), 1 '-binaphthyl-2, 2' -bis-diphenylphosphine (BINAP) (245.96 mg, 0.399 mmol) were dissolved in toluene (3 mL), and tris (dibenzylideneacetone) dipalladium (24.08 mg,0.026 mmol) was added thereto under nitrogen atmosphere, followed by stirring at 100℃for 16 hours. After completion of the reaction, the crude product was dried by spin-drying in vacuo and purified by column chromatography (petroleum ether: ethyl acetate=3/1) to give 5- [ (diphenylmethylene) amino ] -4-fluoro-1-methylindole (102-2) (100 mg,0.122mmol, 46.30%).
LC-MS(ESI)[M+H]+＝329.1.
In the third step, 5- [ (diphenylmethylene) amino ] -4-fluoro-1-methylindole (102-2) (20 mg,0.061 mmol) was dissolved in methanol (15 mL), and a solution of 1, 4-dioxane (0.5 mL) of hydrochloric acid (2.22 mg,0.061 mmol) was added under ice bath, stirred at room temperature for two hours, and monitored by LC-MS. After the reaction is completed, the reaction solution is dried by spinning, 1mL of methanol solution of ammonia is added under ice bath, after being fully stirred, crude products are obtained by spinning, the crude products are sampled by a silica gel plate and are unfolded by a preparation plate (developer: petroleum ether: ethyl acetate=1/2) to obtain the product 4-fluoro-1-methylindole-5-amine (102-3)(10mg,0.058mmol,95.00％).1H NMR(400MHz,DMSO)δ7.17(d,J＝3.0Hz,1H),6.98(d,J＝8.5Hz,1H),6.70(t,J＝8.5Hz,1H),6.23(d,J＝3.0Hz,1H),4.45(s,2H),3.69(s,3H).LC-MS(ESI)[M+H]+＝165.1.
In a fourth step 4-fluoro-1-methylindole-5-amine (102-3) (12 mg,0.073 mmol), 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (22 mg,0.063 mmol) and 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (36.00 mg,0.095 mmol) were dissolved in N, N, N-diisopropylethylamine (0.031 mL,0.189 mmol) was added after N-dimethylformamide (2 mL), the reaction was stirred at room temperature for 16 hours, and the crude product was purified via a preparative plate (dichloromethane/methanol=20/1) to give the product N- (4-fluoro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide (102)(22mg,0.044mmol,69.75％).1H NMR(400MHz,DMSO)δ9.45(s,1H),7.35(d,J＝3.1Hz,1H),7.22(dd,J＝10.4,7.6Hz,2H),6.46(d,J＝3.1Hz,1H),4.31(d,J＝5.1Hz,1H),3.79(s,3H),3.00(d,J＝6.7Hz,2H),2.31(t,J＝7.1Hz,3H),2.17(d,J＝9.8Hz,2H),2.06(d,J＝8.7Hz,2H),1.94(s,2H),1.66(d,J＝13.3Hz,3H),1.57(d,J＝13.8Hz,5H),1.50(s,3H),1.25(d,J＝9.6Hz,3H),0.88(s,3H),0.77(s,3H).13C NMR(101MHz,DMSO)δ171.78,141.37,136.95,135.10,133.13,132.50,130.92,129.11,126.96,123.60,120.57,114.05,105.93,99.99,96.51,81.83,77.39,62.68,54.38,54.11,54.01,38.09,37.35,37.06,36.85,36.54,36.15,36.00,33.96,33.53,33.24,33.09,32.23,31.97,28.64,28.54,27.90,27.33,24.80,22.70,22.09,22.02,21.93,20.14,19.81,16.89,16.27,15.68,14.58.19F NMR(376MHz,DMSO)δ-131.96.LC-MS(ESI)[M+H]+＝495.3.
Example 103
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazapentan-5-yl) amino ] methanamide
The first step compound 3- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylhydrophenanthrene ] propanoic acid (I) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing toluene (2 mL), diphenyl azide phosphate (64.39 mg,0.234mmol,1.05 eq) and triethylamine (0.065 mL, 0.268 mmol,2.0 eq) were added and stirred at room temperature (90 ℃) for 1hr, the reaction mixture was cooled to room temperature, tetrahydrofuran (2 mL) was added, and 1-methylbenzo [ d ] [1,2,3] triazacyclopenta-5-amine (33.04 mg,0.223mmol,1.0 eq) was further stirred for 18hr. LC-MS monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by preparative TLC (petroleum ether: ethyl acetate=1:1) to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] methanamide (103-1) (30 mg,0.045mmol, 20.39%) as a pale yellow solid. LC-MS (ESI) [ m+h ] + = 462.30
In the second step N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] methanamide (103-1) (30 mg,0.051mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.505 mL,0.505mmol,10.0 eq) was added and then stirred at 50℃for 18hr; TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by preparative TLC (dichloromethane: methanol=10:1), separating to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-N-5-yl) amino ] methanamide (103) (6 mg,0.013mmol, 24.76%) as a white solid .1H NMR(400MHz,CDCl3)δ7.97(s,1H),7.55(dd,J＝8.9,3.3Hz,1H),7.39(m,1H),4.20(d,J＝3.7Hz,3H),3.05(m,1H),2.25(dd,J＝35.1,10.0Hz,3H),1.90(d,J＝28.6Hz,3H),1.65(d,J＝15.9Hz,3H),1.58(s,3H),1.55(s,2H),1.36(dd,J＝25.5,13.1Hz,2H),1.20(s,3H),0.99(m,2H),0.86(d,J＝3.5Hz,3H),0.78(s,1H),0.76(d,J＝3.4Hz,3H),0.70(dd,J＝7.9,3.6Hz,3H).13C NMR(101MHz,CDCl3)δ137.00,130.08,129.08,121.76,109.82,106.08,78.37,54.44,54.22,38.53,37.94,36.90,36.23,33.29,32.18,29.16,27.24,26.93,21.70,18.51,14.72,13.39.LC-MS(ESI)[M+H]+＝480.30.
Example 104
Preparation of the Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [4- (1-methylindol-5-yl) -1,2, 3-triazacyclopentan-1-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In the first step, the compound 5-iodo-1H-indole (1.00 g,4.114mmol,1.0 eq) was dissolved in tetrahydrofuran (10 mL), then replaced with nitrogen, cooled to-78 ℃, then sodium hydrogen (0.20 g,4.937mmol,1.2 eq) was added thereto, stirred for 1H, then methyl iodide (0.284 mL,6.172mmol,1.5 eq) was added thereto, stirred at room temperature overnight, and TLC (petroleum ether: ethyl acetate=10:1) monitored for reaction. After the completion of the reaction, water was added thereto for quenching, followed by extraction with ethyl acetate (50 mL), and the organic phase was washed twice with saturated brine (50 mL x 2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give 5-iodo-1-methylindole (104-1) (1 g,3.89mmol, yield: 94.55%) as a white solid compound .1H NMR(400MHz,CDCl3)δ7.95(d,J＝1.5Hz,1H),7.46(dd,J＝8.6,1.6Hz,1H),7.10(d,J＝8.6Hz,1H),7.01(d,J＝3.1Hz,1H),6.47–6.34(m,1H),3.77(s,3H).
In a second step, the compound 5-iodo-1-methylindole (104-1) (75 mg,0.292mmol,1.0 eq), trimethylethynyl silicon (0.062 mL,0.438mmol,1.5 eq), ditriphenylphosphine palladium dichloride (13.62 mg,0.018mmol,0.06 eq), cuprous iodide (14.81 mg,0.047mmol,0.16 eq) was dissolved in dichloromethane (1 mL) and triethylamine (0.5 mL), then replaced with nitrogen, and stirred at room temperature for 3h. After completion of LC-MS monitoring, the mixture was dried by spin-drying and then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give 1-methyl-5- [ (trimethylsilyl) ethynyl ] indole (104-2) (48 mg,0.21mmol, yield: 72.35%) as an oily compound .1H NMR(400MHz,CDCl3)δ7.79(s,1H),7.33(d,J＝8.4Hz,1H),7.23(d,J＝8.5Hz,1H),7.06(d,J＝3.1Hz,1H),6.45(d,J＝3.1Hz,1H),3.78(s,3H),0.26(s,9H).
In the third step, compound 1-methyl-5- [ (trimethylsilyl) ethynyl ] indole (104-2) (48 mg,0.211mmol,1.0 eq) was dissolved in tetrahydrofuran (1 mL) and methanol (2 mL), then sodium hydroxide (33.78 mg,0.844mmol,4.0 eq) was added thereto, stirred at room temperature for 2 hours, and the reaction was monitored by tlc (petroleum ether: ethyl acetate=10:1). Water was added to quench, followed by extraction with ethyl acetate (50 mL), and the organic phase was washed twice with saturated brine (50 mL. Times.2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give 5-ethynyl-1-methylindole (104-3) (32 mg,0.206mmol, yield: 97.68%) as crude product without further purification. LC-MS (ESI) [ m+h ] + =156.2.
In a fourth step, the compound 4-methylbenzenesulfonic acid-3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (II-1) (150 mg,0.255mmol,1.0 eq) was dissolved in dimethyl sulfoxide (3 mL), then sodium azide (49.67 mg,0.76 mmol,3.0 eq) was added thereto, nitrogen was displaced, and then heated to 50℃and stirred overnight, and TLC (petroleum ether: ethyl acetate=10:1) monitored the reaction. After completion of the reaction, water was added thereto for quenching, followed by extraction with ethyl acetate (50 mL), and the organic phase was washed twice with saturated brine (50 mL. Times.2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-azidopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (104-4) (115 mg,0.25mmol, yield: 98.21%) as an oily compound, the crude product was directly used for the next step .1H NMR(400MHz,CDCl3)δ3.20–3.15(m,1H),2.18–2.06(m,3H),1.97(dd,J＝21.1,8.9Hz,4H),1.70(m,4H),1.61(s,3H),1.33(m,6H),1.07–0.94(m,4H),0.88(d,J＝4.2Hz,12H),0.81(d,J＝3.7Hz,3H),0.75(s,3H),0.03(d,J＝2.8Hz,6H).
In a fifth step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-azidopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (104-4) (30 mg,0.065mmol,1.0 eq), 5-ethynyl-1-methylindole (104-3) (18.23 mg,0.117mmol,1.8 eq), sodium vitamin C (7.37 mg,0.037mmol,0.57 eq), copper sulfate pentahydrate (3.26 mg,0.013mmol,0.2 eq) were dissolved in t-butanol (1 mL) and H 2 O (1 mL) and stirred overnight at room temperature. LC-MS monitored the reaction. After completion of the reaction, water was added to dilute, ethyl acetate was added to extract (50 mL), the organic phase was washed twice with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified (petroleum ether: ethyl acetate=2:1) via a preparative plate to give 5- (1- {3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1,2, 3-triazapentan-4-yl) -1-methylindole (104-5) (15 mg,0.024, yield: 37.38%) as a solid compound. LC-MS (ESI) [ M-OTBS ] + =483.3.
In a sixth step, the compound 5- (1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1,2, 3-triazapentan-4-yl) -1-methylindole (104-5) (30 mg,0.049mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), then tetrabutylammonium fluoride (1M in tetrahydrofuran) (1.5 mL) was added thereto, nitrogen was replaced, and the mixture was warmed to 50℃overnight. LC-MS monitored the reaction. After completion of the reaction, water was added thereto for quenching, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then purified by preparative plate separation (petroleum ether: ethyl acetate=2:1) to give (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- {3- [4- (1-methylindol-5-yl) -1,2, 3-triazacyclopent-1-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrein-2-ol (104) (18 mg,0.033mmol, yield: 66.86%) as a solid compound, and freeze-dried to give a powdery compound .1H NMR(400MHz,CDCl3)δ8.08(s,1H),7.74(t,J＝3.5Hz,2H),7.37(d,J＝8.5Hz,1H),7.07(d,J＝3.1Hz,1H),6.52(d,J＝2.9Hz,1H),4.37(t,J＝7.1Hz,2H),3.82(s,3H),3.21(dd,J＝11.5,4.6Hz,1H),2.20–2.11(m,2H),2.05(dd,J＝12.5,3.2Hz,2H),2.00–1.87(m,4H),1.76(dd,J＝13.1,3.5Hz,2H),1.69–1.61(m,4H),1.57(s,3H),1.34(d,J＝6.2Hz,1H),1.09–1.01(m,2H),0.96(s,3H),0.91–0.85(m,2H),0.81(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ149.09,136.67,131.41,129.57,128.72,128.42,122.19,119.93,118.69,118.20,109.56,101.36,79.08,54.26,53.91,50.34,38.82,37.85,36.92,36.50,33.53,32.95,32.11,29.70,29.16,28.04,27.62,26.03,21.89,21.77,19.59,15.36,14.28.LC-MS(ESI)[M+1]+＝501.3.
Example 105
Preparation of the Compound N- [ 3-cyano-1- (pyrazin-2-ylmethyl) indol-5-yl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] butanamide
In a 50mL three-necked flask, under nitrogen protection, the reactant 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (59-3) (25 mg,0.072 mmol), 5-amino-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile (21.46 mg,0.086 mmol), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (27.27 mg,0.072 mmol), N, N-diisopropylethylamine (0.012 mL,0.072 mmol) and solvent DMF (3 mL) were added, and the reaction mixture was stirred at N2 for 8hr at room temperature. LC-MS monitored the reaction. The reaction mixture was added with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=1:0 to 10:1) to give N- [ 3-cyano-1- (pyrazin-2-ylmethyl) indol-5-yl ] -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide (105) (15 mg,0.023mmol, 32.46%) as a white solid .1H NMR(400MHz,DMSO)δ9.90(s,1H),8.70(s,1H),8.58(d,J＝4.1Hz,2H),8.37(s,1H),8.09(s,1H),7.57(d,J＝8.9Hz,1H),7.36(d,J＝9.0Hz,1H),5.67(s,2H),4.30(d,J＝4.9Hz,1H),2.99(d,J＝5.8Hz,1H),2.29(t,J＝6.5Hz,2H),2.16–1.81(m,6H),1.75–1.57(m,6H),1.49(s,3H),1.32(d,J＝14.2Hz,2H),0.97(dd,J＝20.3,9.8Hz,2H),0.81(d,J＝43.7Hz,9H),0.67(s,3H).13C NMR(101MHz,DMSO)δ171.56,151.78,144.93,144.78,144.01,138.26,135.01,132.99,132.11,127.89,127.03,117.20,116.36,112.30,108.86,84.51,77.39,54.37,53.98,49.63,38.92,37.98,37.05,36.72,36.53,33.53,32.22,28.62,27.90,24.52,22.07,21.88,19.83,16.25,14.56,0.57.LC-MS(ESI)[M+H]+＝580.3.
Example 106
Preparation of the Compound N- (3-cyano-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
Reference example 105, 5-amino-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile was changed to 5-amino-1-methylindole-3-carbonitrile (91-2) to give the compound N- (3-cyano-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (106).1H NMR(400MHz,DMSO)δ9.91(d,J＝8.2Hz,1H),8.20–8.12(m,1H),8.08(d,J＝7.7Hz,1H),7.54(t,J＝8.8Hz,1H),7.41(d,J＝8.7Hz,1H),4.31(d,J＝5.1Hz,1H),3.85–3.80(m,3H),2.98(s,1H),2.29(d,J＝6.9Hz,2H),2.09(d,J＝36.9Hz,4H),1.88(d,J＝27.6Hz,4H),1.61(s,3H),1.56(d,J＝7.8Hz,3H),1.30(s,2H),1.23(s,2H),0.93(s,2H),0.86(d,J＝8.5Hz,3H),0.79(s,1H),0.75(d,J＝8.3Hz,3H),0.67(d,J＝8.8Hz,3H).13C NMR(101MHz,DMSO)δ171.53,138.13,134.86,132.98,132.46,127.77,126.79,116.87,112.00,108.62,83.30,77.38,54.36,53.97,37.97,37.05,36.53,33.86,33.53,32.23,28.62,27.90,24.52,21.88,19.84,16.26,14.57.LC-MS(ESI)[M+1]+＝502.3.
Example 107
Preparation of the Compound 1- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -11-phenylundecan-3-one
In the first step, 1- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -11-phenylundecan-3-ol (10 mg,0.016mmol,1.0 eq) was dissolved in dichloromethane (5 mL), then dessert-martin oxidizer (13.61 mg,0.032mmol,2.0 eq) was added at room temperature, stirred at room temperature for 2h, tlc (petroleum ether: ethyl acetate=5:1) the reaction was monitored, when the reaction was completed saturated sodium bicarbonate was added thereto, then extracted with dichloromethane (50 mL), the organic phase was dried over anhydrous sodium sulfate, then filtered, and spin-dried to give a crude product which was purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give 1- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -11-phenylundec-3-one (8 mg, yield: 80.26%) as a solid compound .1H NMR(400MHz,CDCl3)δ7.23–7.19(m,2H),7.13–7.04(m,3H),3.11(dd,J＝11.1,4.6Hz,1H),2.62–2.44(m,2H),2.40–2.23(m,4H),2.18–1.94(m,2H),1.85(d,J＝13.8Hz,2H),1.60(dd,J＝22.9,15.5Hz,5H),1.52(s,3H),1.34–1.09(m,20H),0.82(s,9H),0.80(s,3H),0.74(s,3H),0.69(d,J＝6.4Hz,3H),0.02–-0.01(m,6H).
In a second step, the compound 1- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -11-phenylundecan-3-one (10 mg,0.016 mmol) was dissolved in tetrahydrofuran (3 mL), then tetrabutylammonium fluoride (2 mL) was added at room temperature, nitrogen was replaced, and the temperature was raised to 50℃and stirred overnight. LC-MS monitored the reaction. When the reaction was completed, water was added to effect the deactivation, followed by extraction with ethyl acetate, and the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give 1- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -11-phenylundecan-3-one (107) (6.2 mg, yield: 38.38%) as an oily compound .1H NMR(400MHz,CDCl3)δ7.28(d,J＝7.8Hz,2H),7.23–7.11(m,3H),3.22(dd,J＝11.4,4.6Hz,1H),2.66–2.57(m,2H),2.38(ddd,J＝20.5,15.6,5.0Hz,4H),1.96–1.62(m,10H),1.59(s,3H),1.56(s,3H),1.41–1.28(m,14H),1.00–0.95(m,3H),0.88(s,1H),0.81(s,3H),0.78(d,J＝9.0Hz,3H).13C NMR(101MHz,CDCl3)δ211.49,142.88,131.92,128.39,128.23,127.99,125.57,79.09,54.31,53.97,42.88,41.86,38.83,38.13,36.94,36.51,35.97,33.49,31.48,29.71,29.34,29.27,28.06,27.64,23.90,23.51,21.92,21.77,21.36,19.48,15.37,14.28.LC-MS(ESI)[M+1]+＝507.9.
Example 108
Preparation of Compound 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -1-oxobutylene } amino) -1-methylindole-3-carboxylic acid methyl ester
In the first step, the compound 1-methyl-5-nitroindole-3-carbonitrile (91-1) (100 mg,0.497mmol,1.0 eq) was dissolved in methanol (10 mL), then concentrated sulfuric acid (98%) (2 mL) was slowly added, then warmed to 60℃and stirred for 18 hours, and the reaction was detected by LC-MS. When the reaction was completed, the reaction solution was added to ice water, then slowly neutralized to weakly alkaline with sodium hydrogencarbonate, then extracted with methylene chloride, the organic phase was washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, then filtered and spin-dried to give a crude product, which was further purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 1:1) to give methyl 1-methyl-5-nitroindole-3-carboxylate (108-1) (55 mg,0.235mmol, yield: 47.25%) as a yellow solid compound .1H NMR(400MHz,MeOD)δ9.00(d,J＝2.0Hz,1H),8.28–8.15(m,2H),7.68(d,J＝9.1Hz,1H),3.96(d,J＝10.0Hz,6H).LC-MS(ESI)[M+H]+＝235.1
In a second step, the compound methyl 1-methyl-5-nitroindole-3-carboxylate (55 mg,0.235mmol,1.0 eq) was dissolved in methanol (10 mL), then palladium on carbon (24.99 mg,0.235 mmol) was added, hydrogen was replaced, and stirred at room temperature for 2h, and the reaction was detected by LC-MS. After the completion of the reaction, palladium on carbon was removed by filtration, followed by spin-drying to give methyl 5-amino-1-methylindole-3-carboxylate (108-2) (21 mg, yield: 43.79%), the crude product was used directly in the next reaction. LC-MS (ESI) [ m+h ] + =205.2
In a third step, compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (25 mg,0.054mmol,1.0 eq), methyl 5-amino-1-methylindole-3-carboxylate (108-2) (11.03 mg,0.054mmol,1.0 eq), HATU (30.81 mg,0.081mmol,1.5 eq) was dissolved in N, N-dimethylformamide (5 mL), then N, N-diisopropylethylamine (0.027 mL,0.162mmol,3.0 eq) was added thereto and stirred overnight at room temperature. LC-MS monitoring. After completion of the reaction, water quenching was added, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then purified by preparative chromatography (0.1% FA/MeCN/H 2 O) and then lyophilized to give methyl 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -1-oxybuty } amino) -1-methylindole-3-carboxylate (108) (20 mg, yield: 55.33%) as a white solid compound .1H NMR(400MHz,MeOD)δ8.21(d,J＝1.8Hz,1H),7.92(s,1H),7.53(dd,J＝8.8,2.0Hz,1H),7.41(d,J＝8.8Hz,1H),3.88(s,3H),3.86(s,3H),3.15(dd,J＝10.6,5.5Hz,1H),2.39(td,J＝7.2,2.5Hz,2H),2.26–2.11(m,3H),2.08–1.73(m,6H),1.69(dd,J＝12.9,6.4Hz,3H),1.64(s,3H),1.63–1.58(m,2H),1.42(dd,J＝12.9,3.3Hz,1H),1.13–1.01(m,2H),0.95(s,3H),0.85(s,3H),0.82(d,J＝10.9Hz,2H),0.78(s,3H).13C NMR(101MHz,MeOD)δ173.20,165.89,136.24,134.66,133.04,132.45,127.18,126.59,116.91,112.85,109.82,105.83,78.38,54.45,54.22,49.98,38.51,37.81,36.91,36.39,36.22,33.25,32.23,32.08,28.01,27.24,26.93,24.40,21.70,21.61,18.42,14.71,13.40.LC-MS(ESI)[M+H]+＝535.3.
Example 109
Preparation of the Compound N- (1, 6-dimethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
In the first step, the reaction iodine (1.67 g, 6.578mmol), silver sulfate (2.04 g, 6.578mmol) was added to the solvent ethanol (15 mL), the system was filled with nitrogen, placed in an ice bath, 3-methyl-4-nitroaniline (1 g, 6.578mmol) was dissolved in ethanol (15 mL) and added to the system, and the reaction mixture was stirred at room temperature under nitrogen for 5 hours, and monitored by LC-MS. After completion of the reaction, the reaction mixture was concentrated in vacuo, and water (40 mL) and ethyl acetate (3X 30 mL) were added thereto for extraction. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (eluent: petroleum ether: ethyl acetate=3/1) to give 2-iodo-5-methyl-4-nitroaniline (109-1) as a yellow solid (1.152 g,4.143mmol, 63.04%). LC-MS (ESI) [ m+h ] + =279.0
In the second step, 2-iodo-5-methyl-4-nitroaniline (109-1) (1.152 g,4.143 mmol), trimethylsilylacetylene (0.41 g,4.143 mmol), cuprous iodide (I) (0.08 g,0.414 mmol), bis (triphenylphosphine) palladium (II) dichloride (0.32 g,0.414 mmol), triethylamine (2.879 mL,20.716 mmol) were added to tetrahydrofuran (15 mL), and the reaction mixture stirred at room temperature under nitrogen for 3 hours and monitored by LC-MS. After completion of the reaction, water (60 mL) and ethyl acetate (3X 50 mL) were added thereto for extraction. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (developer: petroleum ether: ethyl acetate=10/1) to give 5-methyl-4-nitro-2- [ (trimethylsilyl) ethynyl ] aniline (109-2) as a yellow solid (900 mg,3.624mmol, 87.46%). LC-MS (ESI) [ m+h ] + =249.1
In the third step, the reaction mixture was stirred under nitrogen at 100℃for 5 hours and monitored by LC-MS, 5-methyl-4-nitro-2- [ (trimethylsilyl) ethynyl ] aniline (109-2) (940 mg,3.785 mmol) and copper (I) iodide (720.82 mg,3.785 mmol) were added to solvent N, N-dimethylformamide (10 mL). After completion of the reaction, water (60 mL) and ethyl acetate (3X 50 mL) were added thereto for extraction. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (developer: petroleum ether: ethyl acetate=30/1) to give 6-methyl-5-nitro-1H-indole (109-3) (1000 mg,2.838mmol, 74.98%) as a clear oil. LC-MS (ESI) [ m+h ] + =177.1
In the fourth step, 6-methyl-5-nitro-1H-indole (109-3) (100 mg, 0.618 mmol) was added to solvent N, N-dimethylformamide (3 mL), the reaction mixture was placed in an ice bath, the system was filled with nitrogen, sodium hydrogen (27.24 mg,1.135 mmol) was added at 0℃and stirred for 30 minutes, and then methyl iodide (161.13 mg,1.135 mmol) was added at 0℃and the reaction mixture was stirred at room temperature for 2 hours and monitored by LC-MS. After the reaction was completed, 30mL of water was added for quenching, and extracted with ethyl acetate (3X 25 mL) and concentrated in vacuo to give the crude product. Purifying the crude product by silica gel column (petroleum ether: ethyl acetate=3/1), chiral preparing to obtain yellow solid 1, 6-dimethyl-5-nitroindole (109-4)(35mg,0.184mmol,32.42％).1H NMR(400MHz,DMSO)δ8.37(s,1H),7.51(s,2H),6.63(d,J＝3.1Hz,1H),3.83(s,3H),2.65(s,3H).
In the fifth step, the reaction mixture was stirred at 80℃for 2 hours with 1, 6-dimethyl-5-nitroindole (109-4) (30 mg,0.158 mmol), iron powder (44.04 mg,0.789 mmol), ammonium chloride (84.37 mg,1.577 mmol) and solvent ethanol (3 mL), water (1 mL). LC-MS monitoring. After the reaction was completed, the mixture was filtered through celite, and rinsed 3 times with 3mL of ethanol. Concentration in vacuo afforded a brown solid. To the solid was added 30mL of water, extracted with ethyl acetate (3×25 mL), dried over anhydrous sodium sulfate and concentrated in vacuo to give crude product, which was purified by prep. plate (dichloromethane/methanol=15/1) to give 1, 6-dimethylindol-5-amine (109-5) (16 mg,0.100mmol, 63.31%). LC-MS (ESI) [ m+h ] + =161.1
In a sixth step, the reactant 1, 6-dimethylindol-5-amine (16 mg,0.100 mmol), 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (109-5) (34.81 mg,0.100 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (56.96 mg,0.150 mmol), N, N-diisopropylethylamine (0.050 mL,0.300 mmol) were added to solvent N, N-dimethylformamide (1 mL), and the reaction mixture was stirred overnight under nitrogen at room temperature and monitored by LC-MS. After completion of the reaction, water (15 mL) was added thereto, and the mixture was extracted with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=15/1) and prepared from formic acid to give N- (1, 6-dimethylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide as a white solid (109)(19.20mg,0.039mmol,38.90％).1H NMR(400MHz,)δ9.17(s,1H),7.37(s,1H),7.25–7.21(m,2H),6.31(d,J＝2.8Hz,1H),4.33(d,J＝5.0Hz,1H),3.73(s,3H),3.00(dd,J＝12.9,7.5Hz,1H),2.27(s,5H),2.01(ddd,J＝45.2,36.8,28.0Hz,6H),1.58(s,11H),1.39–1.21(m,3H),1.05–0.93(m,2H),0.87(s,3H),0.77(s,3H),0.68(s,3H).13C NMR(101MHz,CDCl3)δ171.70,132.55,129.00,127.00,117.04,110.21,100.95,79.13,54.30,53.98,38.84,37.91,37.36,36.97,36.54,33.58,32.89,32.14,28.73,28.07,27.65,24.78,21.94,21.87,19.71,18.86,15.39,14.31,0.02.LC-MS(ESI)[M+H]+＝491.3.
Example 110
Preparation of Compound (2S, 4aR,4bS,8aR,10 aR) -8- (3-hydroxy-11-phenylundecyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol
In the first step, magnesium chips (270.83 mg,5.571mmol,1.5 eq) and iodine (47.13 mg,0.186mmol,0.05 eq) were placed in a dry three-necked flask, then nitrogen was replaced, a small amount of a tetrahydrofuran solution of (8-bromooctyl) benzene (200 mg,0.742mmol,0.2 eq) was added, then the reaction was initiated by heating with a high temperature gun, then a tetrahydrofuran (10 mL) solution of (8-bromooctyl) benzene (800 mg,2.971mmol,0.8 eq) was slowly added dropwise, and after the dropwise addition, the mixture was heated and refluxed for one hour, and cooled to room temperature to obtain a solution of bromine (8-phenyloctyl) magnesium (110-1).
In a second step, compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (60 mg,0.139mmol,1.0 eq) was dissolved in tetrahydrofuran (5 mL), then magnesium (110-1) (81.39 mg,0.277mmol,2.0 eq) was added with bromine (8-phenyloctyl) at room temperature, followed by stirring for 30min, TLC (petroleum ether: ethyl acetate=5:1) and, after completion of the reaction, water was added thereto to quench, then extracted with ethyl acetate (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give 1- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -11-phenylundecan-3-ol (110-2) (30 mg, yield: 34.73%) as a solid compound .1H NMR(400MHz,CDCl3)δ7.25(d,J＝8.0Hz,2H),7.14(d,J＝6.6Hz,3H),3.52(s,1H),3.15(dd,J＝11.2,4.5Hz,1H),2.64–2.48(m,2H),2.20–2.05(m,2H),1.90(dd,J＝35.1,11.5Hz,3H),1.67(dd,J＝15.7,6.6Hz,3H),1.59(s,3H),1.39(dd,J＝22.2,13.5Hz,10H),1.27(s,12H),1.09–0.89(m,3H),0.85(d,J＝5.7Hz,12H),0.77(d,J＝2.2Hz,3H),0.73(d,J＝7.3Hz,3H),0.00(d,J＝2.8Hz,6H),0.00(d,J＝2.8Hz,6H).
In a third step, the compound 1- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -11-phenylundecan-3-ol (110-2) (32 mg,0.051mmol,1.0 eq) was dissolved in tetrahydrofuran (3 mL), then tetrabutylammonium fluoride (2 mL) was added at room temperature, nitrogen was displaced, heated to 50℃for stirring overnight, LC-MS detection, water was added thereto after completion of the reaction, then ethyl acetate (50 mL) was used for extraction, the organic phase was washed twice with saturated sodium chloride, then dried over anhydrous sodium sulfate, filtered, and dried by spin-drying to give a crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give an oily compound, and lyophilization to give (2 s,4ar,4bs,8ar,10 ar) -8- (3-hydroxy-11-phenylundecyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (110) (16.16 mg, yield: 56.9%) as a white solid .1H NMR(400MHz,MeOD)δ7.30–7.19(m,5H),7.13(dd,J＝12.9,7.1Hz,3H),3.48(d,J＝4.5Hz,1H),3.16(dd,J＝10.4,5.8Hz,1H),2.68–2.51(m,2H),2.38–2.09(m,3H),2.06–1.83(m,4H),1.74(ddd,J＝25.5,12.8,2.9Hz,5H),1.62(d,J＝5.4Hz,3H),1.55(s,2H),1.52–1.38(m,6H),1.32(dd,J＝20.7,7.0Hz,11H),1.11–1.01(m,2H),0.96(d,J＝5.6Hz,3H),0.84(d,J＝2.9Hz,3H),0.79(d,J＝7.3Hz,3H).13C NMR(101MHz,MeOD)δ142.56,133.11,132.82,127.98,127.85,126.53,126.31,125.22,78.37,71.42,71.03,54.47,54.28,54.24,38.53,38.22,37.89,36.99,36.93,36.54,36.22,35.87,35.55,35.44,33.27,32.18,32.08,31.38,29.35,29.28,29.14,28.92,27.28,26.94,25.45,25.37,24.93,24.74,21.77,21.70,18.41,18.37,14.77,13.46.LC-MS(ESI)[M+Na]+＝531.3.
Example 111
Preparation of the Compound N- (6-fluoro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
The first step of the compound 6-fluoro-1H-indole-2, 3-dione (100 mg,0.606mmol,1.0 eq) was dissolved in concentrated sulfuric acid (2 mL), cooled to 0℃and then added with sodium nitrate (51.47 mg,0.606mmol,1.0 eq) and stirred for 1hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. Slowly adding the reaction solution into ice water (20 mL), quenching, extracting with ethyl acetate (20 mL. Times.2), mixing the organic phases, performing spin-drying under reduced pressure (water pump, 45 ℃) to obtain a crude product, separating and purifying by column chromatography (petroleum ether: ethyl acetate=1:0-2:1), and collecting a new point; the product 6-fluoro-5-nitro-1H-indole-2, 3-dione (111-1) (90 mg,0.343mmol, 56.58%) was obtained as a yellow solid. 1H NMR(400MHz,CD3 OD) δ8.14 (d, j=7.5 hz,1 h), 6.86 (d, j=11.3 hz,1 h).
The second step of adding 6-fluoro-5-nitro-1H-indole-2, 3-dione (111-1) (30 mg,0.143mmol,1.0 eq) to a reaction flask containing tetrahydrofuran (1 mL), cooling to 0deg.C under nitrogen protection, adding borane tetrahydrofuran complex (0.714mL, 0.714mmol,2.3 eq) to room temperature (25deg.C), and stirring for 2hr; TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried under reduced pressure (water pump, 45 ℃) to give crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give the product 6-fluoro-5-nitro-1H-indole (111-2) (15 mg,0.075mmol, 52.49%) as a yellow solid. LC-MS (ESI) [ m+h ] + = 181.1.
The third step, 6-fluoro-5-nitro-1H-indole (111-3) (15 mg,0.083mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), sodium hydride (5.00 mg,0.125mmol,1.7 eq) was added under nitrogen at 0deg.C and stirred for 0.5hr, and methyl iodide (17.73 mg,0.125mmol,1.5 eq) was added and stirring continued for 0.5hr; TLC (petroleum ether: ethyl acetate=3:1) monitored reaction completion. Ethyl acetate (20 mL) was added to the reaction solution, the mixture was washed with water (20 mL. Times.3), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃ C.) to give 6-fluoro-1-methyl-5-nitroindole (111-3) (15 mg,0.062mmol, 74.21%) as a pale yellow solid; LC-MS (ESI) [ m+h ] + = 195.1.
The fourth step compound, 6-fluoro-1-methyl-5-nitroindole (111-4) (15 mg,0.077mmol,1.0 eq), was added to a reaction flask containing methanol (2 mL), and after adding palladium on carbon 10% (20 mg,0.019mmol,0.2 eq), the mixture was stirred at room temperature (25 ℃) under a hydrogen atmosphere for 1hr. TLC (Petroleum ether: ethyl acetate=1:1) monitoring that no raw material remained, had a dominant point of increasing polarity formed, the reaction solution was filtered through celite, the filter cake was washed with methanol (3 mL), and the filtrate was dried under reduced pressure to give the product 6-fluoro-1-methylindol-5-amine (111-5) (15 mg,0.073mmol, 94.64%) as a colorless oily liquid .1H NMR(400MHz,DMSO)δ7.14(d,J＝11.9Hz,1H),7.11(d,J＝2.9Hz,1H),6.85(d,J＝8.7Hz,1H),6.15(d,J＝3.0Hz,1H),4.51(s,2H),3.65(s,3H).
The fifth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (25 mg,0.072mmol,1.1 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (26.72 mg,0.070mmol,1.1 eq) and N, N-diisopropylethylamine (0.023 mL,0.141mmol,2.0 eq) were added, and then stirred at room temperature (25 ℃) for 0.5hr, and 6-fluoro-1-methylindole-5-amine (111-4) (15 mg, 1.09 hr) dissolved in N, N-dimethylformamide (0.5 mL) was added, and then stirred at room temperature (1.0.09 hr) was continued. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, the organic phase was dried over anhydrous sodium sulfate and filtered, the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-2:1) to give the product N- (6-fluoro-1-methylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butyramide (111) (19 mg,0.038mmol, 53.71%) as a white solid .1H NMR(400MHz,MeOD)δ7.69(d,J＝7.4Hz,1H),7.12(d,J＝11.2Hz,1H),7.08(d,J＝3.0Hz,1H),6.34(d,J＝2.8Hz,1H),3.69(s,3H),3.09(dd,J＝10.5,5.5Hz,1H),2.34(s,2H),2.07(ddd,J＝94.5,29.7,23.5Hz,8H),1.63(d,J＝8.7Hz,4H),1.57(s,3H),1.36(dd,J＝13.0,3.0Hz,1H),1.23(s,3H),1.11–0.91(m,3H),0.89(s,3H),0.79(s,3H),0.72(s,3H).13C NMR(101MHz,MeOD)δ174.02,132.49,129.76,129.72,127.20,117.02,117.00,100.60,95.54,95.29,78.40,54.48,54.26,38.53,37.86,36.93,36.23,35.82,33.26,32.09,31.66,28.03,27.25,26.94,26.70,24.45,21.71,21.63,18.39,14.73,13.39.LC-MS(ESI)[M+H]+＝495.3(Ms+1).
Example 112
Preparation of the Compound N' - {4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -1-oxobutylene } -1-methylindole-5-carboxamide
In a first step, the compound 1H-indole-5-carboxylic acid methyl ester (1000 mg, 5.706 mmol,1.0 eq) was dissolved in N, N-dimethylformamide (10 mL), then replaced with nitrogen, cooled to 0℃and then sodium hydrogen (sodium hydride) (273.99 mg,6.850mmol,1.2 eq) was added thereto, stirred for 30min, then methyl iodide (0.93 mL, 8.560 mmol,1.5 eq) was added thereto, stirred overnight at room temperature, TLC (petroleum ether: ethyl acetate=5:1) was detected, water was added thereto to kill when the reaction was completed, then ethyl acetate was extracted (50 mL), the organic phase was washed twice with saturated saline, then dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, and then 1-methylindole-5-carboxylic acid methyl ester (112-1) (1.0 g, 5.92 mmol, 92.59%) was obtained as a white solid by flash chromatography separation and purification (petroleum ether: ethyl acetate=1:0 to 10:1) .1H NMR(400MHz,CDCl3)δ8.39(d,J＝1.5Hz,1H),7.93(dd,J＝8.7,1.6Hz,1H),7.33(d,J＝8.7Hz,1H),7.11(d,J＝3.1Hz,1H),6.58(d,J＝3.1Hz,1H),3.93(s,3H),3.82(s,3H).
In the second step, methyl 1-methylindole-5-carboxylate (112-1) (500 mg,2.643mmol,1.0 eq) was dissolved in methanol (5 mL), then hydrazine hydrate (0.640 mL,13.215mmol,5.0 eq) was added thereto, the mixture was stirred overnight at 70 degrees, the reaction was detected by LC-MS, and after completion of the reaction, the reaction was dried by spin to give a crude product, which was then purified by flash chromatography (dichloromethane: methanol=1:0 to 10:1) to give 1-methylindole-5-carboxamide (112-2) (440 mg,2.33mmol, yield: 87.99%) as a white solid compound. LC-MS (ESI) [ m+h ] + =190.1.
In a third step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108mmol,1.0 eq), 1-methylindole-5-formylhydrazine (112-2) (20.44 mg,0.108mmol,1.0 eq), HATU (41.08 mg,0.108mmol,1.0 eq) was dissolved in N, N-dimethylformamide (5 mL), to which N, N-diisopropylethylamine (0.036 mL,0.216mmol,2.0 eq) and Gibbs reagent (64.37 mg,0.270mmol,2.5 eq) were added, and the mixture was stirred overnight at room temperature. LC-MS detection. After completion of the reaction, water was added thereto to kill, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated sodium chloride, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by preparative plate (dichloromethane: methanol=10:1) to give N' - {4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -1-oxobutylene } -1-methylindole-5-carboxylic acid hydrazide (112-3) (20 mg,0.032mmol, yield ：30.05％).1H NMR(400MHz,CDCl3)δ8.94(s,1H),8.65(s,1H),8.16(s,1H),7.69(d,J＝8.8Hz,1H),7.36(d,J＝8.6Hz,1H),7.14(d,J＝3.0Hz,1H),6.58(d,J＝3.0Hz,1H),3.83(s,3H),3.28–3.13(m,1H),2.32(dd,J＝14.9,7.5Hz,2H),2.13(d,J＝8.3Hz,2H),1.94(d,J＝10.2Hz,2H),1.64(s,6H),1.61(s,3H),1.54(d,J＝4.1Hz,3H),1.42–1.24(m,6H),0.88(d,J＝6.1Hz,12H),0.81(s,3H),0.75(s,3H),0.03(d,J＝2.6Hz,6H).
In a fourth step, the compound N' - {4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1-oxobutylene } -1-methylindole-5-carboxamide (112-3) (20 mg,0.032 mmol) was dissolved in tetrahydrofuran (3 mL), then 1M tetrabutylammonium fluoride (2 mL) was added thereto, and the mixture was warmed to 50℃and stirred overnight, followed by LC-MS detection. After completion of the reaction, water was added thereto to be destroyed, followed by extraction with ethyl acetate (50 mL), followed by washing twice with saturated sodium chloride, drying the organic phase with anhydrous sodium sulfate, filtering, spin-drying to give a crude product, and then separating and purifying (0.1% NH 3/MeCN/H2 O) by preparative chromatography to give N' - {4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -1-oxybutyl } -1-methylindole-5-formylhydrazine (112) (9.6 mg,0.018mmol, 57.37%) as a white solid compound .1H NMR(400MHz,DMSO)δ9.92(d,J＝118.5Hz,2H),8.16(s,1H),7.71(dd,J＝8.6,1.5Hz,1H),7.51(d,J＝8.7Hz,1H),7.42(d,J＝3.1Hz,1H),6.55(d,J＝3.0Hz,1H),4.32(s,1H),3.83(s,3H),3.00(d,J＝7.4Hz,1H),2.29–2.11(m,3H),2.11–1.71(m,6H),1.65(dd,J＝16.6,7.7Hz,3H),1.59(s,3H),1.50(s,4H),1.23(s,3H),0.99(d,J＝11.3Hz,2H),0.91–0.85(m,3H),0.77(s,3H),0.73–0.66(m,3H).13C NMR(101MHz,DMSO)δ171.99,166.87,138.44,133.13,131.57,127.78,127.01,121.07,109.82,102.03,77.39,54.39,54.02,39.37,38.93,38.16,37.06,36.54,33.88,33.52,33.10,32.25,28.87,28.64,27.91,24.69,22.10,21.94,19.87,16.28,14.58,0.57.
LC-MS(ESI)[M+H]+＝520.3.
Example 113
Preparation of Compound 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -1-oxobutylene } amino) -1-methylindole-3-carboxylic acid
Methyl5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1-oxybutyl } amino) -1-methylindole-3-carboxylate (108) (10 mg,0.019mmol,1.0 eq) was dissolved in methanol (5 mL), and sodium hydroxide (15.20 mg,0.380mmol,2.0 eq) was added at room temperature and stirred for 18h. LC-MS detection of the reaction, after completion of the reaction, the methanol was spun off, then quenched with aqueous ammonium chloride, extracted with dichloromethane (50 mL), the organic phase was washed twice with saturated sodium chloride, then dried over sodium sulfate, and spin-dried to give 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1-oxobutylene } amino) -1-methylindole-3-carboxylic acid (113) (8 mg, yield: 78.03%) as a white solid compound .1H NMR(400MHz,DMSO)δ9.80(s,1H),8.25(s,1H),7.94(s,1H),7.53(dd,J＝8.8,1.7Hz,1H),7.40(d,J＝8.8Hz,1H),3.80(s,3H),3.02–2.98(m,1H),2.28(t,J＝7.1Hz,2H),2.19–2.02(m,4H),2.00–1.79(m,5H),1.65(dd,J＝26.4,13.1Hz,4H),1.57(s,3H),1.51(d,J＝6.5Hz,3H),1.37–1.26(m,2H),0.99(dd,J＝12.5,6.4Hz,1H),0.87(s,3H),0.77(s,3H),0.68(s,3H).13C NMR(101MHz,DMSO)δ171.15,170.51,136.62,134.08,133.09,127.02,126.96,115.98,111.84,110.67,106.66,77.39,54.38,54.00,38.92,38.03,37.06,36.74,36.53,33.53,33.44,32.24,28.71,28.64,27.90,24.69,22.08,21.91,19.84,16.26,14.57.LC-MS(ESI)[M+H]+＝521.3
Example 114
Preparation of the Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [5- (1-methylindol-5-yl) -1,3, 4-oxadiazepin-2-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In a first step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108mmol,1.0 eq), 1-methylindole-5-formylhydrazine (112-2) (20.44 mg,0.108mmol,1.0 eq), HATU (41.08 mg,0.108mmol,1.0 eq) was dissolved in N, N-dimethylformamide (5 mL), then N, N-diisopropylethylamine (0.036 mL,0.216mmol,2.0 eq) and Gibbs reagent (64.37 mg,0.270mmol,2.5 eq) were added thereto, and the mixture was stirred overnight at room temperature. LC-MS detection, when the reaction was completed, water was added thereto, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated sodium chloride, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then separated and purified by preparative plate (dichloromethane: methanol=10:1) to give 5- (5- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } -1,3, 4-oxadiazepan-2-yl) -1-methylindole (114-1) (20 mg,0.032mmol, yield ：30.05％)1H NMR(400MHz,CDCl3)δ8.22(s,1H),7.79(d,J＝8.2Hz,1H),7.35(d,J＝8.2Hz,1H),7.12(d,J＝3.0Hz,1H),6.58(d,J＝2.9Hz,1H),3.81(s,3H),3.14(dd,J＝11.2,4.5Hz,1H),2.02(d,J＝7.0Hz,2H),1.75(d,J＝66.1Hz,6H),1.56(d,J＝8.2Hz,4H),1.52(s,3H),1.48–1.28(m,7H),0.99(dd,J＝26.3,13.3Hz,2H),0.88(s,9H),0.82(s,3H),0.72(s,3H),0.68(s,3H),0.02(d,J＝3.1Hz,6H).
In a second step, the compound 5- (5- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1,3, 4-oxadiazepin-2-yl) -1-methylindole (114-1) (20 mg,0.032 mmol) was dissolved in tetrahydrofuran (3 mL), then 1M tetrabutylammonium fluoride (2 mL) was added thereto and the mixture was warmed to 50℃and stirred overnight. LC-MS detection, after completion of the reaction, quench with water, extract with ethyl acetate (50 mL), wash twice with saturated sodium chloride, dry the organic phase with anhydrous sodium sulfate, filter, spin-dry to give crude product, then separate and purify (0.1% NH 3/MeCN/H2 O) by preparative chromatography to give (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [5- (1-methylindol-5-yl) -1,3, 4-oxadiazepine-2-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (114) (11.65 mg,0.023mmol, yield: 69.37%) as white solid compound .1H NMR(400MHz,DMSO)δ8.21(d,J＝1.2Hz,1H),7.80(dd,J＝8.6,1.6Hz,1H),7.65(d,J＝8.6Hz,1H),7.49(d,J＝3.1Hz,1H),6.62(d,J＝2.7Hz,1H),4.34(s,1H),3.87(s,3H),3.04–2.97(m,1H),2.92(t,J＝7.0Hz,2H),2.14(dd,J＝17.6,10.6Hz,3H),1.94(ddd,J＝46.6,22.6,10.4Hz,5H),1.77–1.70(m,1H),1.67–1.61(m,2H),1.58(s,3H),1.51(d,J＝5.3Hz,2H),1.31(dd,J＝14.6,4.8Hz,3H),1.01–0.93(m,2H),0.87(d,J＝6.0Hz,3H),0.77(d,J＝7.5Hz,3H),0.72–0.63(m,3H).13C NMR(101MHz,DMSO)δ166.46,165.54,138.25,132.49,132.07,128.47,127.57,119.79,119.67,114.94,111.17,102.06,77.35,54.31,53.91,38.88,37.89,37.01,36.50,33.48,33.15,32.15,28.58,28.25,25.22,24.99,22.03,21.83,19.80,16.22,14.51.LC-MS(ESI)[M+H]+＝502.3.
Example 115
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) butanamide
Reference example 2, 1-methylpyrrolo [2,3-b ] pyridin-5-amine was changed to 1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-amine to give the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) butanamide (115).1H NMR(400MHz,CDCl3)δ8.13(s,1H),7.77(d,J＝8.8Hz,1H),7.52(s,1H),7.46(d,J＝8.9Hz,1H),4.28(s,3H),3.22(dd,J＝11.4,4.4Hz,1H),2.38(dt,J＝15.8,8.0Hz,2H),2.16(d,J＝8.9Hz,3H),1.98(d,J＝9.5Hz,2H),1.89(d,J＝15.0Hz,2H),1.71(dd,J＝22.0,7.6Hz,6H),1.61(s,3H),1.37(dd,J＝25.2,12.4Hz,2H),1.12–1.00(m,2H),0.96(s,3H),0.86(dd,J＝11.3,5.3Hz,2H),0.80(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ171.61,146.19,134.54,132.35,130.85,127.84,122.20,109.70,109.35,79.12,54.28,53.94,38.83,37.81,37.47,36.95,36.52,34.38,33.58,32.12,29.70,28.48,28.06,27.63,24.29,21.93,21.85,19.68,15.39,14.30.LC-MS(ESI)[M+H]+＝479.4.
Example 116
Preparation of the Compound 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -1-oxobutylene } amino) -1-methylindole-3-carboxamide
In the first step, the compound 1-methyl-5-nitroindole-3-carbonitrile (91-1) (100 mg,0.497mmol,1.0 eq) was dissolved in tetrahydrofuran (1.5 mL) and methanol (1.5 mL), then sodium hydroxide (198.82 mg,4.970mmol,10.0 eq) was added as a solid, and the mixture was warmed to 50℃and stirred overnight. LC-MS detection, when the reaction is completed, spin-drying to obtain crude product, and then separating and purifying by flash chromatography (dichloromethane: methanol=1:0 to 10:1) to obtain 1-methyl-5-nitroindole-3-carboxamide (116-1) (90 mg, yield: 82.61%) as yellow solid compound .1H NMR(400MHz,DMSO)δ9.08(d,J＝2.4Hz,1H),8.22(s,1H),8.11(dd,J＝9.1,2.4Hz,1H),7.73(d,J＝9.1Hz,1H),3.91(s,3H).LC-MS(ESI)[M+H]+＝220.2.
In a second step, the compound 1-methyl-5-nitroindole-3-carboxamide (116-1) (90 mg,0.411mmol,1.0 eq) was dissolved in methanol (5 mL), then palladium (palladium on charcoal) (43.69 mg,0.411mmol,1.0 eq) was added to displace hydrogen, and then stirred at room temperature for 2 hours, and the reaction was detected by LC-MS. When the reaction was completed, it was filtered to give 5-amino-1-methylindole-3-carboxamide (116-2) (36 mg, yield: 37.07) as an oily compound, and the crude product was directly taken as a next step without purification
In a third step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (15 mg,0.043mmol,1.0 eq), 5-amino-1-methylindole-3-carboxamide (116-2) (9.77 mg,0.052mmol,1.2 eq), HATU (24.55 mg,0.065mmol,1.5 eq) was dissolved in N, N-dimethylformamide (5 mL), then N, N-diisopropylethylamine (0.021 mL,0.129mmol,3.0 eq) was added thereto, stirred overnight at room temperature, LC-MS detection was performed, water quenching was added after completion of the reaction, then extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered and spin-dried to give crude product, which was then purified by preparative chromatography (0.1% formic acid/acetonitrile/water) and then lyophilized to give 5- ({ 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -1-oxobutylene } amino) -1-methylindole-3-carboxamide (116) (12.2 mg, yield: 52.9%) as white solid compound .1H NMR(400MHz,DMSO)δ9.77(s,1H),8.28(d,J＝1.6Hz,1H),7.94(s,1H),7.56(dd,J＝8.8,1.7Hz,1H),7.37(d,J＝8.8Hz,1H),4.31(s,1H)3.78(s,3H),3.09–2.96(m,1H),2.27(t,J＝7.1Hz,2H),2.19–1.77(m,7H),1.65(d,J＝13.1Hz,4H),1.57(s,3H),1.51(d,J＝4.9Hz,3H),1.33(d,J＝12.9Hz,1H),1.04–0.92(m,2H),0.87(s,3H),0.78(s,2H),0.77(s,3H),0.68(s,3H).13C NMR(101MHz,DMSO)δ171.08,166.62,133.61,133.11,127.16,126.94,116.05,112.34,110.23,109.69,77.40,54.39,54.00,38.93,38.05,37.06,36.54,33.47,32.24,28.64,27.91,24.73,21.92,19.84,16.27,14.57.LC-MS(ESI)[M+H]+＝520.3.
Example 117
Preparation of the Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [5- (1-methylindol-5-yl) -1,2, 4-oxadiazepin-3-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
The first step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanenitrile (II-2) (100 mg,0.225mmol,1.0 eq) was added to a reaction flask containing 60% aqueous hydroxylamine solution (2 mL) and ethanol (2 mL), and the mixture was stirred for 12 hours after warming to 100 ℃. TLC (dichloromethane: methanol=10:1) monitored the reaction was essentially complete. Dichloromethane (30 mL) was added to the reaction solution, which was washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (dichloromethane: methanol=10:1) to give [ (1Z) -1-amino-4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] buty) hydroxylamine (117-1) (90 mg,0.189mmol, 83%) as a white solid by LC-MS (ESI) [ m+h ] + =477
In a second step, the compound 1-methylindole-5-carboxylic acid (68 mg, 0.348 mmol,2.2 eq) was added to a reaction flask containing anhydrous N, N-dimethylformamide (2 mL), 1-hydroxybenzotriazole (28 mg,0.207mmol,1.2 eq) and N- (propan-2-yl) -1- (propan-2-ylazaylidene) methanimine (44 mg, 0.348 mmol,2.0 eq) were added, and stirring was continued at room temperature (18 ℃) for 0.5hr, followed by addition of [ (1Z) -1-amino-4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydro-butanylidene ] hydroxylamine (117 mg, 1.173 mmol, 1.0.173 mmol). TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 9:1) ({ [ (1Z) -1-amino-4- [ (2 s,4ar,4bs,8ar,10 ar) -2- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrein-8-yl ] butaniden-yl ] amino } oxy) (1-methylindol-5-yl) methanone (117-2) (50 mg,0.079mmol, 46%) as a white solid, which was directly thrown into the next step .1H NMR(400MHz,CDCl3)δ8.39(s,1H),7.94(dd,J＝8.6,1.4Hz,1H),7.34(d,J＝8.7Hz,1H),7.12(d,J＝3.1Hz,1H),6.59(d,J＝3.1Hz,1H),3.83(s,3H),3.18(dd,J＝11.2,4.6Hz,1H),2.34(dd,J＝10.9,4.4Hz,2H),2.17–2.12(m,2H),2.07–1.90(m,7H),1.62(d,J＝7.8Hz,7H),1.29(t,J＝5.9Hz,9H),0.88(d,J＝3.0Hz,11H),0.81(s,3H),0.75(s,3H),0.03(d,J＝2.8Hz,6H).
The third step compound ({ [ (1Z) -1-amino-4- [ (2S, 4aR,4bS,8aR,10 aR) -2- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-8-yl ] butanedino ] amino } oxy) (1-methylindol-5-yl) methanone (117-2) (50 mg,0.079mmol,1 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), a tetrahydrofuran solution of tetrabutylammonium fluoride (0.079 mL,0.079mmol,1eq, 1M) was added and stirred at room temperature for 5 hours, and TLC (petroleum ether: ethyl acetate=1:1) was monitored for the point of reduced polarity of the reaction, and the starting material was completely reacted. Tetrabutylammonium fluoride in tetrahydrofuran (0.79 mL,0.79mmol,10eq, 1M) was added, and the mixture was stirred at 50℃for 16 hours; TLC (dichloromethane: methanol=10:1) monitoring the reaction showed that there was a small amount of starting material remaining, with new spots of increasing polarity; the reaction solution was dried under reduced pressure (water pump, 45 ℃) and added with ethyl acetate (20 mL) and water (20 mL) to dissolve, extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with water (20 mL x 4) and dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (dichloromethane: methanol=10:1) to give the product (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- {3- [5- (1-methylindol-5-yl) -1,2, 4-oxadiazepin-3-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (117) (20 mg,0.04mmol, 50%) as a white solid .1H NMR(400MHz,CDCl3)δ8.45(s,1H),7.98(d,J＝8.7Hz,1H),7.42(d,J＝8.6Hz,1H),7.15(d,J＝3.1Hz,1H),6.61(d,J＝2.9Hz,1H),3.85(s,3H),3.22(dd,J＝11.5,4.6Hz,1H),2.77(dt,J＝14.6,7.2Hz,2H),2.19(dd,J＝16.0,9.7Hz,3H),2.02–1.90(m,4H),1.79–1.73(m,2H),1.68(d,J＝13.2Hz,3H),1.62(s,3H),1.41–1.32(m,2H),1.07–0.99(m,2H),0.96(s,3H),0.89(s,2H),0.80(d,J＝11.1Hz,7H).13C NMR(101MHz,CDCl3)δ176.76,171.09,138.78,132.44,130.64,128.45,127.70,122.11,121.35,115.63,109.78,102.61,79.14,54.29,53.98,38.83,37.81,36.96,36.54,33.57,33.11,32.11,28.59,28.06,27.66,26.23,25.90,21.93,21.85,19.64,15.38,14.30.LC-MS(ESI)[M+H]+＝502.4.
Example 118
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- { [6- (pyrazin-2-yl) hexyl ] amino } methanamide
The first step compound, 2-methylpyrazine (1.00 g,10.625mmol,1.0 eq), was added to a reaction flask containing tetrahydrofuran (20 mL), lithium diisopropylamide (6.375 mL,12.750mmol,1.2 eq) was added at-78deg.C and stirred at-78deg.C for 30min; 1, 5-dibromopentane (1.447 mL,10.625mmol,1.0 eq) was added and stirred at room temperature (20deg.C) for 18h. TLC (petroleum ether: ethyl acetate=2:1) monitored the formation of spots with reduced polarity; the reaction solution was dried under reduced pressure (water pump, 40 ℃) to obtain crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to obtain product 2- (6-bromohexyl) pyrazine (118-1) (1160 mg,4.294mmol, 40.41%) as colorless oily liquid; LC-MS (ESI) [ m+h ] + = 243.05.
The second step compound 2- (6-bromohexyl) pyrazine (118-1) (1.16 g,4.771mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (10 mL), phtK (1.77 g,9.541mmol,2.0 eq) was added, and stirred at room temperature (20deg.C) for 18hr. TLC (petroleum ether: ethyl acetate=1:1) showed dot formation with increased polarity; the reaction solution was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:1), and the new spot (rf=0.55) was collected to give the product 2- [6- (pyrazin-2-yl) hexyl ] isoindole-1, 3-dione (118-2) (1460 mg, 4.247 mmol, 89.03%). LC-MS (ESI) [ m+h ] + = 310.10.
The third step, 2- [6- (pyrazin-2-yl) hexyl ] isoindole-1, 3-dione (118-2) (300 mg,0.970mmol,1.0 eq) was added to a reaction flask containing ethanol (10 mL), N 2H4 (571.10 mg,9.697mmol,10.0 eq) was added, and the mixture was heated to 50deg.C and stirred for 18hr; TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) and then was substituted with oil pump to give 6- (pyrazin-2-yl) hex-1-amine (118-3) (160 mg,0.848mmol, 87.44%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 180.20.
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (118-3) (100 mg,0.223mmol,1.0 eq) was added to a reaction flask containing toluene (2 mL), diphenyl azide phosphate (64.39 mg,0.234mmol,1.05 eq) and triethylamine (0.065 mL, 0.4638 mmol,2.0 eq) were stirred at room temperature (90 ℃) for 1hr, the reaction mixture was cooled to room temperature, and 6- (pyrazin-2-yl) hex-1-amine (39.98 mg,0.223mmol, 1.eq) was further stirred for 18hr; the reaction was monitored by LC-MS for completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- { [6- (pyrazin-2-yl) hexyl ] amino } methanamide (118-4) (65 mg,0.099mmol, 44.30%) as a pale yellow solid. LC-MS (ESI) [ m+h ] + = 626.45.
The fifth step compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- { [6- (pyrazin-2-yl) hexyl ] amino } methanamide (118-4) (65 mg,0.104mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL eq), and tetrabutylammonium fluoride (1.040 mL,1.040mmol,10.0 mmol) was added and then heated to 50℃and stirred for 18hr. TLC (dichloromethane: methanol=10:1) monitored the reaction was essentially complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was prepared on a plate (dichloromethane: methanol=10:1), separated and purified to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- { [6- (pyrazin-2-yl) hexyl ] amino } methanamide (118) (28 mg,0.052mmol, 50.08%) as a white solid .1H NMR(400MHz,CDCl3)δ8.47(d,J＝12.8Hz,2H),8.40(s,1H),4.32(m,2H),3.22(dt,J＝11.0,5.4Hz,2H),3.14(dd,J＝12.9,6.8Hz,2H),3.03(m,1H),2.81(m,2H),2.23(m,2H),1.93(m,2H),1.74(dd,J＝8.8,5.0Hz,3H),1.68(dd,J＝8.6,3.9Hz,2H),1.63(s,3H),1.49(dd,J＝13.4,6.7Hz,2H),1.38(m,4H),1.31(m,2H),1.25(s,2H),1.04(ddd,J＝19.7,10.0,4.8Hz,2H),0.96(s,2H),0.88(m,3H),0.79(d,J＝7.8Hz,6H).13C NMR(101MHz,CDCl3)δ158.24,157.79,144.59,144.02,142.14,129.99,129.64,79.05,54.24,53.87,40.54,39.24,38.82,37.92,36.94,36.50,35.32,33.58,32.20,30.08,29.57,29.26,28.86,28.05,27.62,26.60,21.87,21.82,19.76,15.38,14.28.LC-MS(ESI)[M+H]+＝511.35.
Examples 119&120
Preparation of the Compound (2R) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butanamide
Preparation of the Compound (2S) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butanamide
Compound (127) was resolved using SFC (conditions: column DAICEL CHIRALCEL OJ,250mm30mm I.D, 10 μm; mobile phase: CO2/MeOH [0.2% NH3 (7M Solution in MeOH) ]=70/30; flow rate: 70g/min; wavelength: UV 214nM; temperature: 35 ℃).
(2R) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butanamide (119)1H NMR(400MHz,MeOD)δ7.74(d,J＝1.6Hz,1H),7.29(d,J＝8.8Hz,1H),7.22(dd,J＝8.7,1.8Hz,1H),7.12(d,J＝3.1Hz,1H),6.36(d,J＝3.0Hz,1H),3.76(s,3H),3.13(dd,J＝10.6,5.4Hz,1H),2.51–2.42(m,1H),2.23–2.06(m,3H),1.99(s,2H),1.87(d,J＝19.6Hz,1H),1.82–1.72(m,2H),1.67(d,J＝13.2Hz,2H),1.57(s,3H),1.43–1.32(m,4H),1.27(s,3H),1.19(d,J＝6.9Hz,3H),1.04(ddd,J＝25.1,14.7,8.8Hz,3H),0.92(s,3H),0.83(s,3H),0.81(s,1H),0.76(s,3H).13C NMR(101MHz,MeOD)δ172.21,132.58,129.45,126.89,122.80,121.20,115.87,112.78,108.70,100.35,100.29,78.38,54.45,54.24,42.76,41.39,38.52,37.89,36.90,36.21,33.24,32.03,31.52,29.38,27.23,26.65,26.15,21.70,21.62,18.33,17.43,14.72,13.38.LC-MS(ESI)[M+H]+＝491.4.
(2S) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butanamide (120)1H NMR(400MHz,MeOD)δ7.76(s,1H),7.26(dd,J＝21.2,9.5Hz,2H),7.12(d,J＝2.9Hz,1H),6.35(d,J＝2.9Hz,1H),3.76(s,3H),3.12(dd,J＝10.4,5.3Hz,1H),2.46(dd,J＝13.9,7.0Hz,1H),2.24–2.07(m,3H),2.01(s,2H),1.83(d,J＝12.6Hz,1H),1.70(dd,J＝27.7,16.5Hz,4H),1.59(s,3H),1.36(d,J＝18.8Hz,4H),1.26(s,3H),1.19(d,J＝6.8Hz,3H),1.08–0.97(m,2H),0.91(s,3H),0.88(s,1H),0.81(s,3H),0.74(s,3H).13C NMR(101MHz,MeOD)δ173.00,129.45,128.00,126.80,121.02,115.79,112.67,108.67,103.15,100.28,78.38,54.47,54.26,41.71,38.51,38.00,36.91,36.21,32.13,31.51,29.33,27.24,26.92,26.74,25.26,21.73,21.70,18.34,17.14,17.10,14.68,13.40.LC-MS(ESI)[M+H]+＝491.4.
Example 121
Preparation of the Compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] imidazol-5-yl) amino ] methanamide
Reference example 103, 1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-amine was changed to 1-methylbenzo [ d ] imidazol-5-amine to give the compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } -1- [ (1-methylbenzo [ d ] imidazol-5-yl) amino ] methanamide (121).1H NMR(400MHz,CDCl3)δ8.05(s,1H),7.69(d,J＝1.8Hz,1H),7.45(d,J＝8.7Hz,1H),7.29(dd,J＝8.7,1.9Hz,1H),3.87(s,3H),3.15(m,1H),2.35(m,3H),1.94(dd,J＝29.1,26.8Hz,3H),1.76(dd,J＝18.5,15.3Hz,3H),1.65(s,5H),1.42(dt,J＝19.1,7.2Hz,2H),1.34(s,1H),1.29(m,3H),1.08(m,2H),0.95(s,2H),0.88(d,J＝13.6Hz,2H),0.85(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ157.48,134.74,130.09,129.06,117.19,109.95,109.57,78.37,54.43,54.19,38.52,38.23,37.89,36.90,36.21,33.26,32.16,30.40,29.97,29.36,29.12,27.24,26.92,21.68,21.56,18.51,14.72,13.39.LC-MS(ESI)[M+H]+＝479.30.
Example 122
Preparation of the Compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (3- { [7- (pyrazin-2-yl) heptyl ] amino } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
The first step compound 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (95 mg,0.227mmol,1 eq), 7- (pyrazin-2-yl) hept-1-amine (54-5) (40 mg,0.227mmol,1 eq) was added to a reaction flask containing anhydrous tetrahydrofuran (4 mL), sodium borohydride acetate (53 mg,0.27mmol,1.2 eq) and a drop of glacial acetic acid were stirred at room temperature (25 ℃) for 12 hours, and TLC (dichloromethane: methanol=20:1) monitored for completion of the reaction. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), and the organic phases were combined, washed with water (20 mL x 3) and dried under reduced pressure (water pump, 40 ℃) to give crude product, which was purified by column chromatography (dichloromethane: methanol=1:0 to 15:1) to give the product 2- [7- ({ 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } amino) heptyl ] pyrazine (122-1) (40 mg,0.066mmol, 30%) as a white solid. LC-MS (ESI) [ m+h ] + = 610.06
The second step compound 2- [7- ({ 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } amino) heptyl ] pyrazine (122-1) (40 mg,0.066mmol,1 eq) was added to a reaction flask containing tetrahydrofuran (1 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.810 mL, 0.81mmol, 10eq, 1M) was added and stirred at 50℃for 16 hours; TLC (dichloromethane: methanol=10:1) monitored the reaction was essentially complete. The reaction solution was dried under reduced pressure (water pump, 45 ℃) and added with ethyl acetate (20 mL) and water (20 mL) to dissolve, extracted with ethyl acetate (15 ml×3), the organic phases were combined, washed with water (20 ml×4) and dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by column chromatography (dichloromethane: methanol=10:1) to give the product (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- (3- { [7- (pyrazin-2-yl) heptyl ] amino } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (122) (21.65 mg,0.044mmol, 66.12%) as a white solid .1H NMR(400MHz,CDCl3)δ8.50–8.47(m,1H),8.45(s,1H),8.39(d,J＝2.5Hz,1H),3.21(dd,J＝11.4,4.6Hz,1H),2.84–2.64(m,6H),2.15–1.88(m,7H),1.78–1.63(m,10H),1.59(s,3H),1.55(d,J＝3.7Hz,1H),1.36(s,3H),1.25(s,3H),1.09–1.00(m,2H),0.97(d,J＝7.5Hz,3H),0.92–0.83(m,3H),0.80(d,J＝10.8Hz,7H).13C NMR(101MHz,CDCl3)δ173.68,157.84,144.57,144.04,142.12,126.65,79.08,54.28,53.95,38.82,37.81,36.95,36.52,35.44,33.56,32.15,29.31,29.18,29.13,28.06,27.64,27.22,27.10,27.06,26.54,26.52,21.91,21.84,19.66,15.39,14.30.LC-MS(ESI)[M+H]+＝496.42.
Example 123
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (quinazolin-2-yl) butanamide
Reference example 105 conversion of 5-amino-1- (pyrazin-2-ylmethyl) indole-3-carbonitrile to 2-aminoquinazoline gave the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (quinazolin-2-yl) butanamide (123).1H NMR(400MHz,CDCl3)δ9.28(s,1H),8.42(s,1H),7.87(m,3H),7.53(m,1H),3.22(dd,J＝11.5,4.6Hz,1H),2.88(s,2H),2.22(m,3H),2.00(d,J＝6.8Hz,3H),1.89(m,2H),1.76(m,3H),1.64(s,3H),1.36(dd,J＝14.5,11.1Hz,2H),1.26(s,3H),1.05(m,2H),0.96(s,2H),0.86(m,2H),0.79(d,J＝7.8Hz,6H).13C NMR(101MHz,CDCl3)δ162.29,154.08,150.91,134.89,132.64,127.66,127.40,127.18,126.09,122.10,79.14,54.28,53.96,38.82,37.89,37.40,36.97,36.53,33.58,32.12,29.33,28.62,28.06,27.65,23.83,21.92,21.87,19.69,15.38,14.29.LC-MS(ESI)[M+H]+＝476.25.
Example 124
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {2- [ (4-phenylbutyl) oxy ] ethyl } propionamide
The first step compound 4-phenylbutan-1-ol (1.05 g,6.990mmol,1.0 eq) was added to a reaction flask containing dichloromethane (10 mL), triphenylphosphine (2.02 g,7.689mmol,1.1 eq) was added, and after carbon tetrabromide (2.55 g,7.689mmol,1.1 eq) was stirred at room temperature (20deg.C) for 18hr. TLC (petroleum ether: ethyl acetate=3:1) monitored complete reaction of starting material. The reaction solution was dried under reduced pressure (water pump, 50 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 2:1) to give the product (4-bromobutyl) benzene (124-1) (1.25 g,5.279mmol, 75.52%) as a colorless oil .1H NMR(400MHz,CDCl3)δ7.28(td,J＝7.8,3.9Hz,2H),7.19(m,3H),3.42(t,J＝6.7Hz,2H),2.64(t,J＝7.5Hz,2H),1.90(m,2H),1.78(m,2H).
Sodium hydride (75.08 mg,1.877mmol,2.0 eq) as the second step compound was added to a reaction flask containing anhydrous tetrahydrofuran (2 mL), and ethanolamine (57.32 mg,0.938mmol,1.0 eq) was added dropwise under nitrogen protection and stirred for 0.5hr, while (4-bromobutyl) benzene (124-1) (200 mg,0.938mmol,1.0 eq) (80 ℃ C.) was added and stirred for 18hr. LC-MS monitored reaction was complete. Dilute hydrochloric acid (5 mL) was added to the reaction solution, which was washed with ethyl acetate (10 mL), saturated sodium bicarbonate (10 mL) was added to the aqueous phase, the aqueous phase was extracted with dichloromethane (10 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give the product 2- [ (4-phenylbutyl) oxy ] ethan-1-amine (124-2) (114 mg,0.531mmol, 56% as a colorless oil. LC-MS (ESI) [ m+h ] + = 194.15
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (50 mg,0.111mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), HATU (63.55 mg,0.167mmol,1.5 eq) and N, N-diisopropylethylamine (0.055 mL, 0.332 mmol,3.0 eq) were added and stirring was continued at room temperature (20 ℃) for 0.5hr, followed by 2- [ (4-phenylbutyl) oxy ] ethyl-1-amine (124-2) (25.84 mg,0.134mmol,1.2 eq) and stirring continued for 18hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was isolated and purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {2- [ (4-phenylbutyl) oxy ] ethyl } propionamide (124-3) (37 mg,0.053mmol, 47.89%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 492.40
The fourth step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- {2- [ (4-phenylbutyl) oxy ] ethyl } propionamide (124-3) (37 mg,0.059mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (2 mL), tetrabutylammonium fluoride (0.593 mL,0.593mmol,10.0 eq) was added and the mixture was stirred at 50℃for 18hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by preparative TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- {2- [ (4-phenylbutyl) oxy ] ethyl } propionamide (124) (6 mg,0.01 mmol, 18.46%) as a white solid .1H NMR(400MHz,CDCl3)δ7.29(m,2H),7.18(m,3H),3.73(dd,J＝10.9,6.0Hz,2H),3.50(m,3H),3.25(m,2H),2.65(t,J＝6.7Hz,3H),2.36(m,2H),2.21(m,2H),1.94(m,3H),1.77(dd,J＝13.0,3.1Hz,2H),1.68(m,4H),1.60(s,6H),1.37(m,1H),1.27(m,2H),1.07(ddd,J＝22.8,14.5,10.3Hz,2H),0.99(s,3H),0.88(dd,J＝11.9,9.6Hz,2H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ175.41,141.51,131.99,128.48,128.39,128.29,126.09,79.07,62.87,54.31,53.93,50.15,49.58,49.40,38.83,38.32,36.94,36.51,35.56,33.46,32.27,32.17,28.66,28.53,28.06,27.62,25.08,21.93,21.75,19.53,15.39,14.28.LC-MS(ESI)[M+H]+＝510.35.
Example 125
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylpyrrolio [2,3-c ] pyridin-5-yl) butanamide
The first step compound 5-bromo-1H-pyrrolo [2,3-c ] pyridine (500 mg,2.538mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (10 mL), sodium hydride (73.09 mg,3.045mmol,1.2 eq) was added at 0deg.C, and methyl iodide (0.196 mL,3.045mmol,1.2 eq) was stirred at 20deg.C for 18hr. LC-MS monitored the reaction. Adding 10mL of water to the reaction flask, quenching, extracting with ethyl acetate (20 mL x 3), combining organic phases, washing the organic phases with water (20 mL x 4), drying with anhydrous sodium sulfate, filtering, and drying the filtrate under reduced pressure (water pump, 40 ℃), to obtain a colorless oil; the crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give the product 5-bromo-1-methylpyrrolo [2,3-c ] pyridine (125-1) (227 mg,0.968mmol, 38.14%) as a pale yellow oily solid. LC-MS (ESI) [ m+h ] + =212.15.
The second step compound 5-bromo-1-methylpyrrolo [2,3-c ] pyridine (125-1) (220 mg,1.042mmol,1.0 eq) was added to a reaction flask containing toluene (5 mL), benzophenone imine (283.38 mg,1.564mmol,1.5 eq), tris (dibenzylideneacetone) dipalladium (9.55 mg,0.010mmol,0.01 eq), potassium t-butoxide (116.96 mg,1.042mmol,1.0 eq), BINAP (9.74 mg,0.016mmol,0.015 eq) were added and stirred at 100℃for 18hr. LC-MS monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give 5- [ (diphenylmethylene) amino ] -1-methylpyrrolo [2,3-c ] pyridine (125-2) as a colorless oil, which was used in the next step without purification. LC-MS (ESI) [ m+h ] + = 312.10.
The third step compound 5- [ (diphenylmethylene) amino ] -1-methylpyrrolo [2,3-c ] pyridine (125-2) (324 mg,1.040mmol,1.0 eq) was added to a reaction flask containing dichloromethane (5 mL), HCl (1.040 mL,1.040mmol,1.0 eq) was added and stirred at 20℃for 2hr. LC-MS monitored the reaction. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a colorless oil; the crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:1 to 1:2) to give the product 1-methylpyrrolo [2,3-c ] pyridin-5-amine (125-3) (30 mg,0.183mmol, 17.63%) as a pale yellow solid. LC-MS (ESI) [ m+h ] + = 148.10.
The fourth step compound 44- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (50 mg,0.108mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (5 mL), HATU (24.65 mg,0.065mmol,1.5 eq) and N, N-diisopropylethylamine (0.054 mL,0.324mmol,3.0 eq) were added and stirred at room temperature (20 ℃) for 0.5hr, followed by 1-methylpyrrolio [2,3-c ] pyridin-5-amine (125-3) (15.90 mg,0.108mmol,1.2 eq) and stirred for 18 h. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil. The crude product was isolated and purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-methylpyrazolo [2,3-c ] pyridin-5-yl) butanamide (125-4) (45 mg,0.068mmol, 63.32%) as a pale yellow oil. LC-MS (ESI) [ m+h ] + = 592.40.
The fifth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylpyrrolio [2,3-c ] pyridin-5-yl) butyramide (125-4) (45 mg,0.076mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), tetrabutylammonium fluoride (19.88 mg,0.076mmol,10.0 eq) was added and then stirred at 50℃for 18hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by preparative TLC (dichloromethane: methanol=10:1), to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methylpyrrolio [2,3-c ] pyridin-5-yl) butanamide (125) (25 mg,0.051mmol, 67.34%) as a white solid .1H NMR(400MHz,CDCl3)δ8.55(s,1H),8.48(s,1H),8.37(s,1H),7.24(d,J＝3.0Hz,1H),6.50(d,J＝2.9Hz,1H),3.87(s,3H),3.22(dd,J＝11.4,4.7Hz,1H),2.39(dt,J＝14.9,6.8Hz,3H),2.17(m,3H),1.97(d,J＝10.1Hz,2H),1.88(m,2H),1.76(dt,J＝13.0,3.4Hz,1H),1.69(m,3H),1.61(s,3H),1.38(ddd,J＝26.0,12.8,3.7Hz,2H),1.27(d,J＝12.2Hz,2H),1.06(m,2H),0.97(d,J＝5.7Hz,3H),0.87(m,2H),0.80(d,J＝10.1Hz,6H).13C NMR(101MHz,CDCl3)δ171.34,142.24,136.48,135.02,132.43,131.47,128.22,127.77,104.77,101.37,79.13,54.30,53.98,38.83,37.82,36.97,36.53,33.56,33.25,32.12,29.70,28.49,28.06,27.66,24.31,21.92,21.85,19.65,15.38,14.30.LC-MS(ESI)[M+H]+＝478.30.
Example 126
Preparation of the Compound N- (benzo [2,1-c ] [1,2,5] oxadiazepin-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
Reference example 2, 1-methylpyrrolo [2,3-b ] pyridin-5-amine was changed to benzo [ c ] [1,2,5] oxadiazepine penta-5-amine to give the compound N- (benzo [2,1-c ] [1,2,5] oxadiazepine penta-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] butyramide (126).1H NMR(400MHz,CDCl3)δ8.35(s,1H),7.79(d,J＝9.3Hz,1H),7.42(s,1H),7.30(d,J＝9.5Hz,1H),5.35(s,1H),3.23(dd,J＝11.5,4.6Hz,1H),2.39(m,2H),2.19(m,3H),2.01(s,2H),1.91(s,1H),1.77(m,2H),1.68(d,J＝4.7Hz,2H),1.62(s,3H),1.40(m,2H),1.33(s,2H),1.28(s,3H),1.01(dd,J＝12.1,8.7Hz,2H),0.97(s,2H),0.87(m,2H),0.82(s,3H),0.79(s,2H).13C NMR(101MHz,CDCl3)δ171.96,149.69,147.12,139.18,132.16,128.25,128.05,117.29,101.32,79.10,54.27,53.92,38.83,37.79,36.94,36.52,33.58,32.13,31.43,29.70,29.33,28.06,27.62,23.98,21.93,19.69,15.38,14.30.LC-MS(ESI)[M+H]+＝488.25.
Example 127
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butanamide
The first-step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (100 mg,0.216mmol,1.0 eq) was added to a reaction flask containing methylene chloride (3 mL), oxalyl chloride (0.024 mL,0.281mmol,1.3 eq) and N, N-dimethylformamide (0.002 mL,0.022mmol,0.1 eq) were added, and then stirred at room temperature (25 ℃) for 1hr. TLC (petroleum ether: ethyl acetate=5:1) monitored reaction completion. The reaction solution was slowly added dropwise to a solution of triethylamine (0.060 mL, 0.433 mmol,2.0 eq) in methanol (2 mL,49.376mmol,228 eq) and stirred for 10min, dichloromethane (20 mL) was added, after washing with saturated sodium bicarbonate solution (20 mL x 2), the organic phase was dried under reduced pressure (water pump, 40 ℃) to give crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-19:1) to give methyl 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butyrate (127-1) (100 mg,0.18 mmol, 87.35%) as colorless oil .1H NMR(400MHz,CDCl3)δ3.67(s,3H),3.18(dd,J＝11.2,4.7Hz,1H),2.34–2.23(m,2H),2.15–2.06(m,2H),2.02–1.86(m,3H),1.75–1.61(m,5H),1.59(s,3H),1.56–1.49(m,3H),1.39–1.33(m,1H),1.10–0.96(m,2H),0.93–0.84(m,14H),0.80(s,3H),0.78(d,J＝2.5Hz,1H),0.74(d,J＝7.3Hz,3H),0.03(d,J＝2.8Hz,6H).
The second step compound, methyl 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylphenanthryl ] butyrate (127-1) (60 mg,0.126mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (3 mL), cooled to-78℃under nitrogen protection, and then sodium bistrimethylsilylamino (0.328 mL, 0.178 mmol,6.0 eq) was added and stirred for 1hr, followed by methyl iodide (178.84 mg,1.260mmol,10 eq) and then stirred for 16hr at room temperature (25 ℃). TLC (petroleum ether: ethyl acetate=20:1) monitored reaction completion. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride (20 mL), extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried under reduced pressure (water pump. 40 ℃ C.) to give the crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-24:1) to give the product 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methylbutanoate (127-2) (60 mg,0.110mmol, 87.31%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.64(t,J＝2.1Hz,3H),3.15(dd,J＝11.2,4.6Hz,1H),2.45–2.30(m,1H),2.07(dd,J＝12.4,3.2Hz,1H),1.99(s,2H),1.93(d,J＝14.3Hz,2H),1.66–1.62(m,2H),1.60(d,J＝3.4Hz,2H),1.54(s,3H),1.47(m,1H),1.38–1.33(m,2H),1.14(m,3H),1.06–1.00(m,2H),0.86(s,12H),0.84(s,3H),0.77(s,3H),0.76–0.74(m,2H),0.72(s,3H),0.00(d,J＝2.7Hz,6H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -2-methylbutanoic acid methyl ester (127-2) (50 mg,0.102mmol,1.0 eq) was added to a reaction flask containing methanol (0.5 mL), tetrahydrofuran (0.5 mL) and water (0.5 mL), and after adding lithium hydroxide (85.49 mg,2.037mmol,20 eq), the mixture was stirred at 60℃for 0.5hr. TLC (petroleum ether: ethyl acetate=20:1) monitored reaction completion. The reaction was neutralized to 6-7 with dilute hydrochloric acid (1M), ethyl acetate (20 mL. Times.3) was added to extract, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃ C.) and pulled dry by an oil pump (water bath, 50 ℃ C.) to give the product 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] -2-methylbutanoic acid (127-3) (40 mg,0.076mmol, 74.12%) as a colorless solid
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -2-methylbutanoic acid (127-3) (40 mg,0.084mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), HATU (38.28 mg,0.101mmol,1.2 eq) and diisopropylethylamine (0.028 mL,0.168mmol,2.0 eq) were added, and then stirred at room temperature (25 ℃) for 0.5hr, and then 1-methylindole-5-amine (14.72 mg,0.101mmol,1.2 eq) was added and stirring was continued for 16hr. TLC (petroleum ether: ethyl acetate=3:1) monitored reaction completion. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (20 mL x 3), and dried under reduced pressure (water pump, 45 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-5:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -2-methyl-N- (1-methylindol-5-yl) butyramide (127-4) (25 mg,0.037mmol, 44.33%) as a pale yellow solid .1H NMR(400MHz,MeOD)δ7.76(d,J＝5.2Hz,1H),7.31(d,J＝9.0Hz,1H),7.26(d,J＝2.3Hz,1H),7.14(d,J＝3.0Hz,1H),6.38(d,J＝3.0Hz,1H),3.78(s,3H),3.21(s,1H),2.49(s,1H),2.18(dd,J＝15.5,7.7Hz,3H),2.03(d,J＝5.7Hz,5H),1.69(d,J＝13.3Hz,4H),1.60(d,J＝5.3Hz,6H),1.21(d,J＝6.8Hz,3H),0.90(s,14H),0.84(d,J＝5.9Hz,5H),0.76(d,J＝6.5Hz,3H),0.07–0.04(m,6H).
The fifth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -2-methyl-N- (1-methylindol-5-yl) butanamide (127-4) (25 mg,0.041mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (0.413 mL,0.413mmol,10 eq), the mixture was warmed to 50℃and stirred for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored the reaction was substantially complete. The reaction mixture was dried under reduced pressure (water pump, 45 ℃) to give a brown yellow oil, which was added with ethyl acetate (30 mL), washed with water (20 ml×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product. Separation and purification by preparative-TLC (petroleum ether: ethyl acetate=1:1) gave the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -2-methyl-N- (1-methylindol-5-yl) butanamide (127) (10 mg,0.019mmol, 44.77%) as a white solid .1H NMR(400MHz,MeOD)δ7.76(dd,J＝7.1,1.5Hz,1H),7.31(d,J＝8.8Hz,1H),7.25(ddd,J＝8.7,4.1,1.9Hz,1H),7.14(d,J＝3.1Hz,1H),6.38(d,J＝3.1Hz,1H),3.78(s,3H),3.17–3.11(m,1H),2.53–2.42(m,1H),2.25–2.08(m,3H),2.02(m,2H),1.95–1.82(m,2H),1.82–1.72(m,2H),1.67(m,2H),1.63(m,1H),1.60(d,J＝5.2Hz,3H),1.56(m,1H),1.39(m,2H),1.21(d,J＝6.9Hz,3H),1.12–0.97(m,2H),0.94(d,J＝1.4Hz,3H),0.88(m,1H),0.84(d,J＝6.0Hz,3H),0.82–0.80(m,1H),0.77(d,J＝6.6Hz,3H).13C NMR(101MHz,MeOD)δ173.24,132.60,130.29,129.44,128.51,115.88,115.79,112.79,112.68,108.68,100.29,78.38,54.46,54.24,38.51,37.89,36.90,36.21,33.24,33.07,32.91,31.51,27.23,26.93,23.77,21.69,21.53,21.43,18.33,17.41,14.71,13.40,13.38.LC-MS(ESI)[M+1]+＝491.4.
Example 128
Preparation of the Compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (oxetan-3-yl) indol-5-yl ] butanamide
The first step compound, 5-nitro-1H-indole (300 mg,1.850mmol,1.0 eq), was added to a reaction flask containing N, N-dimethylformamide (6 mL), 3-iodooxetane (336.70 mg,1.850mmol,1.0 eq) and cesium carbonate (1808.44 mg,5.550mmol,3.0 eq) were added, and after heating to 70℃and stirring for 5hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, which was washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-3:1) to give 5-nitro-1- (oxetan-3-yl) indole (128-1) (240 mg,0.990mmol, 53.50%) as a yellow solid .1H NMR(400MHz,CDCl3)δ8.61(d,J＝2.2Hz,1H),8.15(dd,J＝9.1,2.2Hz,1H),7.62(d,J＝3.4Hz,1H),7.51(d,J＝9.1Hz,1H),6.81(d,J＝3.3Hz,1H),5.65–5.57(m,1H),5.23(t,J＝7.4Hz,2H),5.07–5.03(m,2H).
The second step compound, 5-nitro-1- (oxetan-3-yl) indole (128-1) (30 mg,0.137mmol,1.0 eq), was added to a reaction flask containing ethanol (3 mL) and water (1 mL), and after adding iron filings (38.39 mg,0.687mmol,5.0 eq) and ammonium chloride (73.54 mg,1.375mmol,10 eq), the temperature was raised to 80 ℃ and stirred for 4hr.tlc (petroleum ether: ethyl acetate=1:1) to monitor completion of the reaction. The reaction solution was filtered, ethyl acetate (30 mL) was added, washed with water (20 mL x 2) and then dried under reduced pressure (water pump, 45 ℃) to give a crude product which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-1:1) to give the product 1- (oxetan-3-yl) indol-5-amine (128-2) (12 mg,0.051mmol, 37.09%) as a brown solid .1H NMR(400MHz,DMSO)δ7.49(d,J＝3.1Hz,1H),7.23(d,J＝8.7Hz,1H),6.69(d,J＝1.7Hz,1H),6.54(dd,J＝8.6,2.0Hz,1H),6.24(d,J＝2.8Hz,1H),5.66–5.54(m,1H),4.99(t,J＝7.2Hz,2H),4.88(t,J＝6.6Hz,2H),4.59(s,2H).
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (23 mg,0.050mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), HATU (20.79 mg,0.055mmol,1.1 eq) and diisopropylethylamine (0.016 mL,0.099mmol,2.0 eq) were added and stirring was continued at room temperature (25 ℃) for 0.5hr, followed by 1- (oxetan-3-yl) indol-5-amine (128-2) (12.16 mg,0.065mmol,1.3 eq) and stirring continued for 1hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, which was washed with water (20 ml×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product. Purification by preparative-TLC (petroleum ether: ethyl acetate=1:1) gave the product 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] -N- [1- (oxetan-3-yl) indol-5-yl ] butanamide (128-3) (20 mg,0.028mmol, 57.22%) as a white solid .1H NMR(400MHz,CDCl3)δ7.86(s,1H),7.42(d,J＝8.2Hz,2H),7.13(s,1H),6.56(d,J＝3.0Hz,1H),5.59–5.48(m,1H),5.16(t,J＝7.3Hz,2H),5.07(t,J＝6.6Hz,2H),3.18(dd,J＝11.2,4.8Hz,1H),2.35(d,J＝7.6Hz,2H),1.98(dd,J＝65.0,44.8Hz,7H),1.67(s,4H),1.62(s,3H),1.61–1.56(m,2H),1.52(d,J＝17.3Hz,2H),1.43–1.28(m,2H),1.08–0.94(m,2H),0.88(d,J＝5.7Hz,13H),0.81(s,3H),0.75(s,3H),0.03(d,J＝2.6Hz,6H).
The fourth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [1- (oxetan-3-yl) indol-5-yl ] butanamide (128-3) (20 mg,0.032mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a tetrahydrofuran solution of tetrabutylammonium fluoride (0.316 mL,0.316mmol,10 eq) was added thereto and stirred at 50℃for 16hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. The reaction mixture was dried under reduced pressure (water pump, 45 ℃) to give a tan oil, ethyl acetate (30 mL) was added, washed with water (20 mL. Times.4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by preparative-TLC (petroleum ether: ethyl acetate=1:1) to give the product 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [1- (oxetan-3-yl) indol-5-yl ] butyramide (128)(8mg,0.014mmol,44.52％).1H NMR(400MHz,MeOD)δ7.81(s,1H),7.56(d,J＝3.1Hz,1H),7.48(d,J＝8.8Hz,1H),7.29–7.22(m,1H),6.51(d,J＝3.1Hz,1H),5.75–5.62(m,1H),5.17(t,J＝7.3Hz,2H),5.04(t,J＝6.5Hz,2H),3.15(dd,J＝10.7,5.4Hz,1H),2.35(t,J＝5.7Hz,2H),2.20(dd,J＝20.0,10.1Hz,3H),2.02(d,J＝12.1Hz,2H),1.91(s,1H),1.82(d,J＝6.6Hz,1H),1.78(d,J＝13.1Hz,1H),1.68(d,J＝8.8Hz,3H),1.62(s,3H),1.61–1.57(m,1H),1.48–1.39(m,1H),1.34(dd,J＝17.6,10.2Hz,2H),1.14–1.01(m,2H),0.94(s,3H),0.89(d,J＝8.2Hz,1H),0.85(s,3H),0.82(d,J＝10.4Hz,1H),0.77(s,3H).13C NMR(101MHz,CDCl3)δ171.20,132.90,132.49,130.83,129.15,127.73,125.87,116.28,113.13,109.55,102.96,79.12,77.96,77.22,54.30,53.97,50.82,38.83,37.85,37.56,36.97,36.53,33.59,32.13,28.53,28.06,27.65,24.49,21.93,21.87,19.68,15.38,14.30.LC-MS(ESI)[M+1]+＝519.3.
Examples 129&130
Preparation of the Compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -2,4b, 8-tetramethyl-7- [ (methyldioxy-. Lamda.6-thio) amino ] -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propanamide
Preparation of the Compound 3- [ (4 aS,4bR,7R,8aR,10 aR) -2,4b, 8-tetramethyl-7- [ (methyldioxy-. Lamda.6-thio) amino ] -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propanamide
The first-step compound 3- [ (4 bR, 7S) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (79) (340 mg,0.668mmol,1.0 eq) was added to a reaction flask containing dichloromethane (2 mL), pyridinium chlorochromate (288.10 mg,1.337mmol,2.0 eq) and sodium acetate (109.63 mg,1.337mmol,2.0 eq) were added and stirred at room temperature (25 ℃) for 16hr. LC-MS monitoring reaction showed no raw material remaining; the reaction solution was filtered through celite, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give a crude brown oil; the crude product was purified by column chromatography (dichloromethane: methanol=1:0 to 19:1) and Prep-TLC (dichloromethane: methanol=20:1) to give the product 3- [ (4 bR) -2,4b, 8-tetramethyl-7-oxy-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (129-1) (200 mg, 0.35mmol, 53.15%) as a pale yellow oil .1H NMR(400MHz,CDCl3)δ8.53(d,J＝4.9Hz,1H),7.64(td,J＝7.6,1.5Hz,1H),7.16(dd,J＝15.7,7.4Hz,2H),5.49(dd,J＝24.7,11.8Hz,1H),3.23(dd,J＝13.3,6.5Hz,2H),2.81(dd,J＝9.2,6.4Hz,2H),2.68–2.54(m,1H),2.51–2.27(m,3H),2.26–2.08(m,3H),2.06–1.81(m,6H),1.79–1.66(m,4H),1.63(s,3H),1.55–1.42(m,4H),1.36(dt,J＝26.4,13.3Hz,8H),1.05(dd,J＝12.8,6.3Hz,6H),0.98(d,J＝9.6Hz,3H).
The second step compound 3- [ (4 bR) -2,4b, 8-tetramethyl-7-oxy-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (129-1) (200 mg, 0.015 mmol,1.0 eq) was added to a reaction flask containing ethanol (5 mL), hydroxylamine hydrochloride (82.27 mg,1.184mmol,3.0 eq) and sodium acetate (97.12 mg,1.184mmol,3.0 eq) were added, and the mixture was warmed to 50℃and stirred for 3hr; the reaction was monitored by TLC (dichloromethane: methanol=20:1); the reaction solution was dried under reduced pressure (water pump, 45 ℃) to give a white viscous material, dichloromethane (10 mL) and water (20 mL) were added to dissolve, extracted with dichloromethane (15 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, dried under reduced pressure (water pump, 40 ℃) to give a crude product, which was isolated and purified by prep-TLC (dichloromethane: methanol=20:1) to give 3- [ (4 br,7 e) -7- (hydroxyaminomethylene) -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (129-2) (180 mg,0.310mmol, 78.67%) as a colorless oil .1H NMR(400MHz,CDCl3)δ8.59(d,J＝4.0Hz,1H),7.96(s,1H),7.43(s,2H),5.66(s,1H),3.23(dd,J＝13.2,6.4Hz,2H),3.03(s,2H),2.49–2.30(m,2H),2.28–2.14(m,3H),2.02(d,J＝11.9Hz,3H),1.89(d,J＝11.2Hz,2H),1.82(s,3H),1.75–1.67(m,3H),1.63(s,3H),1.54(s,2H),1.52–1.45(m,4H),1.39(dd,J＝15.9,8.4Hz,8H),1.12(d,J＝3.1Hz,3H),1.04(s,3H),0.92(s,3H).
The third step compound 3- [ (4 bR, 7E) -7- (hydroxyamino) -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (129-2) (80 mg,0.153mmol,1.0 eq) was added to a reaction flask containing isopropanol (2 mL), cooled to 0deg.C, then sodium cyanoborohydride (24.09 mg,0.383mmol,2.5 eq), ammonium acetate (29.54 mg,0.383mmol,2.5 eq) and titanium tetrachloride (0.2 mL,0.8 eq) were added under nitrogen protection, and stirred at room temperature (25deg.C) for 2hr. The reaction liquid is monitored by LC-MS to show that no raw material remains; after adjusting the pH to 7-8 with saturated aqueous lithium hydroxide, water (10 mL) was added, extracted with ethyl acetate (20 mL. Times.2), the organic phases were combined, washed with water (20 mL) and dried under reduced pressure (water pump, 45 ℃ C.) to give the crude product, which was purified by prep-TLC (dichloromethane: methanol=10:1) to give the product 3- [ (4 bR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (129-3) (32 mg,0.063mmol, 41.10%) as a yellow oily solid .1H NMR(400MHz,CDCl3)δ8.57(d,J＝5.0Hz,1H),8.32(s,2H),7.86(s,1H),7.34(d,J＝6.6Hz,2H),5.58(d,J＝25.8Hz,1H),3.22(d,J＝6.0Hz,2H),3.02–2.81(m,3H),2.37(d,J＝34.5Hz,2H),2.27–2.08(m,4H),1.98(d,J＝15.4Hz,4H),1.62(s,3H),1.60–1.52(m,3H),1.49(d,J＝6.4Hz,3H),1.35(d,J＝11.2Hz,8H),1.18–1.10(m,3H),1.06(d,J＝21.4Hz,2H),0.97(t,J＝7.1Hz,3H),0.93–0.84(m,3H),0.82(d,J＝7.5Hz,3H).
The fourth step compound 3- [ (4 bR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (129-2) (45 mg,0.089 mmol) was added to a reaction flask containing anhydrous dichloromethane (2 mL), methanesulfonyl chloride (12.18 mg,0.106 mmol) and triethylamine (0.025 mL,0.177 mmol) were added under nitrogen at 0deg.C and then stirred at room temperature (20deg.C) for 1hr; after the completion of the reaction, TLC (dichloromethane: methanol=15:1) monitoring reaction showed that the main point of polarity decrease was generated; dichloromethane (20 mL) was added and washed with water (15 mL x 3) and then dried under reduced pressure (water pump, 40 ℃) to give crude product which was isolated and purified by prep-TLC (dichloromethane: methanol=15:1) and prep-HPLC (column type: xtimate C x30mm mobile phase: acetonitrile-water (0.05% nh 3) 65-80) to give product which was further resolved by SFC (chiral column: chiralpak-IH M mobile phase: CO 2-methanol (0.1% dea)) to give product 3- [ (4 as,4br,7s,8ar,10 ar) -2,4b, 8-tetramethyl-7- [ (methyldioxy- λ6-thio) amino ] -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (pyridin-2-yl) heptyl ] propionamide (129) (12.72 mg,0.021mmol, 24.12%) as a white solid 1H NMR(400MHz,CDCl3)δ8.55(d,J＝4.5Hz,1H),7.72(s,1H),7.26–7.15(m,2H),5.51(s,1H),4.03(d,J＝10.1Hz,1H),3.22(dd,J＝13.1,6.7Hz,2H),3.05–2.93(m,4H),2.85(dd,J＝17.8,10.2Hz,2H),2.44–2.35(m,1H),2.35–2.20(m,2H),2.20–2.07(m,2H),1.99(d,J＝10.7Hz,2H),1.92(m,1H),1.85(dd,J＝16.7,13.1Hz,2H),1.80–1.75(m,2H),1.71(m,2H),1.65(m,2H),1.63(s,3H),1.58(d,J＝3.4Hz,1H),1.51–1.44(m,2H),1.41(m,4H),1.16–1.03(m,2H),1.01(s,3H),0.99–0.90(m,2H),0.86(d,J＝10.8Hz,1H),0.78(d,J＝15.7Hz,6H).13C NMR(101MHz,CDCl3)δ172.83,162.54,139.75,136.58,131.76,128.57,121.09,121.03,77.34,77.23,77.02,76.71,62.74,55.33,53.87,41.72,39.51,38.11,37.85,37.59,36.32,35.61,33.49,32.00,29.76,29.70,29.61,29.20,29.08,28.51,27.78,26.79,25.11,22.43,21.74,19.56,16.19,14.20.LC-MS(ESI)[M+1]+＝586.5.
And 3- [ (4 aS,4bR,7R,8aR,10 aR) -2,4b, 8-tetramethyl-7- [ (methyldioxy-. Lamda.6-thio) amino ] -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [7- (pyridin-2-yl) heptyl ] propanamide (130) (1.98 mg, 0.09 mmol, 3.45%) as a white solid .1H NMR(400MHz,CDCl 3)δ8.53(d,J＝4.8Hz,1H),7.63(t,J＝7.6Hz,1H),7.15(dd,J＝15.7,7.5Hz,2H),5.52(d,J＝6.0Hz,1H),4.06(d,J＝10.1Hz,1H),3.22(dd,J＝13.1,6.6Hz,2H),3.01–2.95(m,4H),2.82–2.76(m,2H),2.22(m,1H),2.16(m,1H),2.05–1.97(m,3H),1.89(m,4H),1.85(m,1H),1.81(m,1H),1.73(m,2H),1.70(m,3H),1.60(s,3H),1.50–1.46(m,2H),1.34(m,4H),1.09(m,1H),1.02(s,3H),0.97(m,1H),0.94(m,1H),0.89(m,2H),0.85(m,1H),0.78(d,J＝5.3Hz,6H).LC-MS(ESI)[M+1]+＝586.5.
Example 131
Preparation of Compound 131 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [5- (6-phenylhexyl) -1,3, 4-oxadiazepin-2-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In the first step 7-phenylheptanoic acid (100 mg, 0.480 mmol) was dissolved in methanol (5 mL), thionyl chloride (86.51 mg,0.727 mmol) was added, heated under reflux for two hours at 50 ℃, the solvent was dried after the reaction was completed, saturated sodium bicarbonate (30 mL) and ethyl acetate (30 mL x 2) were added, the resulting organic phase was dried by spinning to give crude 7-phenylheptanoic acid methyl ester (131), which was directly put into the next reaction. LC-MS: [ m+h ] + =221.1
Second step, 7-phenyl heptanoic acid methyl ester (90 mg,0.409 mmol) was dissolved in methanol (10 mL), then hydrazine hydrate (0.5 mL,10.288 mmol) was added thereto, the mixture was refluxed for 2 hours at 70℃under condensation, filtered and rinsed with methanol, and the crude 7-phenyl heptanoyl hydrazine (131-2) thus obtained was directly spin-dried and put into the next reaction .1H NMR(400MHz,DMSO-d6)δ8.89(s,1H),7.26(m,2H),7.17(m,3H),4.13(s,2H),2.55(m,2H),1.98(t,J＝7.4Hz,2H),1.49(ddd,J＝29.2,14.6,7.2Hz,4H),1.26(m,4H).LC-MS[M+H]+＝221.2
In a third step, 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (71.29 mg, 0.1590 mmol,1.0 eq) and 7-phenylheptanylhydrazine (35 mg, 0.1592 mmol,1.0 eq), HATU (60.40 mg, 0.1592 mmol) were dissolved in DMF (5 mL), then N, N-diisopropylethylamine (0.053 mL,0.318mmol,2.0 eq) was added at room temperature, and primary Gibbs reagent N- (triethylammonium sulfonyl) carbamate (94.64 mg,0.397mmol,2.5 eq) was added, and replaced with nitrogen, and after stirring, the crude product was purified by flash chromatography (5 mL) by direct extraction with water, and saturated aqueous solution of the crude ethyl methyl (4, 6, 8 a-piperidinyl) was obtained by flash chromatography.
The fourth step was to dissolve the compound 2- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -5- (6-phenylhexyl) -1,3, 4-oxadiazepam (131-3) (40 mg,0.063 mmol) in tetrahydrofuran (3.5 mL), then to add tetrabutylammonium fluoride (1.5 mL) at room temperature, warm to 50℃and stir for 18 hours, after completion of the LC-MS detection reaction, water was added thereto, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered and spin-dried to give a crude product, and then subjected to preparative chromatography (formic acid/acetonitrile/water) to give (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [5- (6-phenylhexyl) -1,3, 4-oxadiazepin-2-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (9.5mg,25.5％)1H NMR(400MHz,DMSO-d6)δ7.26(t,J＝7.5Hz,2H),7.22–7.11(m,1H),4.31(dd,J＝13.6,8.8Hz,1H),3.05–2.94(m,1H),2.91–2.65(m,2H),2.62–2.53(m,1H),2.48–2.28(m,1H),2.10(d,J＝11.7Hz,1H),1.93(s,1H),1.80(d,J＝19.2Hz,1H),1.74–1.52(m,4H),1.50(s,2H),1.46–1.17(m,4H),1.00–0.90(m,1H),0.87(s,2H),0.83–0.77(m,1H),0.74(s,2H),0.68(s,2H).13C NMR(101MHz,DMSO-d6)δ166.73,166.57,142.65,131.36,130.05,128.68,128.67,126.07,77.35,54.29,53.85,37.51,37.00,36.50,35.52,33.50,32.05,31.25,28.61,28.51,27.87,26.34,26.02,24.85,24.14,22.00,21.76,19.62,16.25,14.49.LC-MS：[M+H]+＝519.3
Example 132
Preparation of Compound 132- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [ 1-methyl-3- (trifluoromethyl) indol-5-yl ] butanamide
In the first step, 5-nitro-1H-indole (300 mg,1.850 mmol), triethylamine (0.643 mL,4.625 mmol) was dissolved in N, N-dimethylformamide (2 mL), and a solution of iodine (516.54 mg,2.035 mmol) in N, N-dimethylformamide (5 mL) was slowly added dropwise and stirred at room temperature for one hour. After the reaction was completed, 60mL of aqueous ammonia (0.5%) and aqueous sodium thiosulfate (0.1%) were added dropwise, the filter cake was filtered and rinsed with ice water (2×20 mL), and the filter cake was transferred and spin-dried to give crude 3-iodo-5-nitro-1H-indole (132-1) which was directly put into the next reaction. LC-MS: [ m+h ] + =288.9
In a second step, 3-iodo-5-nitro-1H-indole (300 mg,1.042 mmol) was dissolved in N, N-dimethylformamide (2 mL), followed by addition of methyl fluorosulfonyl difluoroacetate (0.663 mL,5.208 mmol) and copper iodide (66.11 mg,0.208 mmol), and heating at 80℃for 3 hours in a sealed tube. After the reaction was completed, 30mL of water was added, extracted with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product, and the crude product was purified by a preparation plate (developer: petroleum ether/ethyl acetate=1/1) to obtain a product of 5-nitro-3- (trifluoromethyl) -1H-indole (132-2)(130mg,0.508mmol,48.81％)1H NMR(400MHz,DMSO-d6)δ12.60(s,1H),8.46(s,1H),8.32(s,1H),8.16(dd,J＝9.1,2.3Hz,1H),7.74(d,J＝9.1Hz,1H).LC-MS：[M-H]-＝229.0
Step three, 5-nitro-3- (trifluoromethyl) -1H-indole (132-2) ((61.15 mg,0.266 mmol) was dissolved in N, N-dimethylformamide (5 mL), then cooled to 0℃under nitrogen protection, sodium hydride (19.13 mg,0.797 mmol) was added to the solution, stirred at 0℃for half an hour, pyrazin-2-ylmethyl methanesulfonate nate (100 mg,0.531 mmol) was added dropwise, warmed to room temperature, stirred for two hours after completion of the reaction, 30mL of water was added, extracted with ethyl acetate (30 mL. 3), the resulting organic phase was dried over anhydrous sodium sulfate and dried to give a crude product, and the crude product was separated by column chromatography (petroleum ether: ethyl acetate=1:1) to give the product 5-nitro-1- (pyrazin-2-ylmethyl) -3- (trifluoromethyl) indole (132-3)(50mg,0.140mmol,52.56％).1H NMR(400MHz,DMSO-d6)δ8.79(d,J＝0.9Hz,1H),8.65(m,1H),8.62(d,J＝2.5Hz,1H),8.33(d,J＝1.2Hz,1H),8.07(s,1H),7.96(s,2H),5.50(s,2H).
In the fourth step, 5-nitro-1- (pyrazin-2-ylmethyl) -3- (trifluoromethyl) indole (132-3) (80 mg,0.248 mmol) was dissolved in methanol (8 mL), palladium on carbon (26.42 mg,0.248 mmol) was added and then stirred at room temperature under hydrogen for 3 hours, after the reaction was completed, the reaction solution was filtered and washed with methanol. The filtrate is dried by spinning to obtain crude 5-nitro-1- (pyrazin-2-ylmethyl) -3- (trifluoromethyl) indole (132-4) which is directly put into the next reaction. LC-MS: [ m+h ] + = 293.1
The fifth step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl 3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (III) (20 mg,0.057 mmol) was added to a reaction flask containing N, N-dimethylformamide (2 mL), HATU (32.73 mg,0.086 mmol) and N, N-diisopropylethylamine (0.028 mL,0.172 mmol) were added and stirred at room temperature (20 ℃) for 0.5hr, then 5-nitro-1- (pyrazin-2-ylmethyl) -3- (trifluoromethyl) indole (16.77 mg,0.057 mmol) was added and stirring was continued for 18hr, and LC-MS showed product formation. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ 1-methyl-3- (trifluoromethyl) indol-5-yl ] butanamide (132) (25 mg,0.042mmol, 74.04%) as a white solid 1H NMR(400MHz,CDCl3)δ8.55(dd,J＝11.6,1.8Hz,2H),8.31(s,1H),7.74(s,1H),7.59(m,2H),7.30(d,J＝8.9Hz,1H),7.23(s,1H),5.45(s,2H),3.22(dd,J＝11.4,4.4Hz,1H),2.34(dt,J＝15.7,8.0Hz,2H),2.16(dd,J＝14.5,6.3Hz,2H),1.94(d,J＝25.1Hz,2H),1.85(dd,J＝14.0,6.6Hz,1H),1.76(dd,J＝13.0,3.3Hz,1H),1.66(m,3H),1.61(d,J＝8.0Hz,3H),1.36(m,2H),1.25(s,3H),1.05(m,2H),0.97(d,J＝6.4Hz,3H),0.90(dd,J＝9.6,5.6Hz,1H),0.85(m,2H),0.81(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ171.30,151.34,144.43,143.07,133.49,132.50,132.41,128.64,127.77,118.11,111.16,110.48,79.12,54.30,53.96,50.30,38.83,37.83,37.48,36.97,36.53,33.58,32.13,29.70,28.49,28.05,27.66,24.38,21.91,21.86,19.67,15.36,14.29.LC-MS:[M+H]+＝623.30
Example 133
Preparation of Compound 133 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (3- {1- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopenta-4-yl } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In the first step, the reaction mixture was stirred at room temperature under nitrogen for 3 hours, and the reaction was monitored by TLC (petroleum ether: ethyl acetate=3:1). After the reaction, adding sodium sulfate decahydrate, quenching, filtering by diatomite, washing 3 times by ethyl acetate, and concentrating in vacuum to obtain a crude product. Purifying the crude product with silica gel column (developing solvent: petroleum ether/ethyl acetate=3/1) to obtain transparent oily (1-methylindol-5-yl) methanol (133-1)(84mg,0.521mmol,91.29％).1H NMR(400MHz,CDCl3)δ7.62(s,1H),7.32(d,J＝8.4Hz,1H),7.24(d,J＝1.2Hz,1H),7.07(d,J＝3.1Hz,1H),6.48(d,J＝3.0Hz,1H),4.77(s,2H),3.80(s,3H).
In a second step, the reaction mixture was stirred at room temperature for 18 hours under nitrogen for TLC (petroleum ether: ethyl acetate=3:1) monitoring, with the addition of reactant (1-methylindol-5-yl) methanol (133-1) (84 mg,0.521 mmol), 1, 8-diazabicyclo [5.4.0] undec-7-ene (158.66 mg,1.042 mmol), diphenyl azide phosphate (215.11 mg,0.782 mmol) to solvent N, N-dimethylformamide (2 mL). After completion of the reaction, water (15 mL) and ethyl acetate (3X 10 mL) were added thereto and extracted. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (developer: petroleum ether/ethyl acetate=10/1) to give 5- (azidomethyl) -1-methylindole (133-2) as a clear oil (37 mg,0.199mmol, 38.13%). LC-MS: [ m+h ] + = 187.1
The third step compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanal (II-3) (100 mg,0.231mmol,1.0 eq) was added to a reaction flask containing methanol (5 mL), dimethyl (1-diazonium-2-oxosubunit propyl) phosphonate (66.59 mg,0.347mmol,1.5 eq) and potassium carbonate (66.85 mg, 0.4632 mmol,2 eq) was added and reacted at room temperature for 1 hour; TLC (petroleum ether: ethyl acetate=20:1) monitored the reaction for completion. Dichloromethane (30 mL) was added to the reaction solution, which was washed with water (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃) to give a crude product which was separated and purified by column chromatography (petroleum ether: ethyl acetate=100:1) to give the product { [ (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- (pent-4-ynyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecaphenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (133-3) (80 mg,0.187mmol, 80.74%) as a white solid .1H NMR(400MHz,CDCl3)δ3.09(dd,J＝11.2,4.7Hz,1H),2.11–2.02(m,5H),1.89–1.82(m,3H),1.61(dd,J＝12.5,8.7Hz,3H),1.53(s,3H),1.46(s,5H),1.41(d,J＝3.4Hz,1H),1.27(dd,J＝13.0,3.6Hz,1H),0.99–0.89(m,2H),0.80(d,J＝5.0Hz,13H),0.72(s,3H),0.67(s,3H),-0.05(d,J＝2.8Hz,6H).
The fourth step gave the product { [ (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (pent-4-ynyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (133-3) (30 mg,0.070mmol,1.0 eq), 5- (azidomethyl) -1-methylindole (133-2) (13 mg,0.070mmol,1 eq), sodium vitamin C (13.86 mg,0.070mmol,1 eq), copper sulfate pentahydrate (17.47 mg,0.070mmol,1 eq) were dissolved in t-butanol (1 mL) and H 2 O (1 mL) and then stirred at room temperature for overnight monitoring (petroleum ether: ethyl acetate=20:1). When the reaction was completed, it was diluted with water, extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered and spin-dried to give 5- [ (4- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylphenanthryl ] propyl } -1,2, 3-triazapentan-1-yl) methyl ] -1-methylindole (133-4) (24.5 mg,0.039mmol, 55.30%) and the crude product was directly used in the next step.
In a fifth step, the compound 5- [ (4- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1,2, 3-triazacyclopenta-1-yl) methyl ] -1-methylindole (133-4) (24.5 mg,0.039mmol,1 eq) was dissolved in tetrahydrofuran (0.5 mL), then tetrabutylammonium fluoride (1M in tetrahydrofuran) (1.5 mL) was added thereto, nitrogen was replaced, and the mixture was warmed to 50℃overnight with stirring, and the LC-MS detection reaction was completed. To this was added water to kill, followed by extraction with ethyl acetate (50 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then subjected to preparative plate separation and purification (petroleum ether: ethyl acetate=2:1) to give (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- (3- {1- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopenta-4-yl } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (133) (18.5 mg,0.036mmol, yield: 92.33%) as a solid compound, and freeze-dried to give a powdery compound.
1H NMR(400MHz,CDCl3)δ7.57(s,1H),7.32(d,J＝8.4Hz,1H),7.17–7.08(m,3H),6.48(d,J＝2.9Hz,1H),5.58(s,2H),3.80(s,3H),3.20(dd,J＝11.5,4.5Hz,1H),2.65(s,2H),2.07–2.00(m,3H),1.89(d,J＝15.0Hz,3H),1.74–1.61(m,7H),1.52(s,3H),1.31(d,J＝9.6Hz,2H),1.03(d,J＝7.6Hz,1H),0.96(d,J＝9.5Hz,3H),0.90–0.80(m,3H),0.77(d,J＝3.4Hz,6H).13C NMR(101MHz,CDCl3)δ136.58,132.80,129.93,129.90,128.62,127.19,121.89,121.13,120.42,109.93,101.16,79.12,54.27,53.95,38.81,37.88,36.95,36.50,33.51,32.99,32.10,29.33,28.76,28.29,28.06,27.65,27.22,25.86,21.88,21.84,19.56,15.37,14.28.LC-MS[M+H]+＝515.
Example 134
Preparation of Compound 134 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [4- (6-phenylhexyl) -1,2, 3-triazacyclopenta-1-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In a first step, 7-phenyloct-1-ol (200 mg,0.97 mmol), pyridine chlorochromate (417.74 mg,1.94 mmol) and solvent dichloromethane (10 mL) were added to a 50mL three-necked flask under nitrogen, and the reaction solution was stirred at room temperature for 1hr. LC-MS monitored the reaction. After completion of the reaction, quenched with water, then extracted with ethyl acetate (15 ml x 3), the organic phase was washed twice with saturated brine (10 ml x 2), then dried over anhydrous sodium sulfate, filtered and spin-dried to give the crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 20:1) to give 7-phenyloctanal (134-1) (100 mg,0.441mmol, yield: 45.45%) as pale yellow oil compound .1H NMR(400MHz,CDCl3)δ9.75(t,J＝1.5Hz,1H),7.26(d,J＝7.1Hz,2H),7.17(t,J＝6.6Hz,3H),2.63–2.58(m,2H),2.41(td,J＝7.3,1.7Hz,2H),1.63(dd,J＝8.1,4.9Hz,4H),1.38–1.33(m,4H).
In a second step, 7-phenyloctanal (134-1) (50 mg,0.245 mmol), (1-diazo-2-oxypropylene) dimethyl phosphonate (94.13 mg,0.490 mmol), potassium carbonate (67.72 mg,0.490 mmol) and solvent methanol (10 mL) were added to a 50mL three-necked flask under nitrogen, and the reaction was stirred at room temperature for 8 hours. LC-MS detection, after the reaction was completed, the reaction solution was diluted with ethyl acetate, and washed with water and saturated brine. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was taken up in silica gel (petroleum ether: ethyl acetate=20:1) and passed through a column, and the sample was collected to give the product oct-7-ynylbenzene (134-2) (25 mg,0.112mmol, yield: 45.90%) as a pale yellow oily compound .1H NMR(400MHz,CDCl3)δ7.29–7.26(m,1H),7.22–7.12(m,1H),2.64–2.58(m,2H),2.18(td,J＝7.0,2.7Hz,1H),1.94(t,J＝2.6Hz,1H),1.62(dd,J＝15.2,7.6Hz,1H),1.55–1.33(m,3H).
In the third step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-azidopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (104-4) (30 mg,0.065 mmol), oct-7-ynylbenzene (134-2) (18.23 mg,0.098 mmol), sodium ascorbate (7.73 mg,0.039 mmol), copper sulfate pentahydrate (3.25 mg,0.013 mmo) were dissolved in t-butanol (2 mL) and H2O (2 mL) and then stirred at room temperature overnight, and after the LC-MS monitoring the reaction was diluted with ethyl acetate and washed with saturated brine. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The product 1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -4- (6-phenylhexyl) -1,2, 3-triazacyclopentanone (134-3) (15 mg,0.022mmol, yield: 33.80%) was obtained as a pale yellow solid compound. The crude product is directly subjected to the next reaction.
The fourth step was to dissolve the compound 1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -4- (6-phenylhexyl) -1,2, 3-triazacyclopenta-ne (134-3) (30 mg,0.045 mmol) in tetrahydrofuran (5 mL), then to which tetrabutylammonium fluoride (1M in tetrahydrofuran) (1.5 mL) was added, nitrogen was replaced, and after the completion of the LC-MS monitoring reaction, water was added thereto for quenching, then extracted with ethyl acetate (30 mL), the organic phase was washed twice with saturated brine, then dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain the crude product, which was then purified by preparative plate separation (petroleum ether) (1.5 mL); ethyl acetate=2:1) to give (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- {3- [4- (6-phenylhexyl) -1,2, 3-triazacyclopent-1-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (134) (15 mg,0.026mmol, yield: 57.44%) as a white solid compound .1H NMR(400MHz,CDCl3)δ7.28(d,J＝8.2Hz,0H),7.24(d,J＝4.3Hz,1H),7.17(t,J＝6.1Hz,3H),4.27(s,2H),3.21(dd,J＝11.5,4.5Hz,0H),2.70(t,J＝7.6Hz,1H),2.63–2.56(m,1H),2.09–1.90(m,2H),1.73–1.57(m,3H),1.54(s,1H),1.41–1.31(m,2H),1.02(s,1H),0.96(s,1H),0.89–0.83(m,1H),0.79(d,J＝8.0Hz,2H).13C NMR(101MHz,CDCl3)δ147.98,142.43,131.09,128.06,128.01,127.95,127.90,125.26,125.20,120.03,78.71,53.93,53.56,49.81,38.48,37.53,36.60,36.16,35.58,33.18,31.73,31.03,29.15,28.82,28.74,28.70,27.71,27.29,25.64,25.32,21.55,21.44,19.18,15.03,13.93.LC-MS[M+1]+＝532.4
Example 135
Preparation of Compound 135N- (3-cyano-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
In a first step, 5-nitro-1H-indole-2, 3-dione (1 g,5.205mmol,1 eq), 5-nitro-1H-indole-2, 3-dione (3.4 mL) and solvent dichloromethane (50 mL) were added to a 50mL three-necked flask, and the reaction was stirred at room temperature for 6 hours. LC-MS monitored the reaction. After the reaction was completed, it was quenched with saturated aqueous sodium bicarbonate, then extracted with ethyl acetate (15 ml×3), the organic phase was washed twice with saturated brine (10 ml×2), then dried over anhydrous sodium sulfate, filtered, and spin-dried to give the crude product, which was then purified by flash chromatography (petroleum ether: ethyl acetate=1:0 to 20:1) to give 3, 3-difluoro-5-nitro-2, 3-dihydro-1H-indol-2-one (135-1) (0.7 g,3.269mmol, yield: 63%) as a yellow solid .1H NMR(400MHz,DMSO-d6)δ11.89(s,1H),8.58(d,J＝1.7Hz,1H),8.43(dd,J＝8.7,2.2Hz,1H),7.20(d,J＝8.7Hz,1H).
In a second step, 3-difluoro-5-nitro-2, 3-dihydro-1H-indol-2-one (400 mg,1.868 mmol) and tetrahydrofuran (6 mL) as solvents were added to a 50mL three-necked flask under nitrogen, and a borane tetrahydrofuran solution (1M) 9.2mL was added dropwise at room temperature and the mixture was allowed to react at 70℃for 5 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate, and washed with water and saturated brine. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was taken up in silica gel (petroleum ether: ethyl acetate=5:1) and passed through a column and the sample was collected to give the product 3-fluoro-5-nitro-1H-indole (135-2) (190 mg,1.055mmol, yield: 56.46%) as a yellow solid. LC-MS: m+h + = 181.1.
In the third step, the compound 3-fluoro-5-nitro-1H-indole (135-2) (500 mg,2.78mmol,1.0 eq) was dissolved in tetrahydrofuran (10 mL), then cooled to 0℃and sodium hydrogen (200 mg,8.33mmol,3.0 eq) was added in portions, reacted for half an hour after warming to room temperature, and then pyrazin-2-ylmethyl methanesulfonate (1044 mg,5.55mmol,2.0 eq) was added. After 1 hour, LC-MS detection was performed, and after completion of the reaction, the reaction mixture was quenched with water, diluted with ethyl acetate, and washed with water and saturated brine. The ethyl acetate layer was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was taken up in silica gel (dichloromethane: methanol/50:1) and passed through a column and the sample was collected to give the product 3-fluoro-5-nitro-1- (pyrazin-2-ylmethyl) indole (135-3) (400 mg,1.47mmol, yield: 52.94%) as a yellow solid compound. LC-MS: m+h + = 273.0.
In a fourth step, 3-difluoro-5-nitro-2, 3-dihydro-1H-indol-2-one (135-3) (400 mg,1.469mmol,1 eq) and methanol (20 mL) as solvents were added to a 50mL single-necked flask, and palladium on carbon (10%) (100 mg) was added at room temperature to replace the reaction mixture with hydrogen atmosphere. LC-MS detection after 1 hour, direct filtration after the reaction, spin drying of the organic phase, and collection of samples gave the product 3-fluoro-1- (pyrazin-2-ylmethyl) indol-5-amine (135-4) (200 mg, 0.8236 mmol, yield: 56.19%) as a yellow solid. LC-MS: m+h + =243.1.
The fifth step was to dissolve the compound 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylhydrophenanthrene ] butanoic acid (III) (50 mg,0.124mmol,1.0 eq), 3-fluoro-1- (pyrazin-2-ylmethyl) indol-5-amine (135-4) (30 mg,0.124mmol,1 eq), HATU (73.64 mg,0.129mmol,1.5 eq) in N, N-dimethylformamide (3 mL), then to which N, N-diisopropylethylamine (0.072 mL,0.215mmol,2.5 mL) was added, stirred at room temperature, LC-MS was detected, after completion of the reaction, water was added, then brine was extracted with ethyl acetate (50 mL), the organic phase was washed twice with saturated aqueous sodium sulfate, the organic phase was dried over a dry sodium sulfate, and the crude petroleum ether was prepared by filtration (dry plate was separated: ethyl acetate=1:1) to give N- (3-cyano-1-methylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (135) (17.67 mg,0.031mmol, yield ：24.91％)1H NMR(400MHz,CDCl3)δ8.56(s,2H),8.19(s,1H),7.80(s,1H),7.31(d,J＝10.3Hz,1H),7.23–7.14(m,2H),7.01(d,J＝2.8Hz,1H),5.36(s,2H),3.22(dd,J＝11.4,4.6Hz,1H),2.41–2.29(m,2H),2.16(t,J＝11.0Hz,3H),2.02–1.89(m,3H),1.70(dd,J＝23.5,9.1Hz,4H),1.62(s,3H),1.55(s,2H),1.42–1.30(m,2H),1.05(dt,J＝17.4,9.6Hz,2H),0.97(d,J＝6.1Hz,3H),0.92–0.85(m,2H),0.80(d,J＝11.9Hz,7H).13C NMR(101MHz,CDCl3)δ171.26,143.99,132.44,130.81,130.45,130.42,127.77,127.74,117.77,115.86,111.46,111.44,109.93,109.15,109.13,79.13,54.29,53.96,50.00,38.84,37.84,37.49,36.96,36.53,33.58,32.13,28.52,28.11,27.65,24.44,21.93,21.86,19.69,15.38,14.31,0.01.LC-MS：[M+H]+＝573.4.
Example 136
Preparation of Compound 136 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [3- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-yl ] butanamide
In the first step, 5-nitro-1H-benzo [ d ] imidazole (850 mg,5.210 mmol) was dissolved in N, N-dimethylformamide (4 mL), then placed under nitrogen, ice-cooled to 0℃and sodium hydride (250.1 mg,10.42 mmol) was added to the solution, stirred at 0℃for half an hour, and pyrazin-2-ylmethyl methanesulfonate (135-1) (120 mg, 0.015 mmol) was added, warmed to room temperature and stirred for two hours. After the reaction was completed, 30mL of water was added, extraction was performed with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product, which was purified by silica gel column (dichloromethane/methanol=10/1) to obtain a mixture, and resolved by isomer preparation to obtain 5-nitro-1- (pyrazin-2-ylmethyl) benzo [ d ] imidazole (136-1)(200mg,0.784mmol,15.04％).1H NMR(400MHz,DMSO-d6)δ8.84(d,J＝1.0Hz,1H),8.69(s,1H),8.57(m,3H),8.17(dd,J＝9.0,2.2Hz,1H),7.81(d,J＝9.0Hz,1H),5.83(s,2H).
In the second step, the compound 5-nitro-1- (pyrazin-2-ylmethyl) benzo [ d ] imidazole (136-1) (200 mg,0.784mmol,1 eq) was dissolved in methanol (5 mL), then palladium on carbon (40 mg,20% wt) was added thereto, hydrogen was replaced, stirring was carried out at room temperature for 30 minutes, after completion of the reaction, filtration was carried out, and then the organic solvent was dried by spinning to give the product 1- (pyrazin-2-ylmethyl) benzo [ d ] imidazole-5-amine (136-2) (36 mg,0.21mmol, yield: 84.21%) as an oily compound. LC-MS: m+h + =226.1.
In a third step, 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylhydrophenanthrene ] butanoic acid (III) (30 mg,0.086mmol,1.0 eq), 1- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-amine (136-2) (23.22 mg,0.103mmol,1.2 eq), HATU (49 mg,0.129mmol,1.5 eq) were dissolved in N, N-dimethylformamide (3 mL), then N, N-diisopropylethylamine (0.036 mL,0.215mmol,2.5 eq) was added thereto under stirring overnight at room temperature, LC-MS was detected, water was added thereto after completion of quenching, then ethyl acetate (50 mL), the organic phase was extracted with brine, saturated aqueous solution was washed twice, and the crude product was purified by a dry sodium sulfate, and the crude product was prepared by filtration, dried (dry phase was separated: ethyl acetate=1:1) to give N- [1- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-yl ] acetamide (2 s,4ar,4bs,8ar,10 ar) -8-ethyl-1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (136) (23 mg,0.04mmol, yield ：46％)1H NMR(400MHz,CDCl3)δ8.50(d,J＝41.9Hz,3H),8.16(s,1H),7.87(s,1H),7.59(d,J＝8.9Hz,1H),7.32(s,2H),5.51(s,2H),3.22(dd,J＝11.5,4.3Hz,1H),2.36(dd,J＝15.6,8.0Hz,3H),2.15(s,4H),1.94(d,J＝23.6Hz,3H),1.75–1.66(m,4H),1.61(s,3H),1.34(d,J＝10.7Hz,1H),1.26(s,1H),1.05(dd,J＝12.7,4.7Hz,2H),0.96(s,3H),0.90–0.84(m,2H),0.79(d,J＝11.3Hz,6H).13C NMR(101MHz,CDCl3)δ204.95,171.35,161.74,146.95,145.49,144.66,144.62,144.58,144.56,143.16,143.15,138.36,127.78,112.30,79.12,54.29,53.96,38.82,37.82,37.49,36.96,36.52,33.58,32.12,28.48,28.05,27.64,24.37,21.92,21.86,19.67,19.65,15.37,14.30.LC-MS：[M+H]+＝556.3.
Example 137
Preparation of Compound 137 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methyl-2-oxo-3H-benzo [ d ] imidazol-5-yl) butanamide
The first step: the reaction 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (III) (20 mg,0.057 mmol), 5-amino-1-methyl-2, 3-dihydro-1H-benzo [ d ] imidazol-2-one (9.36 mg,0.057 mmol), 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate (32.73 mg,0.086 mmol), N, N-diisopropylethylamine (0.028 mL,0.172 mmol) was added to solvent N, N-dimethylformamide (3 mL), and the reaction mixture was stirred at room temperature under nitrogen atmosphere for 16 hours, monitored by LC-MS. After completion of the reaction, water (15 mL) was added thereto, followed by extraction with ethyl acetate (3X 10 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by prep. plate (developer: dichloromethane/methanol=12:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methyl-2-oxo-3H-benzo [ d ] imidazol-5-yl) butanamide as a pale yellow solid (137)(8.49mg,0.017mmol,29.97％).1H NMR(400MHz,DMSO-d6)δ10.76(s,1H),9.74(s,1H),7.52(s,1H),7.07(d,J＝8.3Hz,1H),6.97(d,J＝8.4Hz,1H),4.32(d,J＝3.8Hz,1H),3.24(s,3H),3.00(s,1H),2.28–2.12(m,3H),2.04–1.84(m,5H),1.69–1.47(m,10H),1.34–1.22(m,4H),1.04–0.92(m,2H),0.87(s,3H),0.76(s,3H),0.68(s,3H).13C NMR(101MHz,DMSO-d6)δ171.15,155.03,133.93,133.03,128.55,127.17,126.99,112.09,107.72,101.18,77.37,54.36,53.98,38.92,38.00,37.04,36.70,36.53,32.28,32.22,28.64,27.90,26.83,24.58,22.08,21.88,19.82,16.27,14.57.LC-MS：[M+H]+＝494.3.
Example 138
Preparation of Compound 138 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [1- (5-phenylpentyl) -1,2, 3-triazacyclopentan-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol)
The first step compound (5-bromopentyl) benzene (100 mg,0.440mmol,1.0 eq) was added to a reaction flask containing N, N-dimethylformamide (2 mL), sodium azide (57.24 mg,0.880mmol,2.0 eq) was added, and the mixture was heated to 80℃and stirred for 18hr. The reaction was monitored by TLC (petroleum ether: ethyl acetate=20:1) for completion. After cooling the reaction to room temperature, ethyl acetate (20 mL) was added, washed with water (15 mL x 3), the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was collected and dried under reduced pressure (water pump, 45 ℃) to give the product (5-azidopyl) benzene (138-1) (90 mg,0.380mmol, 86.41%) as a colorless oil, which was used directly in the subsequent reaction.
The second step compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (but-3-ynyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (93-1) (60 mg,0.140mmol,1.0 eq) was added to a reaction flask containing t-butanol (1 mL) and water (1 mL), and (5-azidopyl) benzene (29.13 mg,0.154mmol,1.1 eq), copper sulfate pentahydrate (34.96 mg,0.140mmol,1.0 eq) and ascorbic acid (27.74 mg,0.140mmol,1.0 eq) were added and then stirred at room temperature (25 ℃) for 16h. TLC (petroleum ether: ethyl acetate=5:1) monitored reaction completion. The reaction solution was added with water (20 mL), extracted with ethyl acetate (20 mL x 3), the organic phases were combined, washed with water (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was further purified by column chromatography (petroleum ether: ethyl acetate=1:0-6:1) to give the product 4- {2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } -1- (5-phenylpentyl) -1,2, 3-triazacyclopenta-ne (138-2) (45 mg,0.066mmol, 46.83%) as a white solid .1H NMR(400MHz,CDCl3)δ7.29(t,J＝3.8Hz,3H),7.17(dd,J＝16.7,7.7Hz,3H),4.32(t,J＝7.0Hz,2H),3.24–3.14(m,1H),2.86(s,1H),2.69(d,J＝8.2Hz,1H),2.61(t,J＝7.5Hz,2H),2.43(s,2H),2.21(d,J＝11.2Hz,1H),1.93(dd,J＝15.0,7.4Hz,6H),1.74–1.63(m,6H),1.58(s,3H),1.37(d,J＝8.9Hz,3H),1.02(d,J＝9.0Hz,4H),0.89(s,12H),0.83(d,J＝9.7Hz,4H),0.75(s,3H),0.07–0.02(m,6H).
The third step compound 4- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- (5-phenylpentyl) -1,2, 3-triazacyclopenta-ne (138-2) (45 mg,0.073mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.328 mL, 0.328 mmol,1M,10 eq) was added and stirred at 50℃for 16hr. TLC (petroleum ether: ethyl acetate=2:1) monitored reaction completion. The reaction solution was dried under reduced pressure (water pump, 45 ℃) to give a crude product, ethyl acetate (20 mL) was added, washed with water (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and dried under reduced pressure (water pump, 45 ℃) to give the crude product which was purified by prep-TLC (petroleum ether: ethyl acetate=2:1) to give the product (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- {2- [1- (5-phenylpentyl) -1,2, 3-triazacyclopent-4-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (138) (28.47 mg,0.059mmol, 81.41%) as a white solid ;1H NMR(400MHz,CDCl3)δ7.28(s,2H),7.15(d,J＝7.2Hz,4H),4.33(t,2H),3.23(dd,J＝11.5,4.6Hz,1H),2.88(m,1H),2.71(m,1H),2.61(t,2H),2.43(m,2H),2.22(m,1H),1.99(m,3H),1.91(m,3H),1.81(m,2H),1.76(m,4H),1.58(s,3H),1.37(m,3H),1.09–1.02(m,2H),0.97(s,3H),0.92(d,m,1H),0.87(m,2H),0.82(s,3H),0.79(s,3H).13C NMR(101MHz,CDCl3)δ132.18,128.36,125.84,123.64,79.10,54.31,53.95,50.13,38.84,37.98,36.97,36.53,35.61,33.53,32.18,30.78,30.27,29.06,28.06,27.65,26.06,21.94,21.85,21.73,19.61,15.38,14.31.LCMS：[M+H]+＝504.4.
Example 139
Preparation of Compound 139 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [1- (6-phenylhexyl) -1,2, 3-triazacyclopenta-4-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol)
Reference example 138 (5-bromopentyl) benzene was changed to (6-bromopentyl) benzene to give (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [1- (6-phenylhexyl) -1,2, 3-triazacyclopent-4-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (139) (41.66 mg,0.073mmol, 67.53%) as a white solid ;1H NMR(400MHz,CDCl3)δ7.41(s,1H),7.28(s,2H),7.15(d,J＝7.2Hz,3H),4.37(s,2H),3.21(dd,J＝11.5,4.6Hz,1H),2.86(s,2H),2.60(t,2H),2.12(m,2H),2.07(m,2H),1.95(m,2H),1.92(m,3H),1.76(m,2H),1.67(m,3H),1.63(m,3H),1.61–1.59(m,1H),1.57(s,3H),1.35(m,5H),1.03(m,1H),0.96(s,3H),0.91–0.82(m,3H),0.81(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ142.41,132.81,128.38,128.30,127.26,125.73,120.45,79.11,54.29,53.97,50.20,38.82,37.95,36.97,36.52,35.76,33.54,32.15,31.16,30.31,28.78,28.60,28.42,28.07,27.65,26.37,25.81,21.94,21.86,19.60,15.39,14.30.LC-MS：[M+H]+＝532.3.
Example 140
Preparation of Compound 140- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [3- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-yl ] butanamide
In the first step, 5-nitro-1H-benzo [ d ] imidazole (850 mg,5.21 mmol) was dissolved in N, N-dimethylformamide (4 mL), then placed under nitrogen, ice-cooled to 0℃and sodium hydride (250 mg,10.42 mmol) was added to the solution, stirred at 0℃for half an hour, and pyrazin-2-ylmethyl methanesulfonate (120 mg,0.56 mmol) was added, warmed to room temperature and stirred for two hours. After the reaction was completed, 30mL of water was added, extraction was performed with ethyl acetate (30 mL of 3), the obtained organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product, the crude product was purified by silica gel column (dichloromethane: methanol=10:1) to obtain a mixture, and the mixture was resolved by isomer preparation to obtain 6-nitro-1- (pyrazin-2-ylmethyl) benzo [ d ] imidazole (140-1)(130 mg,0.51 mmol,9.78％).1H NMR(400 MHz,DMSO-d6)δ8.86(d,J＝3.7 Hz,1H),8.74(d,J＝5.1 Hz,1H),8.67(d,J＝2.8 Hz,1H),8.58(m,1H),8.12(ddd,J＝8.8,5.1,2.2 Hz,1H),7.87(dd,J＝8.9,5.0 Hz,1H),5.89(d,J＝4.5 Hz,2H).
In the second step, the compound 5-nitro-1- (pyrazin-2-ylmethyl) benzo [ d ] imidazole (140-1) (200 mg,0.784 mmol,1eq) was dissolved in methanol (5 mL), then palladium on carbon (40 mg,20% wt) was added thereto, hydrogen was replaced, stirring was carried out at room temperature for 30 minutes, after completion of the reaction, filtration was carried out, and then the organic solvent was dried by spinning to give the product 3- (pyrazin-2-ylmethyl) benzo [ d ] imidazole-5-amine (140-2) (36 mg,0.21 mmol, yield: 84.21%) as an oily compound. LC-MS: m+h + =226.
In a third step, 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (III) (30 mg,0.086 mmol,1.0eq), 3- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-amine (140-2) (23.22 mg,0.103 mmol,1.2eq), HATU (49.09 mg,0.129 mmol,1.5eq) were dissolved in N, N-dimethylformamide (3 mL), then N, N-diisopropylethylamine (0.036 mL,0.215 mmol,2.5eq) was added thereto, stirred overnight at room temperature, detected by LC-MS, water was added thereto after completion of the reaction, then ethyl acetate (50 mL) was used for extraction, the organic phase was washed twice with saturated brine, the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product, which was then purified by preparative plate separation (petroleum ether: ethyl acetate=1:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [3- (pyrazin-2-ylmethyl) benzo [ d ] imidazol-5-yl ] butanamide (140) (23 mg,0.04 mmol, yield ：46％)1H NMR(400 MHz,CDCl3)δ8.61–8.40(m,3H),8.19(d,J＝53.3 Hz,2H),7.69(s,1H),7.37(s,1H),6.96(d,J＝7.7 Hz,1H),5.51(s,2H),3.22(dd,J＝11.4,4.5 Hz,1H),2.35(dq,J＝15.3,7.7 Hz,2H),2.15(d,J＝8.7 Hz,3H),1.94(dd,J＝36.1,12.0 Hz,6H),1.76(d,J＝13.1 Hz,1H),1.67(t,J＝8.9 Hz,4H),1.60(s,3H),1.37(dt,J＝12.8,9.9 Hz,1H),1.11–1.00(m,2H),0.96(s,3H),0.88(dd,J＝18.0,9.7 Hz,2H),0.79(d,J＝8.1 Hz,6H).13C NMR(101MHz,CDCl3)δ171.32,150.79,144.56,144.50,143.42,143.12,143.09,134.22,133.95,132.31,127.84,120.59,115.25,101.22,79.09,54.26,53.93,48.03,38.82,37.77,37.63,36.94,36.51,33.57,32.11,28.43,28.05,27.63,24.27,21.91,21.84,19.67,15.38,14.29,-0.01.LC-MS：[M+H]+＝556.
Example 141
Preparation of 2-methylpropan-2-yl Compound 141 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -1-oxobutylene } amino) -1-methylindole-3-carboxylate
In the first step, the compound 1-methyl-5-nitroindole-3-carbonitrile (100 mg,0.50mmol,1.0 eq) was dissolved in ethanol (3 mL) and water (3 mL), then potassium hydroxide (281mg, 5.0mmol,10.0 eq) was added thereto, and stirred for 18 hours at 100℃and TLC (Petroleum ether: ethyl acetate=3:1) was used to monitor the reaction, after the reaction was completed, 1M hydrochloric acid was added thereto to neutralize to weak acidity, then extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to give crude 1-methyl-5-nitroindole-3-carboxylic acid (141-1) (60 mg,0.27mmol, 54%) as a yellow solid compound, which was directly used in the next step.
In a second step, 1-methyl-5-nitroindole-3-carboxylic acid (60 mg, 0.276 mmol) was added to solvent dichloromethane (5 mL), oxalyl chloride (0.069 mL,0.818 mmol) was added dropwise, N, N-dimethylformamide (0.040 mg,0.005 mmol) was stirred at room temperature for 3 hours, after completion of the reaction, the crude product was concentrated in vacuo to give a crude product, potassium tert-butoxide (0.433 mL,0.436 mmol) was added, and the reaction mixture was stirred at 60℃for 3 hours under nitrogen atmosphere, and monitored by TLC (petroleum ether: ethyl acetate=3:1). After the completion of the reaction, ammonium chloride was added to quench the reaction mixture, followed by extraction with water (15 mL) and ethyl acetate (3X 25 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (petroleum ether: ethyl acetate=3:1) to give 1-methyl-5-nitroindole-3-carboxylic acid-2-methylpropan-2-yl ester (141-2) as a pale yellow solid (15 mg,0.054mmol, 19.92%). LC-MS: m+na ] + = 299.1.
In a third step, the compound 1-methyl-5-nitroindole-3-carboxylic acid-2-methylpropan-2-yl ester (141-2) (20 mg,0.072 mmol) was dissolved in methanol (10 mL), pd/C (38.52 mg,0.362 mmol) was then added at room temperature, then hydrogen was replaced, and stirring was carried out at room temperature for 2 hours, and LC-MS monitored. Filtration and spin-drying gave crude 5-amino-1-methylindole-3-carboxylic acid-2-methylpropan-2-yl ester (141-3) (12 mg, 67.3%) which was used directly in the next reaction. LC-MS: [ m+h ] + =247.2.
In a fourth step, 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (IV) (17 mg,0.049mmol,1.0 eq), 5-amino-1-methylindole-3-carboxylic acid-2-methylpropan-2-yl ester (12.01 mg,0.049mmol,1.0 eq) and HATU (27.82 mg,0.073mmol,1.5 eq) were dissolved in DMF (3 mL), then N, N-diisopropylethylamine (0.024 mL,0.146mmol,3.0 eq) was added at room temperature and stirred for 18 hours and LC-MS detected, after completion of the reaction, water quenching was added thereto, followed by extraction with ethyl acetate (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and spin-dried to give a crude product, which was then subjected to preparative chromatography to give 5- ({ 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -1-oxybutyl } amino) -1-methylindole-3-carboxylic acid-2-methylpropan-2-yl ester (141) (8.8 mg, 29.71%) as a white solid compound .1H NMR(400MHz,MeOD)δ8.26(d,J＝1.5Hz,1H),7.82(s,1H),7.41(dt,J＝16.8,5.3Hz,2H),3.83(s,3H),3.14(dd,J＝10.6,5.6Hz,1H),2.37(td,J＝7.1,2.1Hz,2H),2.27–2.12(m,3H),2.07–1.97(m,2H),1.85(ddd,J＝33.3,26.2,9.8Hz,3H),1.73–1.64(m,4H),1.63(s,12H),1.41(dd,J＝12.9,3.3Hz,1H),1.29(s,3H),1.15–1.01(m,2H),0.94(s,3H),0.90(t,J＝6.9Hz,1H),0.84(s,3H),0.76(d,J＝7.0Hz,3H).13C NMR(101MHz,MeOD)δ173.18,165.61,135.97,134.67,132.81,132.48,127.18,126.56,116.71,113.09,109.71,107.82,79.69,78.39,54.47,54.25,38.52,37.83,36.92,36.39,36.23,33.26,32.15,32.10,28.02,27.51,27.25,26.94,24.44,21.72,21.61,18.43,14.72,13.41,-1.45.LC-MS[M+H]+＝577.3.
Example 142
Preparation of Compound 142 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [5- (methylamino) -1H-benzo [ d ] imidazol-2-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In a first step, the reaction product 5-fluoro-2-nitroaniline (10 g,64.05 mmol), methylamine hydrochloride (17.30 g,256.2 mmol), N, N-diisopropylethylamine (31.8 mL,192.2 mmol) and solvent N-methylpyrrolidone (100 mL) were added to a 500mL eggplant bottle to give a yellow solution. The reaction solution was stirred at 100℃for 8 hours. Cooled to room temperature, the reaction mixture was poured into 200mL of ice water, extracted with ethyl acetate (250 ml×2), and the organic layer was washed with saturated brine (250 ml×2), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was isolated and purified by column chromatography (silica gel column, dichloromethane: methanol=100:5) to give 5- (methylamino) -2-nitroaniline (142-1) (9.1 g,54.46mmol, 85.0%) as a yellow solid as LC-MS: [ ms+h ] + =167.8.
In a second step, the reaction product 5- (methylamino) -2-nitroaniline (142-1) (100.0 mg,0.598 mmol), palladium (palladium on charcoal) (10.0 mg,0.094 mmol) and solvent methanol (5 mL) were added to a 100mL eggplant-shaped bottle to give a black suspension. The reaction solution was replaced with nitrogen 3 times and then with hydrogen 3 times. The reaction mixture was stirred under hydrogen (40 psi) at 25℃for 16 hours. The reaction solution was filtered through celite and the mother liquor was concentrated to give 4- (methylamino) benzene-1, 2-diamine (142-2) (70.0 mg,0.459mmol, 78.8%) as a gray solid. LC-MS, [ m+h ] + =138.1.
In a third step, the reactant 4- (methylamino) benzene-1, 2-diamine (142-2) (40.0 mg,0.29 mmol), 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanal (II-3) (100.0 mg,0.22 mmol), acetic acid (0.013 mL,0.22 mmol) and solvent N, N-dimethylformamide (5 mL) were added to a 50mL eggplant-shaped bottle to give a black solution. The reaction solution was stirred at 100℃for 4 hours. The reaction solution was cooled to room temperature, and diluted with ethyl acetate. Washed with water and extracted with ethyl acetate (50 ml x 3). The organic phase was dried over sodium sulfate, filtered and concentrated. The crude product was purified by flash column chromatography (petroleum ether: ethyl acetate=1:1) to give 2- {3- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -5- (methylamino) -1H-benzo [ d ] imidazole (142-4) (65.0 mg,0.083mmol, 37.3%) as a tan solid. LC-MS, [ m+h ] + = 564.7.
In a fourth step, the reaction 2- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propyl } -5- (methylamino) -1H-benzo [ d ] imidazole (142-3) (55.0 mg,0.098 mmol) and solvent hydrogen chloride/1, 4-dioxane (3 mL, 4M) were added to a 25mL eggplant-shaped bottle to give a colorless solution. The reaction solution was stirred at room temperature for 2 hours. LC-MS showed completion of the reaction, concentration of the reaction solution, pH adjustment to 8-9 with sodium bicarbonate, extraction with ethyl acetate (3X 20 mL), combining the organic layers, drying over anhydrous sodium sulfate, filtration concentration and purification by high pressure liquid phase (Waters SunFire C18.6X 150mm 5um,0.05%Formic acid in H2O,ACN) to give (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {3- [5- (methylamino) -1H-benzo [ d ] imidazol-2-yl ] propyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (142)(7.0mg,0.015mmol,15.6％).1H NMR(400MHz,CD3Cl):δ8.59(s,1H),7.33(d,J＝8.7Hz,1H),6.64(d,J＝2.1Hz,1H),6.60-6.55(m,1H),3.24-3.16(m,1H),2.84(s,3H),2.79-2.71(m,3H),2.13-2.02(m,3H),1.96-1.83(m,3H),1.77-1.70(m,2H),1.68-1.58(m,2H),1.36-1.28(m,2H),1.25(s,3H),1.06-0.99(m,2H),0.95(s,3H),0.87-0.82(m,4H),0.80-0.76(m,6H).LC-MS:[M+H]+＝450.4.
Example 143
Preparation of Compound 143 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (2- {1- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopent-4-yl } ethyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
In the first step, the reaction 1-methylindole-5-carboxylic acid (100 mg,0.571 mmol) was dissolved in tetrahydrofuran (3 mL), the system was filled with nitrogen, lithium aluminum hydride (1.142 mL,1.142 mmol) was added dropwise under an ice bath, and the reaction mixture was stirred at room temperature under nitrogen atmosphere for 3 hours, and monitored by TLC (petroleum ether: ethyl acetate=3:1). After the reaction, adding sodium sulfate decahydrate, quenching, filtering by diatomite, washing 3 times by ethyl acetate, and concentrating in vacuum to obtain a crude product. Purifying the crude product with silica gel column (developing solvent: petroleum ether: ethyl acetate=3:1) to obtain transparent oily (1-methylindol-5-yl) methanol (143-1)(84mg,0.521mmol,91.29％).1H NMR(400MHz,CDCl3)δ7.62(s,1H),7.32(d,J＝8.4Hz,1H),7.24(d,J＝1.2Hz,1H),7.07(d,J＝3.1Hz,1H),6.48(d,J＝3.0Hz,1H),4.77(s,2H),3.80(s,3H).
In a second step, the reaction mixture was stirred at room temperature for 18 hours under nitrogen atmosphere for TLC (Petroleum ether: ethyl acetate=10:1) with (1-methylindol-5-yl) methanol (143-1) (84 mg,0.521 mmol), 1, 8-diazabicyclo [5.4.0] undec-7-ene (158.66 mg,1.042 mmol), diphenyl azide phosphate (215.11 mg,0.782 mmol) added to solvent N, N-dimethylformamide (2 mL). After completion of the reaction, water (15 mL) and ethyl acetate (3X 10 mL) were added thereto and extracted. The combined organic phases were dried over anhydrous sodium sulfate and concentrated in vacuo to give the crude product. The crude product was purified by column chromatography over silica gel (petroleum ether: ethyl acetate=10:1) to give 5- (azidomethyl) -1-methylindole (143-2) as a clear oil (37 mg,0.199mmol, 38.13%). LC-MS: m+h + = 187.1.
The third step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanal (I-12) (100 mg,0.231mmol,1.0 eq) was added to a reaction flask containing methanol (10.0 mL), and after adding dimethyl (1-diazo-2-oxopropyl) phosphonate (143-2) (66.59 mg,0.347mmol,1.5 eq), the mixture was stirred at room temperature (25 ℃) for 2hr. TLC (petroleum ether: ethyl acetate=20:1) monitored the reaction to show completion. Ethyl acetate (30 mL) was added to the reaction solution, the organic phase was washed with water (20 mL x 3), and dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-5:1) to give the product { [ (2 s,4ar,4bs,8ar,10 ar) -8- (but-3-ynyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (143-3) (60 mg,0.126mmol, 54.52%) as a colorless oil .1H NMR(400MHz,CDCl3)δ3.18(s,1H),2.32(d,J＝6.9Hz,1H),2.22(dd,J＝12.1,9.5Hz,1H),2.13(d,J＝7.8Hz,2H),1.94(d,J＝9.2Hz,4H),1.72–1.66(m,2H),1.63(s,3H),1.59(s,1H),1.39–1.30(m,1H),1.13–1.03(m,1H),0.91(s,2H),0.89(s,9H),0.88(s,3H),0.80(s,3H),0.76(d,J＝7.2Hz,3H),0.03(d,J＝2.5Hz,6H).
The fourth step compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (but-3-ynyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) monosilane (143-3) (60 mg,0.140mmol,1.0 eq) was added to a reaction flask containing t-butanol (2 mL), water (2 mL), and after adding 5- (azidomethyl) -1-methylindole (26.06 mg,0.140mmol,1.0 eq), copper sulfate pentahydrate (6.99 mg,0.028mmol,0.2 eq) and sodium ascorbate (15.80 mg,0.080mmol,0.57 eq), the mixture was stirred at room temperature (25 ℃) for 18hr. TLC (petroleum ether: ethyl acetate=20:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, which was washed with water (20 mL of 3), and the organic phase was dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-99:1) to give the product 4-methylbenzenesulfonic acid- (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- (2- {1- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopent-4-yl } ethyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrein-2-yl ester (143-4) (45 mg,0.062mmol, 44.19%) as a colorless oil .1H NMR(400MHz,CDCl3)δ7.59(s,1H),7.33(d,J＝8.2Hz,1H),7.13(dd,J＝17.1,5.6Hz,3H),6.49(d,J＝2.9Hz,1H),5.58(s,2H),3.18–3.14(m,1H),2.79(s,1H),2.60(s,1H),2.34(s,2H),2.16–2.10(m,1H),1.90(s,3H),1.75–1.45(m,14H),1.36–1.25(m,3H),0.88(s,9H),0.85(s,3H),0.74(s,3H),0.73(s,3H),0.03(d,J＝3.3Hz,6H).
The fifth step compound 5- [ (4- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1,2, 3-triazacyclopent-1-yl) methyl ] -1-methylindole (143-4) (45 mg,0.073mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding tetrabutylammonium fluoride (191.32 mg,0.732mmol,10.0 eq), the flask was stirred at 50℃for 18hr. TCL (petroleum ether: ethyl acetate=3:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, the organic phase was dried under reduced pressure (water pump, 45 ℃) and washed with water (20 mL), to give a crude product, which was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0-5:1) to give the product (2 s,4ar,4bs,8ar,10 ar) -1, 4a, 7-tetramethyl-8- (2- {1- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopenta-4-yl } ethyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (143) (27 mg,0.052mmol, 71.48%) as a colorless oil .1H NMR(400MHz,CDCl3)δ7.58(s,1H),7.32(d,J＝8.4Hz,1H),7.18–7.08(m,3H),6.49(d,J＝3.0Hz,1H),5.58(d,J＝3.0Hz,2H),3.81(s,3H),3.20(d,J＝7.0Hz,1H),2.85–2.72(m,1H),2.66–2.53(m,1H),2.34(d,J＝6.5Hz,2H),2.22–2.12(m,1H),2.03–1.93(m,1H),1.91(s,1H),1.78–1.72(m,2H),1.72–1.67(m,2H),1.65(d,J＝3.6Hz,2H),1.60(d,J＝3.7Hz,1H),1.56(s,1H),1.48(s,3H),1.33(d,J＝9.4Hz,1H),1.01(td,J＝13.2,3.8Hz,1H),0.95(s,3H),0.84–0.78(m,2H),0.77(s,3H),0.75(s,3H).13C NMR(101MHz,CDCl3)δ129.98,121.96,121.28,109.99,101.23,79.08,73.92,62.31,61.96,58.42,55.15,54.26,53.89,38.81,38.03,37.81,36.94,36.67,36.47,33.39,32.97,32.10,31.49,28.03,27.63,24.92,24.35,21.88,21.70,19.53,15.34,14.23.LC-MS：[M+H]+＝501.3.
Example 144
Preparation of Compound 144N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } -N-methyl-1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] methanamide
The first step compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (I) (300 mg,0.669mmol,1.0 eq) was added to a reaction flask containing toluene (5 mL), diphenyl azide phosphate (189.66 mg,0.669mmol,1.0 eq) and triethylamine (0.195 mL,1.404mmol,2.1 eq) were stirred at 90℃for 1hr, the reaction mixture was cooled to room temperature, and 3mL 6M HCl was further added thereto for 2hr. The reaction was monitored by LC-MS for completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by column chromatography (dichloromethane: methanol=10:1) to give the product (2 s,4ar,4bs,8ar,10 ar) -8- (2-aminoethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (144-1) (150 mg,0.442mmol, 66.10%) as a pale yellow solid. LC-MS: m+h + = 420.30.
The second step compound (2S, 4aR,4bS,8aR,10 aR) -8- (2-aminoethyl) -1, 4a, 7-tetramethyl 1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthrene-2-ol (144-1) (150 mg, 0.356 mmol,1.0 eq) was added to a reaction flask containing methylene chloride (5 mL), and di-tert-butyl dicarbonate (0.099 mL,0.429mmol,1.2 eq) and Et3N (0.099 mL, 0.015 mmol,2.0 eq) were added and stirred at room temperature (20 ℃ C.) for 18hr; the reaction was completed by TLC (petroleum ether: ethyl acetate=1:1). The reaction mixture was taken up in water (15 mL), extracted with dichloromethane (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), affording a colorless oil; the crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:1) to give 2-methylpropan-2-yl ester (144-2) (120 mg,0.208mmol, 58.13%) of product ({ 2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } amino) methane acid as a pale yellow oil LCMS [ m+h ] + = 332.20.
In a third step lithium aluminum hydride (0.460 mL, 0.460 mmol,2.0 eq) was dispersed in tetrahydrofuran (5 mL), a solution of starting material ({ 2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } amino) methanoic acid-2-methylpropan-2-yl ester (144-2) (120 mg,0.231mmol,1.0 eq) in THF (5 mL) was added in ice-bath, and the reaction solution was stirred at 70℃for 18hrs. TLC (dichloromethane: methanol=10:1) showed the reaction was complete. The reaction solution was filtered through celite and the mother liquor was concentrated to give a crude product which was separated and purified by column chromatography (dichloromethane: methanol=10:1) to give the product as a white solid {2- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] ethyl } (meth) amine (144-3) (60 mg,0.124mmol, 53.93%). LC-MS: [ m+h ] + = 434.35
The fourth step {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } (meth) amine (144-3) (60 mg,0.138mmol,1.0 eq) was added to a reaction flask containing dichloromethane (5 mL), N' -carbonyldiimidazole (22.43 mg,0.138mmol,1.0 eq) was added, triethylamine (0.019 mL,0.138mmol,1.0 eq) was stirred at room temperature (20 ℃) for 1hr, and 1-methylbenzo [ d ] [1,2,3] triazacyclopenta-5-amine (20.49 mg,0.138mmol,1.0 eq) was added and stirring was continued for 18hr. The reaction was monitored by LC-MS for completion. The reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was isolated and purified by prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -N-methyl-1- [ (1-methylbenzo [ d ] [1,2,3] triazapentan-5-yl) amino ] methanamide (144-4) (30 mg,0.044mmol, 32.11%) as a pale yellow solid. LC-MS: [ m+h ] + = 608.40
The fifth step compound N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -N-methyl-1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] methanamide (144-4) (30 mg,0.049mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (5 mL), tetrabutylammonium fluoride (0.493 mL,0.493mmol,10.0 eq) was added, and the mixture was stirred at 50℃for 18hr. TLC (dichloromethane: methanol=10:1) monitored reaction was complete. The reaction solution was dried under reduced pressure (water pump, 40 ℃ C.) to give a yellow oil; water (15 mL) was added, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was purified by prep-TLC (dichloromethane: methanol=10:1), to give the product N- {2- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -N-methyl-1- [ (1-methylbenzo [ d ] [1,2,3] triazapentan-5-yl) amino ] methanamide (144) (13 mg,0.026mmol, 53.03%) as a white solid 1H NMR(400MHz,CDCl3)δ7.84(s,1H),7.72(d,J＝8.8Hz,1H),7.43(d,J＝8.8Hz,1H),6.56(s,1H),4.27(s,3H),3.42(m,1H),3.26(m,2H),3.06(m,3H),2.33(m,3H),1.98(d,J＝16.7Hz,2H),1.76(m,2H),1.71(s,3H),1.65(dd,J＝15.7,6.3Hz,2H),1.38(m,2H),1.28(d,J＝3.3Hz,1H),1.25(s,2H),1.06(m,1H),0.98(d,J＝7.2Hz,3H),0.85(dd,J＝15.2,4.9Hz,2H),0.80(d,J＝9.7Hz,6H).13C NMR(101MHz,CDCl3)δ155.51,135.99,130.20,129.94,123.18,109.21,109.09,79.04,54.28,53.94,48.74,38.85,38.76,36.94,36.53,34.96,34.33,33.50,32.34,29.70,28.07,27.78,27.64,21.99,21.76,19.71,15.39,14.28.LC-MS：[M+H]+＝494.30
Example 145
Preparation of Compound 145 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (3- {4- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopent-1-yl } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
The first step was to dissolve the compound 5-bromo-1-methylindole (2000 mg,9.521mmol,1.0 eq) in tetrahydrofuran (50 mL), then replace nitrogen, cool to-78 ℃, then slowly drop butyl lithium (731.87 mg,11.425mmol,1.2 eq), continue stirring for 0.5 hours, then add cuprous iodide (604.27 mg, 1.284 mmol,0.2 eq) and ethylene oxide (2096.92 mg,47.603mmol,5.0 eq), slowly warm to room temperature, stir for 2 hours, LC-MS detects the reaction, after completion of the reaction, quench the reaction with ammonium chloride, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, then filter, spin dry to obtain crude product, separate and purify (petroleum ether: ethyl acetate=1:0 to 2:1) to obtain 2- (1-methylindol-5-yl) ethyl-1-alcohol (145-1) (93 mg, 55.75%) as a solid by flash chromatography .1H NMR(400MHz,CDCl3)δ7.47(d,J＝0.9Hz,1H),7.28(d,J＝8.4Hz,1H),7.09(dd,J＝8.4,1.6Hz,1H),7.04(d,J＝2.5Hz,1H),6.43(d,J＝2.7Hz,1H),3.88(t,J＝6.5Hz,2H),3.78(s,3H),2.97(t,J＝6.5Hz,2H).LC-MS[M+H]+＝176.1.
The second step compound 2- (1-methylindol-5-yl) ethan-1-ol (145-1) (100 mg,0.571mmol,1.0 eq) was added to a reaction flask containing dichloromethane (10.0 mL), followed by addition of dimethyl phthalate (242.05 mg,0.571mmol,2.0 eq) and stirring at room temperature (25 ℃) for 30min; TCL (petroleum ether: ethyl acetate=2:1) monitored reaction completion. LC-MS showed the formation of the product 2- (1-methylindol-5-yl) acetaldehyde (145-2) (32 mg,0.166mmol, 29.13%), LC-MS: [ M+H ] + =174.1.
The third step compound, 2- (1-methylindol-5-yl) acetaldehyde (145-2) (35 mg,0.202mmol,1.0 eq), was added to a reaction flask containing methanol (3.0 mL), followed by addition of dimethyl (1-diazo-2-oxopropyl) phosphonate (58.23 mg,0.303mmol,1.5 eq) and anhydrous potassium carbonate (55.85 mg,0.404mmol,2.0 eq) and stirring at room temperature (25 ℃ C.) for 2hr; TCL (petroleum ether: ethyl acetate=20:1) monitored reaction completion. Ethyl acetate (30 mL) was added to the reaction solution, the organic phase was washed with water (20 mL x 3), and dried under reduced pressure (water pump, 45 ℃) to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0-99:1) to give 1-methyl-5- (prop-2-ynyl) indole (145-3) (18 mg,0.096mmol, 47.38%) as a colorless oil, LC-MS: m+h + =170.1.
The fourth step compound 1-methyl-5- (prop-2-ynyl) indole (145-3) (7.36 mg,0.043mmol,1.0 eq) was added to a reaction flask containing tert-butanol (1 mL) and water (1 mL), and after addition of { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-azidopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] oxy } dimethyl (2-methylpropan-2-yl) silane (104-4) (20 mg,0.043mmol,1.0 eq), copper sulfate pentahydrate (10.86 mg,0.043mmol,1.0 mmol) and sodium ascorbate (8.62 mg,0.043mmol, 1.eq) was stirred at room temperature (20 ℃ C.) for 16hr. TLC (petroleum ether: ethyl acetate=3:1) was monitored completely. The reaction mixture was taken up in water (20 mL), extracted with ethyl acetate (20 mL. Times.3), the organic phases combined, dried under reduced pressure (water pump, 45 ℃ C.) to give crude product, which was isolated and purified by Prep-TLC (petroleum ether: ethyl acetate=3:1) to give the product 5- [ (1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecaphenanthryl ] propyl } -1,2, 3-triazacyclopent-4-yl) methyl ] -1-methylindole (145-4) (24 mg,0.034mmol, 78.95%) as a white solid .1H NMR(400MHz,CDCl3)δ7.51(s,1H),7.28(s,1H),7.13(d,J＝9.0Hz,2H),7.05(d,J＝3.0Hz,1H),6.43(d,J＝3.0Hz,1H),4.23(m,4H),3.78(s,3H),3.16(d,J＝6.5Hz,1H),2.03(m,2H),1.94(m,2H),1.87(m,2H),1.84(m,2H),1.66(m,4H),1.55(m,2H),1.46(m,1H),1.31(m,1H),1.05–0.95(m,3H),0.89(s,9H),0.86(s,3H),0.84–0.80(m,2H),0.78(s,3H),0.76(s,1H),0.74(s,3H),0.03(d,J＝2.6Hz,6H).
The fifth step compound 5- [ (1- {3- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl } -1,2, 3-triazacyclopent-4-yl) methyl ] -1-methylindole (145-4) (24 mg,0.038mmol,1.0 eq) was added to a reaction flask containing tetrahydrofuran (0.5 mL), and after adding a tetrahydrofuran solution of tetrabutylammonium fluoride (0.382 mL,0.382mmol,10 eq), the mixture was stirred at 50℃for 18hr. TLC (petroleum ether: ethyl acetate=1:1) monitored reaction completion. Ethyl acetate (20 mL) was added to the reaction solution, which was washed with water (20 mL. Times.3), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃ C.) to give a crude product, which was purified by Prep-TLC (petroleum ether: ethyl acetate=1:1) to give the product (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- (3- {4- [ (1-methylindol-5-yl) methyl ] -1,2, 3-triazacyclopent-1-yl } propyl) -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (145) (6.06 mg,90.00% purity, 0.01 mmol, 27.77%) as a white solid .1H NMR(400MHz,CDCl3)δ7.52(s,1H),7.29(s,1H),7.16(s,2H),7.06(d,J＝3.0Hz,1H),6.44(d,J＝2.9Hz,1H),4.29(s,2H),4.26(d,J＝6.8Hz,2H),3.79(s,3H),3.21(dd,J＝11.5,4.6Hz,1H),2.04(d,J＝8.2Hz,2H),1.96(m,2H),1.88(m,2H),1.85–1.80(m,1H),1.80–1.70(m,2H),1.66(m,2H),1.59(m,2H),1.50(s,3H),1.37(m,1H),1.32(m,1H),1.02(m,2H),0.96(s,3H),0.81(m,2H),0.78(s,3H),0.78(s,3H).13C NMR(101MHz,CDCl3)δ135.64,131.34,129.79,129.22,129.08,128.72,128.36,122.70,121.47,120.64,109.38,100.67,79.06,54.23,53.87,50.38,38.81,37.80,36.92,36.48,33.49,32.91,32.32,32.05,29.02,28.05,27.63,25.92,21.84,21.76,19.52,15.37,14.27.LC-MS：[M+H]+＝515.3.
Example 146
Preparation of Compound 146N- [1- (difluoromethyl) indol-5-yl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
The first step: in a 100mL eggplant-shaped bottle was added reactant 5-nitro-1H-indole (1.0 g, 6.67 mmol), sodium hydride (0.30 g,7.500 mmol) and solvent N, N-dimethylformamide (10 mL) to give an off-white suspension, which was stirred at room temperature for 10 minutes, [ (2-chloro-2, 2-difluoroacetyl) oxy ] sodium (2.35 g,15.417 mmol) was added and the reaction was heated to 110℃and stirred for 30 minutes. Cooled to room temperature, 100mL of water was added, extracted with ethyl acetate (100 ml×3), dried over anhydrous sodium sulfate, concentrated and then purified by column chromatography (ethyl acetate: petroleum ether=1:5) to give 1- (difluoromethyl) -5-nitroindole as a pale yellow solid (146-1)(450mg,1.909mmol,30.95％).1H NMR(400MHz,DMSO-d6):δ8.69-8.65(m,1H),8.28(s,0.25H),8.22-8.18(m,1H),8.14(s,0.5H),7.99(s,0.25H),7.93-7.87(m,2H).
And a second step of: in a 100mL eggplant-shaped bottle, the reactant 1- (difluoromethyl) -5-nitroindole (146-1) (100 mg,0.471 mmol), palladium (palladium on charcoal) (50.12 mg,0.471 mmol) and solvent methanol (10 mL) were added to give a black suspension, the reaction solution was replaced 3 times with nitrogen and then 3 times with hydrogen. The reaction mixture was stirred under hydrogen balloon pressure at 25 ℃ for 16hrs. Filtration and concentration gave 1- (difluoromethyl) indol-5-amine (146-2) (60 mg, 0.292 mmol, 62.89%) as a pale yellow solid LC-MS: [ M+H ] + =182.8
And a third step of: in a 50mL eggplant-shaped bottle, the reactants 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) and 1- (difluoromethyl) indol-5-amine (146-2) (40.23 mg,0.22 mmol) were dissolved in acetonitrile (1 mL), and then to the reaction solution were added tetramethyl chlorourea hexafluorophosphate (36.38 mg,0.130 mmol) and 1-methylimidazole (0.030 mL,0.378 mmol). The reaction mixture was stirred at 20℃for 2hrs. TLC (petroleum ether: ethyl acetate=2:1) monitoring the reaction showed no starting material remained and a new spot was formed. Adding cold water, for example, to the reaction solution at 0℃to precipitate a white solid, filtering and washing the cake with water to obtain a white solid N- [1- (difluoromethyl) indol-5-yl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) sily ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (146-3) (50 mg,0.072mmol, purity=88%, 66.46%), the crude product being directly used in the next reaction .1H NMR(500MHz,CDCl3):δ8.00-7.91(m,1H),7.57-7.46(m,1H),7.34-7.29(m,1H),7.26-7.22(s,1H),7.16(br s,1H),6.61(d,J＝3.2Hz,1H),3.19(br dd,J＝4.3,11.4Hz,1H),2.46-2.27(m,2H),2.24-2.12(m,2H),2.04-1.96(m,2H),1.96-1.84(m,2H),1.80(br s,1H),1.77-1.67(m,3H),1.64(s,3H),1.58(s,6H),1.46-1.23(m,2H),1.13-0.96(m,2H),0.93(br d,J＝4.7Hz,5H),0.89(br s,3H),0.83(s,4H),0.80-0.72(m,4H),0.05(d,J＝3.4Hz,6H).LCMS:[M+H]+＝627.4
Fourth step: n- [1- (difluoromethyl) indol-5-yl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (146-3) (80 mg,0.134 mmol) was dissolved in methanol (4 ml) at 20℃and (7, 7-dimethyl-2-oxobicyclo [2.2.1] hept-1-yl) methanesulfonic acid (33.35 mg,0.144 mmol) was added. The reaction mixture was stirred at 30℃for 2h. LC-MS showed the reaction was complete. The mixture is diluted with water and filtered under ice water conditions to obtain a solid, and N- [1- (difluoromethyl) indol-5-yl ] -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide is prepared by high pressure (146)1H NMR(400MHz,CDCl3):δ7.95(s,1H),7.50(d,J＝8.7Hz,1H),7.37(s,1H),7.39-7.29(m,1H),7.18-7.05(m,1H),6.60(d,J＝3.3Hz,1H),3.23(dd,J＝4.6,11.4Hz,1H),2.44-2.29(m,2H),2.21-2.13(m,2H),2.08-1.96(m,2H),1.95-1.84(m,2H),1.81-1.70(m,3H),1.67-1.61(m,4H),1.54(br s,4H),1.46-1.33(m,1H),1.15-1.02(m,2H),1.01-0.95(m,2H),0.99-0.91(m,4H),0.81(d,J＝12.3Hz,7H).LCMS:[M+H]+＝513.3.
Example 147
Preparation of Compound 147 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (quinazolin-7-yl) butanamide
Reference example 2 conversion of 1-methylpyrrolo [2,3-b ] pyridin-5-amine to quinazolin-7-amine gives the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (quinazolin-7-yl) butanamide (147).1H NMR(400MHz,MeOD)δ9.51-9.31(m,1H),9.15(br,1H),8.49(d,1H),8.07(d,J＝8.8Hz,1H),7.87(dd,1H),3.15(dd,1H),2.49-2.43(m,2H),2.26-2.11(m,3H),2.08-1.95(m,2H),1.91–1.81(m,2H),1.79–1.65(m,5H),1.63(s,4H),1.41(m,1H),1.14–0.99(m,2H),0.94(s,3H),0.91-0.87(m,1H),0.87-0.85(m,1H),0.85(s,3H),0.83(d,1H),0.77(s,3H).LC-MS：[M+H]+＝476.3.
Example 148
Preparation of Compound 148 (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [5- (6-phenylhexyl) -1,2, 4-oxadiazepin-3-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol
Reference example 117 conversion of 1-methylindole-5-carboxylic acid to 7-phenylheptanoic acid gave the compound (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {2- [5- (6-phenylhexyl) -1,2, 4-oxadiazepin-3-yl ] ethyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-ol (148).1H NMR(400MHz,CHLOROFORM-d)δ7.41-7.29(m,2H),7.26-7.15(m,3H),3.25(br dd,J＝4.6,11.5Hz,1H),2.95-2.74(m,3H),2.72-2.60(m,3H),2.58-2.44(m,2H),2.24(br dd,J＝2.9,12.2Hz,1H),2.10-1.96(m,2H),1.89-1.71(m,6H),1.66-1.55(m,9H),1.49-1.37(m,5H),18-0.97(m,4H),0.91-0.77(m,8H).LC-MS[M+H]+＝519.5.
Example 149
Preparation of Compound 149 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (4, 4-trifluorobutyl) propanamide
Reference example 36, 1-methylindol-5-amine was replaced with 4, 4-trifluorobutylamine, obtaining the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (4, 4-trifluorobutyl) propionamide (149).1H NMR(400MHz,MeOD)δ3.28-3.23(m,1H),3.22-3.12(m,2H),2.54-2.43(m,1H),2.34-2.11(m,6H),2.10-1.90(m,3H),1.89-1.54(m,11H),1.51-1.38(m,1H),1.11-1.00(m,2H),0.96(s,3H),0.90(br d,J＝3.5Hz,1H),0.86(s,3H),0.84(br s,1H),0.79(s,3H).LC-MS[M+H]+＝444.3.
Example 154
Example 154 preparation of N- [7- (3-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
In the first step m-fluorobromobenzene (118.95 mg,0.680 mmol), [ (7-bromoheptyl) amino ] methanoic acid-2-methylpropan-2-yl ester (100 mg,0.340 mmol), sodium iodide (12.74 mg,0.085 mmol), zinc powder (44.44 mg,0.680 mmol), 2-amidinopyridine hydrochloride (5.36 mg,0.034 mmol), trifluoroacetic acid (3.876 mg,0.034 mmol), nickel (II) ethylene glycol dimethyl ether complex (7.47 mg,0.034 mmol) were added sequentially to DMA (N, N-dimethylformamide) (5 mL), the reaction mixture was stirred 16h at 60 ℃ to room temperature, quenched with water (5 mL), extracted with ethyl acetate (5 mL 3), the organic phases combined and washed with saturated brine, dried over anhydrous sodium sulfate to give the filtrate, followed by concentration under reduced pressure to give the crude product, which was purified by scraping on a large plate (ea=5:7.034 mmol) to give [ 7-3- (3-phenyl) propan-2-yl) methyl-2-yl ester (5 mL) (colorless 1.70 mmol). LC-MS [ m+h ] + =210.1.
In the second step { [7- (3-fluorophenyl) heptyl ] amino } methanoic acid-2-methylpropan-2-yl ester (154-1) (60 mg,0.194 mmol) was added to a mixed solution of dichloromethane (0.5 mL) and trifluoroacetic acid (0.1 mL), and stirring was carried out at 20℃for 1h.LCMS showed complete reaction, and the reaction solution was spin-dried to give 7- (3-fluorophenyl) hept-1-amine (154-2) (40 mg,0.191mmol, 98.45%) as a white solid, which was directly used in the next step. LC-MS [ m+h ] + =210.1.
In a third step, 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (80 mg,0.239 mmol), 7- (3-fluorophenyl) hept-1-amine (154-2) (40 mg,0.191 mmol) was added to a reaction flask containing anhydrous acetonitrile (1 mL), N-methylimidazole (68.73 mg,0.837 mmol) was added thereto, the mixture was stirred at 20℃for 1hr, N, N, N ', N' -tetramethyl chloroformidine hexafluorophosphate (100.66 mg, 0.351 mmol) was added, then stirring at 20deg.C for 2hr, LCMS detecting completion of the reaction, adding water to quench, extracting with ethyl acetate (50 mL), drying the organic phase with anhydrous sodium sulfate, filtering, spin-drying to obtain crude product, and subjecting to preparative chromatography to obtain N- [7- (3-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (154) (26.49 mg,0.050mmol, 21.07%) as white solid compound .1H NMR(400MHz,CDCl3)δ7.26-7.17(m,1H),7.00-6.79(m,3H),5.53(br s,1H),3.31-3.12(m,3H),2.60(t,J＝7.7Hz,2H),2.48-2.21(m,4H),2.19-2.09(m,2H),1.99-1.89(m,2H),1.79-1.68(m,3H),1.66-1.56(m,6H),1.52-1.44(m,2H),1.41-1.27(m,7H),1.10-0.94(m,5H),0.93-0.75(m,9H).LC-MS[M+H]+＝526.4.
Example 155
Example 155 preparation of N- [7- (3-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
In a first step p-bromofluorobenzene (920.67 mg,5.261 mmol), 7-bromoheptonitrile (1000 mg,5.261 mmol), sodium iodide (197.14 mg,1.315 mmol), zinc powder (687.92 mg,10.522 mmol), 2-amidinopyridine hydrochloride (82.91 mg,0.526 mmol), trifluoroacetic acid (59.99 mg,0.526 mmol), nickel (II) chloride glycol dimethyl ether complex (115.59 mg,0.526 mmol) were added successively to DMA (N, N-dimethylacetamide) (15 mL), the reaction mixture was stirred at 60℃for 16hr, cooled to room temperature, quenched with water (5 mL), extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate to give a filtrate, and then concentrated under reduced pressure to give a crude product which was separated by column chromatography (petroleum ether: ethyl acetate=20:1) to give 7- (4-fluorophenyl) heptonitrile (155-1) (15 mg, 2.67 mmol, 41.192%) as a colorless oil .1H NMR(400MHz,CDCl3)δ7.11(dd,J＝8.5,5.5Hz,2H),7.03-6.91(m,2H),2.58(t,J＝7.7Hz,2H),2.32(t,J＝7.0Hz,2H),1.67-1.56(m,4H),1.53-1.42(m,2H),1.39-1.31(m,2H).
In the second step, 7- (4-fluorophenyl) heptanenitrile (155-1) (200 mg,0.974 mmol), raney nickel (212.88 mg,0.974 mmol) was added to methanol (10 mL) at a time, hydrogen was replaced, stirring at 15psi and 20℃for 12hr. LCMS showed complete reaction, the reaction solution was filtered, the filtrate was concentrated to dryness, and the crude product was isolated and purified by column chromatography (dichloromethane: methanol=5:1) to give 7- (4-fluorophenyl) heptan-1-amine (155-2) (100 mg,0.454mmol, 46.58%) as a colorless oil .1H NMR(500MHz,CDCl3)δ7.14-7.06(m,2H),6.99-6.91(m,2H),2.68(t,J＝7.1Hz,2H),2.60-2.53(m,2H),1.60-1.55(m,4H),1.43(q,J＝6.9Hz,2H),1.34-1.28(m,6H).LC-MS[M+H]+＝210.1.
Third step referring to the third step in example 154, 7- (3-fluorophenyl) hept-1-amine was changed to 7- (4-fluorophenyl) hept-1-amine to give N- [7- (3-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (155).1H NMR(400MHz,CD3OD)δ7.16(dd,J＝8.5,5.5Hz,2H),6.96(t,J＝8.8Hz,2H),3.20-3.05(m,3H),2.59(t,J＝7.6Hz,2H),2.52-2.42(m,1H),2.34-2.17(m,3H),2.16-2.08(m,1H),2.06-1.95(m,2H),1.89(d,J＝12.3Hz,1H),1.82-1.67(m,3H),1.67-1.58(m,7H),1.51-1.41(m,3H),1.39-1.27(m,7H),1.10-1.01(m,2H),0.98-0.94(m,3H),0.90-0.86(m,1H),0.85(s,3H),0.83(br s,1H),0.80-0.77(m,3H).LC-MS[M+H]+＝526.4.
Example 156
Example 156 preparation of N- [7- (2-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
Reference example 155A conversion of p-fluorobromobenzene to o-fluorobromobenzene in the first reaction step and the same procedure was followed in three steps to give N- [7- (2-fluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide as a compound (156).1H NMR(400MHz,CDCl3)δ7.20-7.13(m,2H),7.05(t,J＝7.2Hz,1H),7.00(t,J＝9.1Hz,1H),5.44(br s,1H),3.27-3.19(m,3H),2.63(t,J＝7.6Hz,2H),2.48-2.40(m,1H),2.38-2.21(m,2H),2.19-2.09(m,2H),2.06-1.85(m,4H),1.82-1.71(m,2H),1.70-1.67(m,2H),1.64-1.55(m,6H),1.48(td,J＝7.1,13.8Hz,2H),1.35-1.24(m,5H),1.10-0.95(m,5H),0.94-0.77(m,9H).LC-MS[M+H]+＝526.4.
Example 157
Example 157 preparation of N- [7- (2, 5-difluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide
Reference example 155 conversion of p-fluorobromobenzene to 2, 5-difluorobromobenzene in the first reaction step and the same procedure was followed in three steps to give N- [7- (2, 5-difluorophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (157).1H NMR(400MHz,CDCl3)δ6.98-6.81(m,3H),5.49(br s,1H),3.28-3.20(m,3H),2.61(t,J＝7.7Hz,2H),2.51-2.22(m,3H),2.19-2.08(m,2H),2.07-2.00(m,1H),1.97-1.88(m,6H),1.80-1.60(m,9H),1.53-1.45(m,2H),1.44-1.37(m,1H),1.33--1.26(m,2H),1.10-0.96(m,5H),0.95-0.79(m,9H).LC-MS[M+H]+＝544.4.
Example 158
Preparation of Compound 158 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (10,10,10-trifluorodecyl) propionamide
In the first step, 9-bromo-1-nonene (1.11 g,5.399 mmol) was added to a reaction flask containing dimethyl sulfoxide (10 mL), and after adding phthalimide potassium salt (1 g,5.399mmol,1 eq), the mixture was stirred at room temperature (20 ℃ C.) for 12hrs; TLC (petroleum ether: ethyl acetate=1:1) showed dot formation with increased polarity; the reaction solution was added with water (30 mL), extracted with dichloromethane (20 mL x 5), the organic phases were combined, washed with water (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a white solid; the crude product was purified by column chromatography (petroleum ether: ethyl acetate=10:1) to give the product 2- (non-8-enyl) isoindole-1, 3-dione (158-1) (1.4 g,4.901mmol, 90%) as a white solid .1H NMR(400MHz,CDCl3)δ7.80-7.72(m,2H),7.68-7.59(m,2H),5.75–5.69(m,1H),4.96-4.80(m,2H),3.60(t,J＝7.2Hz,2H),1.99-1.89(m,2H),1.67-1.51(m,2H),1.37-1.19(m,8H).
The second step was operated in a glove box as follows: to the reaction flask was added 2- (non-8-enyl) isoindole-1, 3-dione (100 mg,0.369 mmol), bis [2- (2, 4-difluorophenyl) -5-trifluoromethylpyridine ] [2-2' -bis (4-t-butylpyridine) ] iridium bis (hexafluorophosphoric acid) salt (8.3 mg,0.007 mmol), sodium triflate (115 mg,0.737 mmol), followed by methanol (5 mL) and stirring in the dark for 3min. The solution was irradiated with blue light (36W, 440 nm) for 12h under 30℃stirring. The reaction solution was concentrated to dryness under reduced pressure, and the crude product was purified by preparative plate separation (petroleum ether: ethyl acetate=5:1) to give 2- (10,10,10-trifluorodecyl) isoindole-1, 3-dione (158-2) (90 mg,0.250mmol, 68%) as a white solid .1H NMR(400MHz,CDCl3)δ7.94-7.82(m,2H),7.79-7.67(m,2H),3.70(t,J＝7.2Hz,2H),2.14-2.00(m,2H),1.69–1.67(m,2H),1.58-1.46(m,2H),1.43-1.21(m,10H).
In a third step, the compound 2- (10,10,10-trifluorodecyl) isoindole-1, 3-dione (158-2) (100 mg,0.293mmol,1.0 eq) was dissolved in ethanol (5 mL), then hydrazine hydrate (0.014 mL,0.293 mmol) was added thereto, the resulting solution was stirred at 50℃for 2h, LC-MS detection was performed, after completion of the reaction the solid was filtered off, and the filtrate was dried by spin-drying to give 10,10,10-trifluorodec-1-amine (158-3) (50 mg,0.154mmol, 52%) as an oily compound .1H NMR(400MHz,CDCl3)δ2.90(br s,2H),2.14-1.98(m,2H),1.72-1.47(m,4H),1.41-1.18(m,10H).LC-MS[M+H]+＝212.1.
Fourth step referring to the third step in example 154, 7- (3-fluorophenyl) hept-1-amine was changed to 10,10,10-trifluorodec-1-amine to give 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (10,10,10-trifluorodecyl) propanamide (158).1H NMR(400MHz,MeOD)δ3.24-3.04(m,3H),2.53-2.41(m,1H),2.36-2.18(m,3H),2.17-2.07(m,3H),2.07-1.95(m,2H),1.90(d,J＝13.6Hz,1H),1.82-1.65(m,4H),1.63(s,4H),1.58-1.44(m,5H),1.44-1.28(m,11H),1.15-1.00(m,2H),0.99-0.94(m.3H),0.91-0.83(m.5H),0.81-0.76(m.3H).LC-MS[M+H]+＝528.4.
Example 159
Preparation of Compound 159N- (1-ethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
Reference example 146 substitution of [ (2-chloro-2, 2-difluoroacetyl) oxy ] sodium in the first step with ethyl iodide, and the same four-step reaction was conducted to give N- (1-ethylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide (159).1H NMR(400MHz,CD3OD)δ7.77(d,J＝1.7Hz,1H),7.35(d,J＝8.8Hz,1H),7.27-7.20(m,2H),6.40(d,J＝3.1Hz,1H),4.21(q,J＝7.2Hz,2H),3.17(dd,J＝10.7,5.5Hz,1H),2.42-2.33(m,2H),2.29-2.16(m,3H),2.12-1.98(m,3H),1.97-1.77(m,4H),1.77-1.66(m,5H),1.65(s,3H),1.64-1.61(m,1H),1.44(t,J＝7.2Hz,3H),1.16-1.04(m,2H),0.97(s,3H),0.93(br d,J＝4.4Hz,1H),0.88(s,3H),0.80(s,3H).LC-MS[M+H]+＝491.4.
Example 160
Preparation of Compound 160 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (undec-2-yl) propanamide
In the first step 2-undecanone (500 mg,2.936 mmol), ammonium acetate (2263.07 mg,29.360 mmol) and sodium cyanoborohydride (129.15 mg,2.055 mmol) were added sequentially to methanol (5 mL), the reaction was stirred at 15℃for 18hr, the pH was adjusted with concentrated hydrochloric acid <2, the solvent was spun dry to give crude product, water (15 mL) was added, extracted with diethyl ether (20 mL), the pH of the aqueous phase was adjusted with potassium hydroxide solid >12, and extracted with diethyl ether (20 mL. Times.3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated to give an oil, which was added with powdered calcium hydride, stirred for 2h, and distilled under reduced pressure to give 2-undecylamine (160-1) (240 mg,1.192mmol, 40.56%) as a white solid .1H NMR(400MHz,DMSO-d6)δ8.07(br s,2H),3.19-2.98(m,1H),1.65-1.53(m,1H),1.47-1.35(m,1H),1.24(br s,13H),1.19-1.13(m,3H),0.90-0.81(m,3H).
Secondly, adding the compound 3- [ (4 aS,4 bR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (39.14 mg,0.117 mmol) into a reaction bottle containing N, N-dimethylformamide (3 mL), adding HATU (88.77 mg,0.233 mmol) and N, N-diisopropylethylamine (0.077 mL,0.443 mmol) and stirring at room temperature (20 ℃) for 0.5hr, adding 2-undecylamine (160-1) (2.69 mg,0.0157 mmol) and stirring for 18hr; the reaction liquid is monitored by LC-MS to have no raw material residue; the reaction mixture was added with water (15 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (20 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was dried under reduced pressure (water pump, 40 ℃), to give a colorless oil; the crude product was chromatographed to give the product 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (undec-2-yl) propanamide (160) (12.94 mg,0.027mmol, 22.73%) as a white solid .1H NMR(500MHz,CD3OD)δ＝3.89-3.77(m,1H),3.16(dd,J＝10.8,5.3Hz,1H),2.55-2.43(m,1H),2.34 -2.17(m,3H),2.17-2.09(m,1H),2.09-1.96(m,2H),1.95-1.86(m,1H),1.83-1.67(m,4H),1.66-1.57(m,5H),1.47 -1.38(m,3H),1.35-1.25(m,16H),1.13-1.06(m,4H),0.98-0.94(m,3H),0.90(t,J＝6.9Hz,4H),0.86(s,3H),0.85 -0.83(m,1H),0.82-0.78(m,3H).LC-MS(ESI)[M+H]+＝488.4.
Example 161
Example 161 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (9-methyldecyl) propionamide
In the first step 9-methyl-n-decanol (100 mg,0.580 mmol) and triphenylphosphine (228.19 mg,0.970 mmol) were added
Phthalimide (93.9 mg, 0.428 mmol) was added sequentially to THF (5 mL), cooled to 0deg.C, nitrogen blanketed, and diisopropyl azodicarboxylate (234.56 mg,1.160 mmol) was then added slowly dropwise. The reaction solution was stirred at 20℃for 12hr.
The reaction was dried by spin and the crude product was purified by hanging plate to give 2- (9-methyldecyl) isoindole-1, 3-dione (161-1) (80 mg,0.266mmol, 41.16%) as a white solid.
In a second step, the compound 2- (9-methyldecyl) isoindole-1, 3-dione (161-1) (80 mg,0.266 mmol) was dissolved in ethanol (3 mL), then hydrazine hydrate (0.1 mL,0.796 mmol) was added thereto, the resulting solution was stirred at 80℃for 3h, the reaction mixture was quenched with hydrochloric acid (1N, 3 mL), then extracted with ethyl acetate (10 mL. Times.2), sodium hydroxide (15 wt%) was added to the aqueous phase to adjust the pH to 8, then extracted with dichloromethane (10 mL. Times.4), the organic phases combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give 9-methyldec-1-amine (161-2) (18 mg,0.105mmol, 39.59%) as a colorless oil, which was used directly in the next step.
Third step referring to the third step in example 154, 7- (3-fluorophenyl) hept-1-amine was changed to 9-methyldec-1-amine to give 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (9-methyldecyl) propanamide (161).1H NMR(400MHz,MeOD)δ8.46(d,J＝5.7Hz,1H),7.86(t,J＝8.2Hz,1H),7.41(d,J＝7.5Hz,1H),7.34(s,1H),7.19(d,J＝8.7Hz,1H),7.06(t,J＝7.8Hz,1H),6.96(d,J＝6.7Hz,1H),3.64(s,1H),2.89-2.72(m,8H),2.42(d,J＝7.9Hz,2H),2.02-2.01(m,2H),1.70(d,J＝5.7Hz,7H),1.61(d,J＝7.0Hz,10H),1.17(s,3H).LC-MS[M+H]+＝488.5.
Example 162
EXAMPLE 162 preparation of 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ 1-methyl-3- (methyldioxy-lambda 6 -thio) indol-5-yl ] butanamide
In the first step, dimethyl disulfide (2.6 g,27.6 mmol), phthalimide (7.35 g,50 mmol) was added sequentially to a mixture of hot pyridine (20 mL) and acetonitrile (25 mL), stirred for 10 minutes, then placed under an ice bath, and elemental bromine (5 g,1.72mL,31.3 mL) was slowly added dropwise, followed by stirring at room temperature for 1hr, cooled to 0℃and quenched by dropwise with water (100 mL). The mixture was stirred for 1hr, a large amount of solid was precipitated, and 2- (methylthio) isoindole-1, 3-dione (162-1) (7.95 g,40.322mmol, 80.72%) was obtained as a red solid by filtration. 1H NMR(CD3 OD,400 MHz) delta 7.8-8.0 (m, 4H), 2.50 (s, 3H).
In the second step, 5-bromo-1-methylindole (489.30 mg,2.329 mmol), 2- (methylsulfanyl) isoindole-1, 3-dione (162-1) (500 mg,2.588 mmol) and magnesium bromide (23.82 mg,0.129 mmol) were added sequentially to N, N-dimethylformamide (6 mL), and after nitrogen substitution, stirred at 90℃for 16hr. TLC, indicating completion of the reaction. The temperature was reduced to 0 ℃, quenched with sodium hydroxide (1 n,4 ml) and extracted with ethyl acetate (5 ml x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=5:1) to give 5-bromo-1-methyl-3- (methylsulfanyl) indole (162-2) (353 mg,1.240mmol, 47.92%) as a yellow oil .1H NMR(CDCl3,400MHz)δ7.87(d,J＝1.8Hz,1H),7.34(dd,J＝8.6,1.9Hz,1H),7.19(d,J＝8.6Hz,1H),7.17(s,1H),3.76(s,3H),2.34(s,3H).
In a third step 5-bromo-1-methyl-3- (methylsulfanyl) indole (162-2) (170 mg, 0.264 mmol) was dissolved in tetrahydrofuran (3 mL), a solution of potassium peroxymonosulphonate (2040 mg,3.318 mmol) in water (4.5 mL) was added, stirred at 25℃for 16hr, after completion of the reaction, diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give the crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=3:1) to give 5-bromo-1-methyl-3- (methyldioxy-lambda 6 -thio) indole (162-3) (100 mg,0.312mmol, 47.06%) as a yellow solid .1H NMR CDCl3,400MHz)δ8.10(d,J＝1.7Hz,1H),7.69(s,1H),7.47(dd,J＝8.8,1.8Hz,1H),7.28(d,J＝8.8Hz,1H),3.86(s,3H),3.15(s,3H).
Fourth step in reference example 102, in the second step, 5-bromo-4-fluoro-1-methylindole was changed to 5-bromo-1-methyl-3- (methyldioxy-lambda 6 -thio) indole to give 5- [ (diphenylmethylidene) amino ] -1-methyl-3- (methyldioxy-lambda 6 -thio) indole (162-4).1H NMR(CDCl3,400MHz)δ7.78(d,J＝7.2Hz,2H),7.58(s,1H),7.50(d,J＝6.9Hz,1H),7.46-7.41(m,2H),7.28-7.23(m,4H),7.21-7.15(m,3H),6.93(dd,J＝8.8,1.9Hz,1H),3.81(s,3H),2.92(s,3H).
Fifth step referring to the third step in example 102, 5- [ (diphenylmethylene) amino ] -4-fluoro-1-methylindole was changed to 5- [ (diphenylmethylene) amino ] -1-methyl-3- (methyldioxy-lambda 6 -thio) indole to give 1-methyl-3- (methyldioxy-lambda 6 -thio) indol-5-amine (162-5). LC-MS [ m+h ] + =225.0.
In a sixth step 4- [ (4 bR, 7S) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydro phenanthryl ] butanoic acid (II) (10 mg,0.022 mmol), 1-methyl-3- (methyldioxy-lambda 6 -thio) indol-5-amine (162-5) (9.7 mg,0.043 mmol) was added to a reaction flask containing anhydrous acetonitrile (1 mL), N-methylimidazole (0.01 mL,0.076 mmol) was added thereto, the mixture was stirred at 25℃for 0.5hr, N, N ', N' -tetramethylchlorofluorophosphate (9.9 mg,0.032 mmol) was then stirred at 25℃for 2hr, S detection reaction was completed, water was added thereto, and then the crude product was extracted with dichloromethane (10 mL), dried by filtration, the product 4- [ (4 ar,4bs,7r,8as,10 as) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ 1-methyl-3- (methyldioxy- λ6-thio) indol-5-yl ] butanamide (162-6) (13 mg,0.017mmol, 76.43%) was then isolated and purified by preparative plate (petroleum ether: ethyl acetate=1:1) as a white solid.
In the seventh step, 4- [ (4 aR,4bS,7R,8aS,10 aS) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ 1-methyl-3- (methyldioxy-. Lamda.6-thio) indol-5-yl ] butanamide (162-6) (13 mg,0.017 mmol) was added to methanol (1 mL), and camphorsulfonic acid (6.9 mg,0.030 mmol) was added thereto and the reaction mixture was stirred at 25℃for 16hr. The reaction solution was purified by preparative chromatography to give 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- [ 1-methyl-3- (methyldioxy-lambda 6 -thio) indol-5-yl ] butanamide (162) (3.45 mg,0.006mmol, 37.66%) as a white solid .1H NMR(CD3OD,400MHz)δ8.04-7.93(m,1H),7.82-7.72(m,1H),7.47-7.36(m,2H),3.85-3.71(m,3H),3.13-2.99(m,4H),2.33-2.22(m,2H),2.19-2.01(m,3H),1.95-1.57(m,8H),1.55-1.51(m,4H),1.36-1.18(m,3H),1.06-0.90(m,2H),0.88-0.83(m,3H),0.79-0.71(m,5H),0.71-0.65(m,3H).LC-MS[M+H]+＝555.3.
Example 163
Example 163 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (2, 5, 8-trioxadec-10-yl) propanamide
Reference example 36 conversion of 1-methylindole-5-amine to 2- (2, 5-dioxahept-7-yloxy) ethan-1-amine gave the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (2, 5, 8-trioxadec-10-yl) propanamide (163).1H NMR(400MHz,MeOD)δ3.66-3.58(m,6H),3.57-3.51(m,4H),3.36(s,3H),3.34(d,J＝5.6Hz,2H),3.16(dd,J＝10.4,5.6Hz,1H),2.53-2.40(m,1H),2.30 2.19(m,2H),2.18-2.09(m,1H),2.07-1.85(m,4H),1.83-1.67(m,4H),1.65-1.60(m,4H),1.47-1.31(m,2H),1.13-1.01(m,3H),0.99-0.94(m,3H),0.86(s,4H),0.82-0.78(m,3H).LC-MS[M+H]+＝480.4.
Example 164
Example 164 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (2-methylphenyl) heptyl ] propanamide
First step referring to the first step in example 155, p-bromofluorobenzene was changed to o-iodotoluene (500 mg,2.293 mmol) to give 7- (2-methylphenyl) heptanenitrile (164-1) (200 mg,0.894mmol, 38.99%) as a colorless oil .1H NMR(400MHz,CD3OD)δ7.24-6.89(m,4H),2.69-2.56(m,2H),2.47-2.38(m,2H),2.29(s,3H),1.69-1.54(m,4H),1.52-1.39(m,4H).
The second step compound 7- (2-methylphenyl) heptanenitrile (164-1) (200 mg,0.894mmol, 38.99%) was used for flow chemistry reactor hydrogenation, parameters 14% Ni/Al 2O3, 15V MeOH,120 ℃,2.5MPa.TLC indicated starting material exhaustion, new spot formation. The reaction mixture was concentrated to give 7- (2-methylphenyl) hept-1-amine (164-2) (158 mg,0.769mmol, 77.45%) as a colorless oil .1H NMR(400MHz,CD3OD)δ7.18-6.96(m,4H),2.70-2.53(m,4H),2.34-2.20(m,3H),1.62-1.45(m,4H),1.37(s,6H).
Third step referring to the third step in example 154, 7- (3-fluorophenyl) hept-1-amine was changed to 7- (2-methylphenyl) hept-1-amine to give 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (2-methylphenyl) heptyl ] propionamide (164).1H NMR(400MHz,CD3OD)δ7.18-6.95(m,4H),3.21-3.10(m,2H),2.66-2.55(m,2H),2.52-2.40(m,1H),2.28(s,3H),2.27-2.18(m,2H),2.16-2.07(m,1H),2.06-1.97(m,2H),1.89(d,J＝12.4Hz,2H),1.79-1.67(m,3H),1.62(s,3H),1.57(d,J＝7.5Hz,2H),1.51-1.44(m,2H),1.43-1.26(m,10H),1.11-1.00(m,2H),0.98-0.93(m,3H),0.89(d,J＝6.6Hz,2H),0.87-0.82(m,4H),0.82-0.75(m,3H).LC-MS[M+H]+＝522.4.
Example 165
Example 165 preparation of 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (7- {2- [ (trifluoromethyl) oxy ] phenyl } heptyl) propanamide
Reference example 164 conversion of o-iodotoluene to o-trifluoromethoxybenzene gives the compound 3- [ (4 aS,4bR,7S,8aR, 10R) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (7- {2- [ (trifluoromethyl) oxy ] phenyl } heptyl) propanamide (165).1H NMR(400MHz,CD3OD)δ7.40-7.29(m,1H),7.29-7.21(m,3H),3.21-3.12(m,2H),2.73-2.63(m,2H),2.53-2.41(m,1H),2.26-2.18(m,2H),2.16-2.08(m,1H),2.05-1.98(m,2H),1.94-1.83(m,2H),1.80-1.68(m,3H),1.67(s,3H),1.54(d,J＝2.8Hz,1H),1.52-1.43(m,3H),1.39-1.28(m,9H),1.06(dt,J＝8.0,4.4Hz,2H),0.98-0.93(m,3H),0.89(d,J＝6.0Hz,2H),0.87-0.84(m,4H),0.82-0.77(m,3H).LC-MS[M+H]+＝592.4.
Example 166
Example 166 preparation of 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (1-phenylindol-5-yl) butanamide
Reference example 40 conversion of 1-methylindole-5-amine to 1-phenylindol-5-amine gives 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthryl ] -N- (1-phenylindol-5-yl) butanamide (166).1H NMR(400MHz,CD3OD)δ7.89(d,J＝1.2Hz,1H),7.58-7.51(m,4H),7.49-7.43(m,2H),7.37(t,J＝6.4Hz,1H),7.27(dd,J＝8.8,1.6Hz,1H),6.63(d,J＝3.2Hz,1H),3.15(dd,J＝10.4,5.6Hz,1H),2.37(dt,J＝7.2,2.4Hz,2H),2.26-2.14(m,3H),2.06-1.91(m,3H),1.63(s,10H),1.49-1.27(m,4H),1.13-1.02(m,2H),0.95(s,3H),0.85(s,3H),0.78(s,3H).LC-MS[M+H]+＝539.4.
Example 167
Example 167 preparation of 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3-hydroxy-1-methylindol-5-yl) butanamide
The compound N- (3-bromo-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (171) (20 mg,0.030 mmol) was dissolved in methanol (3 mL), and camphorsulfonic acid (12.5 mg,0.054 mmol) was added and reacted at 25℃for 12 hours. The reaction solution was cooled to room temperature and then dried by spin to give a crude product, which was then separated by preparative chromatography to give 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (3-hydroxy-1-methylindol-5-yl) butanamide (167) (4.69 mg, 0.399 mmol, 31.37%) as a colorless oil .1H NMR(400MHz,CD3OD)δ7.56(s,1H),7.44(dd,J＝8.4,2.0Hz,1H),6.92(d,J＝8.4Hz,1H),3.20(s,3H),3.15(dd,J＝10.8,5.6Hz,1H),2.37-2.28(m,2H),2.23-2.13(m,2H),2.06-1.95(m,2H),1.94-1.86(m,1H),1.78(d,J＝12.8Hz,2H),1.72-1.63(m,4H),1.61(s,4H),1.49-1.27(m,3H),1.10-0.99(m,2H),0.97-0.94(m,3H),0.89(d,J＝6.8Hz,1H),0.87-0.82(m,5H),0.80-0.77(m,3H).LC-MS[M+H]+＝499.3.
Example 168
Example 168 preparation of N- (1-Acetylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide
Reference example 40 conversion of 1-methylindole-5-amine to 1-acetylindol-5-amine gives N- (1-acetylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (168).1H NMR(400MHz,CD3OD)δ8.30(d,J＝8.8Hz,1H),7.92(d,J＝2.0Hz,1H),7.68(d,J＝3.6Hz,1H),7.35(dd,J＝8.8,2.0Hz,1H),6.65(d,J＝3.6Hz,1H),3.15(dd,J＝10.8,5.6Hz,1H),2.64(s,3H),2.40-2.35(m,2H),2.26-2.13(m,3H),2.05-1.96(m,2H),1.93-1.87(m,1H),1.84-1.75(m,2H),1.73-1.60(m,9H),1.48-1.40(m,1H),1.13-1.02(m,2H),0.95(s,3H),0.91-0.83(m,6H),0.78(s,3H).LC-MS[M+H]+＝505.4.
Example 169
Example 169 preparation of N- (3-fluoro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanamide
In the first step, the compound 3, 3-difluoro-5-nitro-2, 3-dihydro-1H-indol-2-one (1 g, 4.640 mmol) was dissolved in tetrahydrofuran (15 mL) and borane tetrahydrofuran (23.350 mL,23.350 mmol) was added under nitrogen at 0deg.C. Then the temperature was raised to 70℃and stirred for 5 hours. TLC (petroleum ether/ethyl acetate=3/1) showed the reaction was complete and a new spot was formed. The reaction was quenched with saturated sodium bicarbonate solution (10 mL), extracted with ethyl acetate (20 mL x 3), the organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and dried by spinning. Purification of the crude product by silica gel column chromatography (PE: ea=5:1) gave 3-fluoro-5-nitro-1H-indole (169-1) (1 g,2.220mmol, 47.55%) as a yellow solid .1H NMR(400MHz,CDCl3)δ8.38(s,1H),8.25(dd,J＝9.0,2.0Hz,1H),8.16-8.09(m,1H),7.17(t,J＝2.8Hz,1H).
In a second step, the compound 3-fluoro-5-nitro-1H-indole (169-1) (800 mg,4.441 mmol) was dissolved in N, N-dimethylformamide (10 mL), and sodium hydrogen (355.28 mg,8.882 mmol) was added at 0deg.C under nitrogen and stirred for 10 min. Methyl iodide (0.829 mL,13.323 mmol) was then added and stirred at 0deg.C for 1 hour under nitrogen. LCMS monitored the formation of product, spotting the end of the reaction and a new spot. The reaction was quenched by dropwise addition of dilute hydrochloric acid (1M) in an ice bath, ph=7 was adjusted, extracted with ethyl acetate (20 ml×3), and the organic phase was washed with saturated brine (20 ml×3), dried over anhydrous sodium sulfate, filtered, and dried by spinning. The crude product was purified by preparative chromatography to give 3-fluoro-1-methyl-5-nitroindole (169-2) (740 mg, 3.81mmol, 85.82%) as a yellow solid .1H NMR(400MHz,CDCl3)δ8.62(d,J＝2.0Hz,1H),8.15(dd,J＝9.2,2.0Hz,1H),7.32(dd,J＝9.2,2.0Hz,1H),7.01(d,J＝2.8Hz,1H),3.81(s,3H).19F NMR(376MHz,CDCl3)δ-171.63(s,1F).LC-MS[M+H]+＝195.0.
In a third step, the compound 3-fluoro-1-methyl-5-nitroindole (169-2) (740.00 mg, 3.81mmol) was dissolved in water (10 mL), methanol (10 mL) and tetrahydrofuran (10 mL), and zinc powder (3220 mg,49.251 mmol) and ammonium chloride (2040.00 mg,38.111 mmol) were added at 0deg.C. The reaction was stirred at 0℃for 1 hour. LCMS showed the reaction was complete and product formation was detected. The reaction was filtered, diluted hydrochloric acid (1 mL) was added to the filtrate, and the mixture was dried by spinning. Purification of the crude product by preparative chromatography gave 3-fluoro-1-methylindol-5-amine (169-3) (440 mg,2.680mmol, 70.32%) as a grey solid. LC-MS [ m+h ] + =164.9.
In a fourth step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (10 mg,0.022 mmol) was dissolved in acetonitrile (1 mL) and N, N, N, N-tetramethyl chloroformidine hexafluorophosphate (7.28 mg,0.026 mmol) and N-methylimidazole (6.21 mg,0.076 mmol) were added at 25℃and stirred for 10 minutes at 25 ℃. 3-fluoro-1-methylindol-5-amine (169-3) (7.10 mg,0.043 mmol) was then added to the reaction system, and the reaction was stirred at 25℃for 16 hours. LCMS showed the reaction was complete and product formation was detected. TLC (petroleum ether: ethyl acetate=5:1) showed new spot formation. The reaction solution was spin-dried and purified by TLC plate (petroleum ether: ethyl acetate=5:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (3-fluoro-1H-indol-5-yl) butanamide (169-4) (10 mg,0.015mmol, 68.75%) as a white solid .1H NMR(400MHz,CDCl3)δ7.75(d,J＝1.6Hz,1H),7.37-7.30(m,1H),7.21(dd,J＝9.2,2.0Hz,1H),6.84(d,J＝2.8Hz,1H),3.71(s,3H),3.20(d,J＝4.4Hz,1H),2.40-2.29(m,4H),2.19-2.08(m,3H),1.94-1.85(m,5H),1.74-1.67(m,7H),1.63(s,3H),1.53(dd,J＝13.2,4.0Hz,3H),1.26(s,1H),0.88(s,3H),0.85-0.79(m,9H),0.78-0.74(m,6H),0.04(s,6H).LC-MS[M+H]+＝609.5.
In a fifth step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (3-fluoro-1-methylindol-5-yl) butanamide (169-4) (10 mg,0.016 mmol) was dissolved in methanol (2 mL) and (1S) - (+) -10-camphorsulfonic acid (7.63 mg,0.033 mmol) was added at 25 ℃. The reaction was stirred at 25℃for 16 hours. LCMS showed the reaction was complete and product formation was detected. The reaction solution was dried by spinning, and the crude product was purified by preparative chromatography to give N- (3-fluoro-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (169) (1.05 mg,0.002mmol, 12.93%) as a white solid .1H NMR(400MHz,CD3OD)δ7.79(s,1H),7.35-7.19(m,2H),7.02(d,J＝2.4Hz,1H),3.72(s,3H),3.18-3.12(m,1H),2.43-2.30(m,2H),2.26-2.08(m,4H),2.04(s,2H),1.94(s,1H),1.78(d,J＝13.2Hz,2H),1.69(d,J＝13.6Hz,3H),1.63(s,6H),1.29(s,2H),0.95(s,3H),0.86(s,4H),0.84(s,1H),0.78(s,3H).19F NMR(376MHz,CD3OD)δ-178.88(s,1F).LC-MS[M+H]+＝495.4.
Example 170
EXAMPLE 170 preparation of 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (imidazo [1,2-a ] pyridin-6-yl) butanamide
Reference example 40 conversion of 1-methylindole-5-amine to imidazo [3,2-a ] pyridin-6-amine gives 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- (imidazo [1,2-a ] pyridin-6-yl) butanamide (170).1H NMR(400MHz,CD3OD)δ8.30(d,J＝8.8Hz,1H),7.92(d,J＝2.0Hz,1H),7.68(d,J＝3.6Hz,1H),7.35(dd,J＝8.8,2.0Hz,1H),6.65(d,J＝3.6Hz,1H),3.15(dd,J＝10.8,5.6Hz,1H),2.64(s,3H),2.40-2.35(m,2H),2.26-2.13(m,3H),2.05-1.96(m,2H),1.93-1.87(m,1H),1.84-1.75(m,2H),1.73-1.60(m,9H),1.48-1.40(m,1H),1.13-1.02(m,2H),0.95(s,3H),0.91-0.83(m,6H),0.78(s,3H).LC-MS[M+H]+＝505.4.
Example 171
Example 171 preparation of N- (3-bromo-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] butanamide
In the first step, the compound 3-bromo-1-methyl-5-nitroindole (100 mg, 0.298 mmol) was dissolved in methanol (3 mL), tetrahydrofuran (3 mL) and water (3 mL), and then zinc powder (256.3 mg,3.920 mmol) and ammonium chloride (209.7 mg,3.920 mmol) were added, respectively, at 0deg.C. The nitrogen was replaced and reacted at 25℃for 1 hour. LCMS showed complete reaction of starting material and monitored for product formation. The residue was filtered through a sand core funnel, washed with 10m ethyl acetate, then dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product as a white solid, 3-bromo-1-methylindol-5-amine (171-1) (72 mg,0.288mmol, 73.43%), which was used directly in the next step. LC-MS [ m+h ] + = 224.9,226.9.
In a second step, the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-YL) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (II) (20 mg,0.043 mmol) and 3-bromo-1-methylindol-5-amine (171-1) (9.7 mg,0.043 mmol) were dissolved in N, N-dimethylformamide (3 mL), to which were then added N-methylimidazole (0.021 mL,0.130 mmol) and O- (7-azabenzotriazol-1-YL) -N, N, N, N-tetramethylurea hexafluorophosphine salt (19.7 mg,0.052 mmol). The reaction was reacted at 25℃for 2h. LCMS showed complete reaction of starting material and monitored for product formation. The reaction solution was diluted with 10mL of water, extracted three times with ethyl acetate (10 mL), washed 2 times with saturated brine (20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was isolated by TCL plate (petroleum ether: ethyl acetate=3:1) to give N- (3-bromo-1-methylindol-5-yl) -4- [ (4 as,4br,7s,8ar,10 ar) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (171-2) (20 mg,0.027mmol, 62.18%) as a colorless oil. LC-MS [ m+h ] + = 671.3.
In a third step, the reactant N- (3-bromo-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7- { [ dimethyl (2-methylpropan-2-yl) silyl ] oxy } -2,4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (171-2) (15 mg,0.022 mmol) was dissolved in tetrahydrofuran (2 mL), and a solution of tetrabutylammonium fluoride in tetrahydrofuran (0.3 mL,0.3mmol,1 mol/L) was added. The reaction system was replaced with a nitrogen atmosphere. Stirred at 70℃for 12 hours. The reaction solution was cooled to room temperature and concentrated to give a crude product, and N- (3-bromo-1-methylindol-5-yl) -4- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanamide (171) (4.82 mg,0.009mmol, 38.36%) was obtained as a colorless oil by separation by preparative chromatography .1H NMR(400MHz,CD3OD)δ7.75(d,J＝4.0Hz,1H),7.38-7.34(m,1H),7.25(s,1H),3.79(s,3H),3.20-3.11(m,1H),2.42-2.33(m,2H),2.32-2.08(m,3H),2.08-1.94(m,2H),1.92(s,3H),1.73-1.60(m,8H),1.43(d,J＝10.0Hz,1H),1.35-1.27(m,2H),1.17-0.99(m,2H),0.95(s,3H),0.89-0.83(m,5H),0.80-0.75(m,3H).LC-MS[M+H]+＝555.3.
Example 172
Example 172 preparation of N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazapentan-5-yl) amino ] methanamide
In the first step, after potassium tert-butoxide (12.6 g,112 mmol) was dissolved in tert-butanol (100 mL), 17- (2-oxypropylene) androsta-5 (4) -en-3-one (5 g,15.1 mmol) was added as starting material at 15℃in a water bath. After the addition was completed stirring was continued for 15 minutes, methyl iodide (9.42 mL,151 mmol) was added dropwise followed by stirring at 25℃for 18 hours. The reaction was added to ice water (50 mL), the organic phase was concentrated together with the aqueous phase until a large amount of white solid appeared, filtered, the filter cake was dissolved with dichloromethane/methanol (1:1, 300 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give (3 as,3br,9ar,9bs,11 as) -1-hydroxy-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthren-7-one (172-1) (1.8 g,5.12mmol, 33.83%) as a white solid .1H NMR(400MHz,CDCl3)δ5.55(dd,J＝5.2,2.4Hz,1H),3.66(dt,J＝8.4,6.0Hz,1H),2.62-2.41(m,2H),2.15-2.02(m,3H),1.89-1.83(m,1H),1.86(td,J＝12.0,3.6Hz,1H),1.68-1.62(m,2H),1.54-1.28(m,3H),1.23(d,J＝1.6Hz,6H),1.19-0.91(m,4H),0.87(s,3H),0.77(s,3H).
In the second step (3 aS,3bR,9aR,9bS,11 aS) -1-hydroxy-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10, -11,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthren-7-one (172-1) (3.1 g,9.795 mmol) was dissolved in anhydrous methanol (55 mL) and dichloroethane (55 mL) followed by addition of ammonium acetate (7.55 g,97.9 mmol) and sodium cyanoborohydride (1.23 g,19.6 mmol). The reaction was stirred at 40 ℃ for 18 hours, thin layer chromatography plate (petroleum ether/ethyl acetate=3:1) indicated that there were multiple new spots left to form. The organic phase was concentrated to a white solid which was added water (40 mL). After the aqueous phase was adjusted to pH 9 with 1 molar aqueous sodium hydroxide, the solution was filtered, the filter cake was dissolved with dichloromethane/methanol (1:1, 50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude (3 aS,3bR,7S,9aR,9bS,11 aS) -7-amino-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthren-1-ol (172-2) (3.72 g,4.967mmol, 50.71%) as a white solid, and the crude purity of LC-MS [ M+H ] + =318.5.
In the third step, triethylamine (2.82 mL,20.3 mmol) and (3 aS,3bR,7S,9aR,9bS,11 aS) -7-amino-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthren-1-ol (172-2) (3.22 g,10.1 mmol) were dissolved in a mixed solution of dichloromethane (18 mL) and methanol (18 mL), and di-t-butyl dicarbonate (2.79 mL,12.2 mmol) was added thereto at 25℃and the reaction was stirred at 25℃for 18 hours. Thin layer chromatography plates (dichloromethane/methanol=10:1) indicated the starting material was exhausted and new spots were formed. After the reaction solution was concentrated, the crude product was separated and purified by column chromatography (PE: ea=4:1) to give { [ (3 as,3br,7s,9ar,9bs,11 as) -1-hydroxy-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthren-7-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-3) (3.23 g,6.19mmol, 61.01%) as a white solid .1H NMR(400MHz,CDCl3)δ5.60-5.44(m,1H),4.44(d,J＝9.6Hz,1H),3.63(t,J＝8.4Hz,1H),3.35-3.25(m,1H),2.16-2.05(m,2H),1.87-1.77(m,1H),1.76-1.60(m,3H),1.59-1.48(m,4H),1.45-1.42(m,9H),1.30-1.19(m,4H),1.12-1.00(m,9H),0.99-0.87(m,2H),0.74(d,J＝1.6Hz,3H).
In a fourth step { [ (3 aS,3bR,7S,9aR,9bS,11 aS) -1-hydroxy-6, 9a,11 a-tetramethyl-2, 3a,3b,4,6,7,8, 9a,9b,10,11 a-decatetrahydro-1H-cyclopenta [1,2-a ] phenanthr-7-yl ] amino } propan-2-yl ester of methane acid (172-3) (4.42 g,10.6 mmol) was dissolved in a mixed solution of anhydrous tetrahydrofuran (26.5 mL) and anhydrous methanol (26.5 mL), and the solution was clarified for fluid chemistry. (fluid chemistry: 5% palladium hydroxide/aluminum oxide, tetrahydrofuran/methanol=1:1.53 mL,0.3mL/min,85 ℃, H2.5 MPa) for 9 hours. Thin layer chromatography plates indicated the depletion of the starting material and the formation of new spots. The organic phase was concentrated to give { [ (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -1-hydroxy-6, 9a,11 a-tetramethylhexadeca-hydro-1H-cyclopenta [1,2-a ] phenanthren-7-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-4) (4.6 g,9.87mmol, 93.21%) as a white solid .1H NMR(400MHz,CD3OD)δ3.55(t,J＝8.8Hz,1H),3.22-3.13(m,1H),2.02-1.91(m,1H),1.85-1.75(m,2H),1.75-1.69(m,1H),1.67-1.57(m,3H),1.54(dd,J＝7.2,3.2Hz,1H),1.51-1.48(m,1H),1.44(s,9H),1.42-1.37(m,1H),1.36-1.18(m,4H),1.16-1.01(m,3H),1.00-0.92(m,2H),0.90(s,3H),0.85(s,3H),0.77(s,3H),0.71(s,3H),0.69-0.60(m,1H).
In the fifth step, starting { [ (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -1-hydroxy-6, 9a,11 a-tetramethylhexadeca-1H-cyclopenta [1,2-a ] phenanthren-7-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-4) (6.53 g,15.6 mmol) was dissolved in methylene chloride (140 mL), and then, dess-martin oxidant (7.92 g,18.7 mmol) was added at 0℃and replaced with nitrogen three times, followed by stirring at 25℃for 1.5 hours. Thin layer chromatography plates (petroleum ether/ethyl acetate=5:1) indicated the starting material was exhausted and new spots were formed. The reaction was added to a saturated Na 2SO3 (40 mL) solution followed by a saturated NaHCO 3 (40 mL) solution and stirred for 5 min, and the reaction was extracted with dichloromethane (40 mL x 3). The organic phase was washed with water (60 ml x 2) and then dried over Na2SO4, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give the product { [ (3 as,3br,5ar,7s,9ar,9bs,11 as) -6, 9a,11 a-tetramethyl-1-oxohexadeca-1H-cyclopenta [1,2-a ] phenanthren-7-yl ] amino } methano-2-methylpropan-2-yl ester (172-5) (4.1 g,8.836mmol, 80.57%) as a white solid .1H NMR(400MHz,CDCl3)δ4.39(d,J＝10.0Hz,1H),3.32-3.21(m,1H),2.42(dd,J＝19.2,8.8Hz,1H),2.11-2.01(m,1H),1.97-1.87(m,2H),1.80-1.75(m,1H),1.71-1.63(m,3H),1.62(s,2H),1.44(s,9H),1.39-1.30(m,2H),1.27-1.20(m,3H),1.10-1.03(m,2H),0.99-0.93(m,2H),0.90(s,3H),0.85(s,3H),0.83(s,3H),0.75(s,3H),0.71-0.64(m,1H).
In a sixth step, the compound { [ (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -6, 9a,11 a-tetramethyl-1-oxolano [1,2-a ] phenanthren-7-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-5) (0.5 g, 1.197mmol) was dissolved in anhydrous dioxane (120 mL), replaced with nitrogen gas by a light reaction apparatus, irradiated with 300W Hg lamp, and reacted at 25℃for 50 minutes. TLC (PE: EA=5:1, phosphomolybdic acid developer) showed that the reaction was complete, the dioxane was directly spin-dried, and the crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 10:1) to give { [ (2S, 4aR,4bS,8aR,10 aR) -8- (2-formylethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthre-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-6) (300 mg,0.64 mmol, 54.00%) as a white solid .1H NMR(400MHz,CDCl3)δ9.81-9.74(m,1H),3.35-3.22(m,1H),2.44(dd,J＝14.4,3.8Hz,2H),2.16-2.08(m,2H),1.92(s,1H),1.71(dd,J＝13.6,3.6Hz,2H),1.61(s,3H),1.44(s,9H),1.33-1.21(m,4H),1.11-1.02(m,4H),0.90(d,J＝3.2Hz,6H),0.88-0.82(m,4H),0.74(s,3H).
In the seventh step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (2-formylethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-6) (300 mg,0.718 mmol) was dissolved in t-butanol (10 mL), and 2-methylbutan-2-ene (0.362 mL,4.310 mmol) under ice-bath and sodium dihydrogen phosphate (258.60 mg,2.155 mmol) and sodium chlorite (227.38 mg,2.514 mmol) were dissolved in 2mL of water to add the reaction system. The ice bath was removed and reacted at 20℃for 2 hours. TLC (PE: ea=5:1, phosphomolybdic acid developer) showed the reaction was complete. The reaction was quenched with saturated sodium sulfite solution (5 mL), extracted with ethyl acetate (5 mL x 3), washed with saturated brine (5 mL x 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to give the crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give 3- [ (4 as,4br,7s,8ar,10 ar) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propionic acid (172-7) (160 mg,0.351mmol, 48.80%) as a white solid LC-MS [ m+h ] + = 578.2.
In an eighth step, the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanoic acid (172-7) (160 mg,0.351 mmol) was dissolved in toluene (3 mL), and triethylamine (0.045 mL,0.323 mmol) and diphenyl azide phosphate (46.65 mg,0.169 mmol) were added at 25℃and stirred at 90℃for 1 hour. 1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-amine (28.70 mg,0.194 mmol) was then added. The reaction was stirred at 25℃for 16 hours. LCMS showed product formation. The reaction was quenched with water (5 mL), extracted with ethyl acetate (5 mL x 3), washed with saturated aqueous sodium chloride (5 mL x 3), and the organic phase was dried over anhydrous sodium sulfate, filtered and dried by spinning. The crude product was isolated by preparative chromatography to give { [ (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- [2- ({ [ (1-methylbenzo [ d ] [1,2,3] triazacyclopenta-5-yl) amino ] carbonyl } amino) ethyl ] -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-8) (35 mg,0.060mmol, 37.46%) as a white solid. LC-MS [ m+h ] + =579.4.
In a ninth step, raw material { [ (2S, 4aS,4bS,8aR,10 aS) -8- (3- { [6- (2, 5-difluorophenyl) hexyl ] amino } -3-oxoypropyl) -4a, 7-dimethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (172-8) (35 mg,0.058 mmol) was dissolved in hexafluoroisopropanol (2 mL). The reaction was carried out at 80℃for 48 hours. LCMS showed complete reaction of starting material and monitored for product formation. The reaction solution was concentrated to obtain a crude product, and the crude product was subjected to preparative chromatography to obtain N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] methanamide (15 mg,0.028mmol, 46.10%) as a white solid. Then separating and purifying by preparative chromatography to obtain N- {2- [ (4 aS,4bR,7S,8aR,10 aR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] ethyl } -1- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopentan-5-yl) amino ] methanamide (172) (3.17 mg,0.006mmol, 20.62%) as white solid .1H NMR(400MHz,CD3OD)δ8.22-8.05(m,1H),7.76(d,J＝8.8Hz,1H),7.51(dd,J＝9.2,2.0Hz,1H),4.37(s,3H),3.30-3.08(m,2H),2.91(dd,J＝10.4,5.6Hz,1H),2.46-2.26(m,2H),2.25-1.93(m,3H),1.93-1.86(m,1H),1.85-1.73(m,3H),1.69(s,3H),1.59(s,1H),1.56-1.41(m,2H),1.41-1.28(m,1H),1.15(dt,J＝12.0,6.4Hz,2H),1.06(s,3H),1.03-0.94(m,2H),0.92(s,2H),0.90(s,3H),0.88(s,1H).LC-MS[M+H]+＝479.4.
Examples 173&175
Example 173 preparation of N- (4-fluorodecyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide
EXAMPLE 175 preparation of N- (4, 4-difluorodecyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
In the first step, the compound 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b, -5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionic acid (III) (100 mg,0.299 mmol) was dissolved in N, N-dimethylformamide (3 mL), followed by addition of N, N, N, N-tetramethyl chloroformidine hexafluorophosphate (109.04 mg,0.389 mmol) and N-methylimidazole (85.91 mg,1.046 mmol) for 0.5 hours at 25 ℃.4, 4-Difluorodec-1-amine (raw material was not pure, containing a part of 4-fluorodec-1-amine) (57.78 mg,0.299 mmol) was added to the reaction, and the reaction was carried out at 25℃for 2 hours. LCMS showed complete reaction of starting material and monitored for product formation. The reaction solution was filtered, and the filtrate was subjected to preparative chromatography to give N- (4, 4-difluorodecyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (175-1) (25 mg,0.029mmol,9.84%, containing fraction 173) as a colorless oil. LC-MS [ m+h ] + = 510.5 and N- (4-fluorodecyl) -3- [ (4 as,4br,7s,8ar,10 ar) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propionamide (173)(10mg,0.020mmol,6.80％).1H NMR(400MHz,CD3OD)δ4.58-4.36(m,1H),3.26-3.07(m,3H),2.54-2.41(m,1H),2.39-2.28(m,1H),2.27-2.19(m,2H),2.18-2.10(m,1H),2.02(d,J＝11.6Hz,2H),1.90(d,J＝14.4Hz,2H),1.81-1.75(m,1H),1.73-1.65(m,3H),1.63(s,3H),1.62-1.52(m,5H),1.50-1.38(m,3H),1.37-1.27(m,8H),1.05(ddd,J＝17.6,12.4,5.2Hz,2H),0.98-0.95(m,3H),0.93-0.88(m,4H),0.86(s,3H),0.84(d,J＝2.8Hz,1H),0.82-0.78(m,3H).LC-MS[M+H]+＝492.5.
In a second step, p-nitrobenzoic acid (8.20 mg,0.049 mmol) was dissolved in dichloromethane (2 mL) and 2,4, 6-trichlorobenzoyl chloride (15.55 mg,0.064 mmol) and triethylamine (0.020mL, 0.147 mmol) were added at 25 ℃. The reaction solution was stirred at 25℃for 0.5hr.N- (4, 4-difluorodecyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide (175-1) (25 mg,0.049 mmol) and 4-dimethylaminopyridine (1.20 mg, 0.010mmol) were added to the reaction solution. The reaction solution was stirred at 25℃for 16hr. TLC (PE: ea=3:1, uv) showed the reaction was complete. The reaction solution was purified by TLC plate (PE: ea=3:1), and the sample was collected to give 4-nitrobenzoic acid- (2 s,4ar,4bs,8ar,10 ar) -8- {3- [ (4, 4-difluorodecyl) amino ] -3-oxypropylene } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ester (175-2) (18 mg,0.027mmol, 55.71%) as a white solid LC-MS [ m+h ] + = 659.5.
The third step was carried out by subjecting the compound 4-nitrobenzoic acid- (2S, 4aR,4bS,8aR,10 aR) -8- {3- [ (4, 4-difluorodecyl) amino ] -3-oxypropyl } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ester (175-2) (18 mg,0.027 mmol) to SFC resolution purification gave 4-nitrobenzoic acid- (2S, 4aR,4bS,8aR,10 aR) -8- {3- [ (4, 4-difluorodecyl) amino ] -3-oxopropyl } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ester (175-3) (10 mg,0.015mmol, 55.56%) as a white solid. LC-MS [ m+h ] + = 659.5.
The fourth step was to dissolve the compound 4-nitrobenzoic acid- (2S, 4aR,4bS,8aR,10 aR) -8- {3- [ (4, 4-difluorodecyl) amino ] -3-oxoypropyl } -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ester (175-3) (10 mg,0.015 mmol) in methanol (2 mL) followed by addition of potassium carbonate (4.20 mg,0.030 mmol) and reaction at 25℃for 16 hours. LCMS showed complete reaction of starting material and monitored for product formation. The reaction solution was filtered, and the filtrate was subjected to preparative chromatography to give N- (4, 4-difluorodecyl) -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (175) (3.33 mg,0.007mmol, 43.02%) as a white solid .1H NMR(400MHz,CD3OD)δ3.27-3.10(m,3H),2.54-2.44(m,1H),2.31(d,J＝6.4Hz,1H),2.27-2.18(m,2H),2.17-2.08(m,1H),2.07-1.95(m,2H),1.94-1.76(m,6H),1.76-1.57(m,9H),1.50-1.41(m,3H),1.39-1.28(m,7H),1.13-1.00(m,2H),0.96(s,3H),0.91(s,3H),0.90-0.87(m,1H),0.86(s,3H),0.84(d,J＝2.8Hz,1H),0.79(s,3H).LC-MS[M+H]+＝510.5.
Example 174
Example 174 preparation of 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthre ] -N- (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) butanamide
In the first step, the compound { [ (3 aS,3bR,5aR,7S,9aR,9bS,11 aS) -6, 9a,11 a-tetramethyl-1-oxolano [1,2-a ] phenanthren-7-yl ] amino } methano-2-methylpropan-2-yl ester (1.3 g,3.113 mmol) was dissolved in anhydrous dioxane (150 mL), replaced with nitrogen gas by a light reaction apparatus, irradiated with 300W Hg lamp, and reacted at 25℃for 50 minutes. TLC (PE: EA=5:1, phosphomolybdic acid developer) showed that the reaction was completed, the reaction solution was concentrated to give a crude product, and the crude product was separated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give { [ (2S, 4aR,4bS,8aR,10 aR) -8- (2-formylethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-1) (800 mg,1.724mmol, 55.38%) as a white solid .1H NMR(400MHz,CDCl3)δ9.80-9.73(m,1H),3.26(dd,J＝12.4,10.4Hz,1H),2.51-2.29(m,5H),2.17-2.06(m,3H),2.01-1.86(m,3H),1.63(s,3H),1.44(s,9H),1.31-1.21(m,5H),1.14(dd,J＝15.2,7.6Hz,3H),0.95(s,3H),0.90(s,3H),0.73(s,3H).
In the second step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (2-formylethyl) -1, 4a, 7-tetramethyl-1, 2,3, 4-a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-1) (800 mg,1.916 mmol) was dissolved in methanol (10 mL), and then sodium borohydride (144.93 mg,3.831 mmol) was added under nitrogen atmosphere at 0℃to react at 25℃for 2 hours. TLC (PE: ea=3:1, phosphomolybdic acid developer) showed new spot production. The reaction was added to ice water (15 mL), stirred for 10min, extracted with ethyl acetate (15 mL x 3), the organic layers combined and washed with saturated brine (10 mL x 2). The organic layer was dried over sodium sulfate, filtered and concentrated to give the crude product. The crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give { [ (2 s,4ar,4bs,8ar,10 ar) -8- (3-hydroxypropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-2) (600 mg,1.287mmol, 67.18%) as a white solid. LC-MS [ m+h ] + =364.2.
In the third step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-hydroxypropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4-a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-2) (0.6 g,1.430 mmol) was dispersed in methylene chloride (10 mL), N-diisopropylethylamine (0.473 mL, 2.640 mmol), 4-dimethylaminopyridine (0.02 g,0.143 mmol) and t-butyldimethylchlorosilane (0.41 g,2.145 mmol) were added under nitrogen atmosphere at 0℃and the reaction solution was slowly heated to 25℃and stirred for 16h at 25 ℃. TLC (PE: ea=3:1, phosphomolybdic acid developer) showed the reaction was complete. The reaction was quenched with saturated sodium bicarbonate solution (10 mL) and extracted with dichloromethane (3 x 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and the mother liquor concentrated to give crude product. The crude product was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give 4-methylbenzenesulfonic acid-3- [ (4 as,4br,7s,8ar,10 ar) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (174-3) (280 mg, 0.264 mmol, 32.42%) as a white solid. LC-MS [ m+h ] + = 596.4.
In the fourth step, 4-methylbenzenesulfonic acid-3- [ (4 aS,4bR,7S,8aR,10 aR) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propyl ester (174-3) (400 mg,0.697 mmol) was dissolved in dimethyl sulfoxide (10 mL), and sodium cyanide (90 mg,1.836 mmol) and potassium iodide (115.71 mg,0.697 mmol) were added at 25℃and the reaction mixture was stirred at 60℃for 18 hours. TLC plate to detect the completion of the reaction, 10mL of water was added to the reaction solution, followed by extraction with ethyl acetate (3 x 10 mL), washing the organic phase with water (3 x 10 mL), washing with saturated brine (3 x 10 mL), drying over anhydrous sodium sulfate, filtration, spin-drying to give the crude product, which was isolated and purified by column chromatography (petroleum ether: ethyl acetate=1:0) to give { [ (2 s,4ar,4bs,8ar,10 ar) -8- (3-cyanopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4-b,5,6,8a,9,10 a-dodecahydrophenanthre-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-4) (200 mg,0.420mmol, 60.24%) as a pale yellow solid .1H NMR(400MHz,CDCl3)δ3.26(d,J＝14.4Hz,1H),2.38-2.26(m,4H),2.19(d,J＝7.2Hz,2H),2.00-1.93(m,2H),1.75(d,J＝13.6Hz,3H),1.66(s,3H),1.44(s,9H),1.16(s,2H),1.08-1.01(m,6H),0.94(s,2H),0.91-0.88(m,3H),0.80-0.75(m,3H),0.75-0.72(m,3H).
In the fifth step { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-cyanopropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4-a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-4) (200 mg,0.467 mmol) was dissolved in anhydrous dichloromethane (5 mL), diisobutylaluminum hydride (0.933 mL,0.933 mmol) (1 mol/L toluene solution) was added under nitrogen atmosphere at-78℃and the reaction was stirred at-78℃for 2 hours. TLC showed that the reaction had ended, at which temperature 1mol/L potassium sodium tartrate solution (10 mL) and dichloromethane (10 mL) were slowly added dropwise to room temperature and stirred until the two phases became clear, the aqueous phase was extracted with dichloromethane (3X 10 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-formylpropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4a,4-b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-5) (200 mg,0.324mmol, 69.51%) as a white oily crude product which was used directly for the next step without purification .1H NMR(400MHz,CDCl3)δ9.77(d,J＝1.6Hz,1H),3.32-3.22(m,1H),2.48-2.40(m,2H),1.96(s,2H),1.72(d,J＝6.4Hz,4H),1.62(s,3H),1.44(s,9H),1.31-1.19(m,6H),1.09-0.98(m,7H),0.90(s,3H),0.78(s,3H),0.74(s,3H).
In the sixth step, the compound { [ (2S, 4aR,4bS,8aR,10 aR) -8- (3-formylpropyl) -1, 4a, 7-tetramethyl-1, 2,3, 4-a,4b,5,6,8a,9,10 a-dodecahydrophenanthren-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-5) (200 mg,0.463 mmol) was dissolved in t-butanol (5 mL), and an aqueous solution (1 mL) of 2-methylbutan-2-ene (194.99 mg,2.780 mmol), disodium hydrogen phosphate (166.76 mg,1.390 mmol) and sodium chlorite (146.66 mg,1.622 mmol) was added under ice-bath. The reaction was slowly raised to 25℃and stirred at this temperature for 2 hours. TLC (petroleum ether: ethyl acetate=5:1) showed that the reaction was complete. The reaction was quenched by addition of saturated sodium sulfite (10 mL) and extracted with ethyl acetate (3×10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to give a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 5:1) to give 4- [ (4 as,4br,7s,8ar,10 ar) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] butanoic acid (174-6) (110 mg,0.246mmol, 53.04%) as a white solid .1H NMR(400MHz,CDCl3)δ3.34-3.20(m,1H),2.34(td,J＝7.6,4.0Hz,2H),2.09(d,J＝10.4Hz,3H),1.94(d,J＝15.6Hz,3H),1.76-1.67(m,5H),1.60(s,3H),1.44(s,9H),1.31(dd,J＝9.2,3.6Hz,1H),1.18-1.11(m,2H),1.05(d,J＝4.4Hz,4H),0.89(s,3H),0.78(s,3H),0.75(s,1H),0.74(s,3H).
The seventh step was to dissolve the compound 4- [ (4 aS,4bR,7S,8aR,10 aR) -2,4b, 8-tetramethyl-7- ({ [ (2-methylpropan-2-yl) oxy ] carbonyl } amino) -3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] butanoic acid (174-6) (50 mg,0.112 mmol) in acetonitrile (5 mL), then add N, N, N, N-tetramethyl chloroformidine hexafluorophosphate (40.74 mg,0.145 mmol) and N-methylimidazole (32.10 mg,0.391 mmol) and react at 25℃for 0.5 hours. 1-Methylbenzo [ d ] [1,2,3] triazacyclopent-5-amine (33.10 mg,0.223 mmol) was added to the reaction, and reacted at 25℃for 16 hours. TLC (PE: ea=1:1, phosphomolybdic acid developer) showed the reaction was complete. The reaction solution is concentrated to obtain crude products. The crude product was purified by TLC plate to give { [ (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {4- [ (1-methylbenzo [ d ] [1,2,3] triazapentan-5-yl) amino ] -4-oxolanyl } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthre-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-7) (45 mg,0.074mmol, 66.06%) as a white solid. LC-MS [ m+h ] + =578.4.
In an eighth step, starting { [ (2S, 4aR,4bS,8aR,10 aR) -1, 4a, 7-tetramethyl-8- {4- [ (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) amino ] -4-oxobutylene } -1,2,3, 4a,4b,5,6,8a,9,10 a-dodecahydrophenanthre-2-yl ] amino } methanoic acid-2-methylpropan-2-yl ester (174-7) (45 mg,0.078 mmol) was dissolved in 2mL hexafluoroisopropanol. The reaction was carried out at 80℃for 48 hours. LCMS showed complete reaction of starting material and monitored for product formation. The reaction solution was concentrated to obtain a crude product, and the crude product was separated by a liquid chromatography column (hydrochloric acid system) to obtain 4- [ (4 aS,4bR,7S,8aR,10 aR) -7-amino-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- (1-methylbenzo [ d ] [1,2,3] triazacyclopent-5-yl) butyramide (174) (8.75 mg,0.018mmol, 23.08%) as a yellow solid .1H NMR(400MHz,CD3OD)δ8.52-8.47(m,1H),7.81(d,J＝9.2Hz,1H),7.67(dd,J＝9.2,1.6Hz,1H),4.38(s,3H),2.90(dd,J＝9.2,7.2Hz,1H),2.43(t,J＝7.2Hz,2H),2.38-2.19(m,2H),2.19-2.12(m,1H),2.11-2.03(m,1H),2.02-1.95(m,1H),1.94-1.84(m,2H),1.80-1.72(m,3H),1.72-1.66(m,2H),1.63(s,3H),1.58-1.46(m,1H),1.45-1.28(m,1H),1.23-1.10(m,2H),1.05(s,3H),1.03-0.91(m,3H),0.89(d,J＝2.4Hz,6H).LC-MS[M+H]+＝478.4.
Example 176
Example 176 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (2-methoxyphenyl) heptyl ] propanamide
In the first step 1, 7-dibromoheptane (55.72 g,205.161 mmol) was dissolved in acetone (400 mL) at 25℃and potassium 1, 3-dioxy-2, 3-dihydro-1H-isoindol-2-yl anion (20 g,102.581 mmol) was added followed by stirring at 80℃for 16 hours. TLC plate (petroleum ether: ethyl acetate=5:1, uv) indicated the starting material was exhausted and new spots of reduced polarity were produced, the reaction was filtered through celite and the filtrate was concentrated and spun dry to give crude product. The crude product was isolated and purified by column chromatography on silica gel (petroleum ether: ethyl acetate=6:1) to give 2- (7-bromoheptyl) isoindole-1, 3-dione (176-1) (16 g,48.362mmol, 47.15%) as a colourless oil .1H NMR(400MHz,CDCl3)δ7.86-7.80(m,2H),7.74-7.66(m,2H),3.67(t,J＝7.2Hz,2H),3.38(t,J＝6.8Hz,2H),1.83(q,J＝7.2Hz,2H),1.70-1.65(m,2H),1.46-1.38(m,2H),1.38-1.32(m,4H).
In a second step, compound 2- (7-bromoheptyl) isoindole-1, 3-dione (176-1) (2 g,5.860 mmol) was dissolved in N, N-dimethylacetamide (20 mL), followed by the addition of 1-iodo-2-methoxybenzene (1.37 g,5.860 mmol), sodium iodide (0.22 g, 1.460 mmol), zinc powder (0.96 g,14.651 mmol), nickel (II) chloride glycol dimethyl ether complex (0.13 g,0.586 mmol) and 2-amidinopyridine hydrochloride (0.09 g,0.586 mmol). The flask was purged with nitrogen for 5 minutes to change the color from blue to red, and the solution was changed to black after reaction at 60℃under nitrogen for 16 hours. TLC (PE: ea=20:1, uv) showed the starting material was essentially completely reacted and a new spot was generated. The reaction solution was filtered through celite, the filter cake was washed with ethyl acetate (30 ml×2), the filtrate was concentrated, then saturated ammonium chloride (40 mL) was added to wash, extracted with ethyl acetate (30 ml×2), and the combined organic layers were washed with saturated brine (50 ml×2). The organic layer was dried over Na 2 SO4, filtered and concentrated to give the crude product. The crude product was purified by flash chromatography (PE: etoac=30:1) and the sample was collected to give 2- [7- (2-methoxyphenyl) heptyl ] isoindole-1, 3-dione (176-2) (570 mg, 1.540 mmol, 26.29%) as a yellow oil .1H NMR(400MHz,CDCl3)δ7.87-7.81(m,2H),7.74-7.66(m,2H),7.19-7.07(m,2H),6.94-6.78(m,2H),3.81(s,3H),3.67(t,J＝7.2Hz,2H),2.69-2.49(m,2H),1.70-1.64(m,2H),1.60-1.51(m,2H),1.35(s,6H).
In the third step, the starting material 2- [7- (2-methoxyphenyl) heptyl ] isoindole-1, 3-dione (176-2) (294 mg,0.846 mmol) was dissolved in absolute ethanol (5 mL), and hydrazine hydrate (0.386 mL,6.352 mmol) (80%) was added under ice-bath and the reaction was stirred at 25℃for 1 hour. TLC (Petroleum ether: ethyl acetate=10:1) showed that the starting material had reacted completely with the formation of a more polar spot, the reaction solution was filtered, the cake was washed with ethyl acetate, a white solid was produced during concentration of the filtrate under reduced pressure, and then filtration was repeated until concentration under reduced pressure gave 7- (2-methoxyphenyl) hept-1-amine (176-3) (163 mg,0.589mmol, 87.18%) as a yellow transparent oily product, which was crude and was used directly in the next step .1H NMR(400MHz,CDCl3)δ7.19-7.12(m,2H),6.90-6.83(m,2H),3.82(s,3H),2.69(t,J＝7.2Hz,2H),2.62-2.58(m,2H),1.58(br d,J＝7.2Hz,2H),1.45(td,J＝13.6,6.8Hz,2H),1.38-1.33(m,6H).
In the fourth step, 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecylphenanthryl ] propionic acid (III) (20 mg,0.054 mmol) (90%) was dissolved in methylene chloride (3 mL), diisopropylethylamine (0.018 mL,0.108 mmol) and O- (7-azabenzotriazol-1-YL) -N, N, N, N-tetramethylurea hexafluorophosphine salt (30.69 mg,0.081 mmol) were added, the reaction solution was stirred for 30 minutes, and then 7- (2-methoxyphenyl) hept-1-amine (176-3) (17.87 mg,0.065 mmol) was added to the reaction solution, and the reaction solution was stirred at 25℃for 18 hours. LCMS showed the reaction was complete. The reaction mixture was dried under reduced pressure to give a crude product, which was isolated and purified by TLC (Petroleum ether: ethyl acetate=3:1) to give 25mg of the product as a yellow oil, which was further isolated and purified by HPLC (HCl) to give 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] -N- [7- (2-methoxyphenyl) heptyl ] propanamide (176) (14.45 mg,0.027mmol, 49.62%) as a pale yellow oil .1H NMR(500MHz,CD3OD)δ7.94-7.86(m,1H),7.17-7.02(m,2H),6.93-6.74(m,2H),3.80(s,3H),3.22-3.06(m,3H),2.58(t,J＝7.5Hz,2H),2.54-2.37(m,1H),2.36-2.16(m,3H),2.15-1.95(m,3H),1.94-1.81(m,2H),1.78-1.63(m,5H),1.60-1.52(m,3H),1.50-1.42(m,3H),1.33(s,7H),1.10-1.01(m,2H),0.97-0.94(m,3H),0.91-0.82(m,6H),0.80-0.78(m,3H).LC-MS[M+H]+＝538.5.
Example 177
Example 177 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (4-methoxyphenyl) heptyl ] propanamide
Reference example 176 conversion of 1-iodo-2-methoxybenzene to 1-iodo-4-methoxybenzene in the second step, followed by the same operation in the three subsequent steps, gave 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (4-methoxyphenyl) heptyl ] propanamide (177).1H NMR(500MHz,CD3OD)δ7.06(d,J＝8.8Hz,2H),6.80(d,J＝8.8Hz,2H),3.75(s,3H),3.22-3.08(m,3H),2.56-2.42(m,3H),2.29(s,1H),2.26-2.17(m,2H),2.16-2.07(m,1H),2.06-1.95(m,2H),1.94-1.82(m,2H),1.81-1.75(m,1H),1.72-1.65(m,2H),1.62(s,3H),1.60-1.54(m,2H),1.52-1.41(m,3H),1.37-1.28(m,7H),1.12-0.99(m,2H),0.98-0.94(m,3H),0.93-0.86(m,2H),0.85(s,3H),0.83(s,1H),0.81-0.78(m,3H).LC-MS[M+H]+＝538.5.
Example 178
Example 178 preparation of 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (3-methoxyphenyl) heptyl ] propanamide
Reference example 176 conversion of 1-iodo-2-methoxybenzene to 1-iodo-3-methoxybenzene in the second step, followed by the same operation in the three subsequent steps, gave 3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] -N- [7- (3-methoxyphenyl) heptyl ] propanamide (178).1H NMR(500MHz,CD3OD)δ7.18-7.14(m,1H),6.77-6.70(m,3H),3.78(s,3H),3.22-3.07(m,3H),2.58(t,J＝7.5Hz,2H),2.53-2.45(m,1H),2.34-2.27(m,1H),2.26-2.19(m,2H),2.16-2.08(m,1H),2.07-1.96(m,2H),1.95-1.85(m,2H),1.82-1.72(m,2H),1.72-1.58(m,9H),1.54-1.44(m,3H),1.43-1.28(m,8H),1.07(d,J＝3.2Hz,2H),0.99-0.96(m,3H),0.88(s,1H),0.86(s,3H),0.82-0.79(m,3H).LC-MS[M+H]+＝538.5.
Example 179
Example 179 preparation of N- [7- (2-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
Reference example 176 conversion of 1-iodo-2-methoxybenzene to 1-iodo-2-cyanobenzene in the second step, followed by the same operation in the three subsequent steps, gave N- [7- (2-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (179).1H NMR(400MHz,CDCl3)δ7.62-7.60(m,1H),7.53-7.50(m,1H),7.33-7.28(m,2H),5.53-5.50(m,1H),3.25-3.21(m,3H),2.85-2.82(m,2H),2.49-2.40(m,1H),2.38-2.30(m,1H),2.28-2.22(m,1H),2.18-2.10(m,2H),2.04-1.96(m,2H),1.93-1.87(m,2H),1.78-1.74(m,1H),1.74-1.67(m,5H),1.60(m,2H),1.50(m,2H),1.40-1.34(m,7H),1.27(m,1H),1.09-1.02(m,2H),0.97(s,3H),0.94-0.90(m,1H),0.89-0.85(m,2H),0.83-0.81(m,4H),0.79(s,3H).LC-MS[M+H]+＝533.3.
Example 180
Example 180 preparation of N- [7- (3-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
Reference example 176 conversion of 1-iodo-2-methoxybenzene to 1-iodo-3-cyanobenzene in the second step, followed by the same operation in the three subsequent steps, gave N- [7- (3-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (180).1H NMR(400MHz,CDCl3)δ7.50-7.45(m,2H),7.42-7.35(m,2H),5.50-5.43(m,1H),3.25-3.19(m,3H),2.65-2.61(m,2H),2.47-2.31(m,2H),2.25-2.11(m,3H),2.00-1.88(m,3H),1.76-1.67(m,4H),1.60-1.57(m,2H),1.50-1.46(m,2H),1.42-1.35(m,2H),1.33(m,5H),1.26(s,3H),1.06-1.00(m,2H),0.98-0.95(m,3H),0.92-0.85(m,4H),0.82-0.80(m,4H),0.79(s,3H).LC-MS[M+H]+＝533.3.
Example 181
Example 181 preparation of N- [7- (4-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
Reference example 176 conversion of 1-iodo-2-methoxybenzene to 1-iodo-4-cyanobenzene in the second step, followed by the same operation in the three subsequent steps, gave N- [7- (4-cyanophenyl) heptyl ] -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthreyl ] propanamide (181).1H NMR(400MHz,CD3OD)δ7.93(s,1H),7.62(d,J＝8.0Hz,2H),7.37(d,J＝8.0Hz,2H),3.23-3.09(m,3H),2.70(t,J＝7.6Hz,2H),2.52-2.41(m,1H),2.32-2.16(m,3H),2.06-1.96(m,2H),1.94-1.84(m,2H),1.81-1.73(m,2H),1.62(s,9H),1.49-1.44(m,2H),1.35(s,6H),1.29(d,J＝3.6Hz,2H),1.09-1.01(m,2H),0.96(s,3H),0.91-0.87(m,1H),0.85(s,3H),0.79(s,3H).LC-MS[M+H]+＝533.3.
Example 182
Example 182 preparation of N- {1- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-yl } -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrene ] propanamide
In a first step, compound 2- (2, 5-difluorophenyl) ethan-1-ol (4.4 g,27.82 mmol) was dissolved in dichloromethane (50 mL) and 2, 6-lutidine (11.92 g,111.29mmol,12.96mL,4 eq.) and methanesulfonic anhydride (9.69 g,55.65mmol,2 eq.) were added at 25℃the reaction was reacted at 0℃for 1hr, then at 25℃for 12hr. LCMS showed complete reaction of starting material with product. TLC showed new spots generated. The reaction was quenched with 50mL of water, extracted with ethyl acetate (30 mL x 2), and the combined organic layers were washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated to give crude methanesulfonic acid-2- (2, 5-difluorophenyl) ethyl ester (182-1) (6.57 g,27.81mmol,100.00% yieldd) as a red oil, which was used directly in the next step. LC-MS [ m+na ] + =259.2.
In a second step, the compound methanesulfonic acid-2- (2, 5-difluorophenyl) ethyl ester (182-1) (6.57 g,27.81mmol,1 eq.) was dissolved in acetonitrile (120 mL), followed by the addition of potassium carbonate (15.38 g,111.25mmol,4 eq.) and 3-N-BOC-aminoazetidine (4.79 g,27.81 mmol.) 16hr at 80℃LCMS showed complete reaction of the starting materials with the formation of the product. TLC showed new spots generated. The reaction was quenched with 50mL of water, extracted with ethyl acetate (50 mL x 2), the combined organic layers were washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated, and purified by column chromatography (petroleum ether: ethyl acetate=1:0 to 3:1) to give ({ 1- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-yl } amino) methanoic acid-2-methylpropan-2-yl ester (182-2) (5.21 g,16.68mmol,59.97% yieldd) as a yellow oil. LC-MS [ m+h ] + =313.2.
In the third step, 2-methylpropan-2-yl ester (182-2) (200 mg,0.640 mmol) of the compound ({ 1- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-yl } amino) methanoate was dissolved in dichloromethane (6 mL), trifluoroacetic acid (2 mL) was added, and the reaction was carried out at 25℃for 2hr.LCMS, showing that the starting material was complete and the product was formed. The reaction solution was concentrated to give crude 1- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-amine (182-3) (200 mg,0.552mmol, 86.20%) as a red solid, which was used directly in the next step. LC-MS [ m+h ] + =213.2.
Fourth step referring to the fourth step in example 176, 7- (2-methoxyphenyl) hept-1-amine was changed to- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-amine to give N- {1- [2- (2, 5-difluorophenyl) ethyl ] azetidin-3-yl } -3- [ (4 aS,4bR,7S,8aR,10 aR) -7-hydroxy-2, 4b, 8-tetramethyl-3, 4a,4b,5,6,7, 8a,9,10 a-dodecahydrophenanthrenyl ] propionamide (182).1H NMR(400MHz,CD3OD)δ7.12-7.03(m,2H),7.01-6.94(m,1H),4.46-4.36(m,1H),3.71-3.65(m,2H),3.18(dd,J＝10.8,5.6Hz,1H),3.04-2.94(m,2H),2.73(qd,J＝11.6,5.6Hz,4H),2.53-2.41(m,1H),2.37-2.27(m,1H),2.26-2.22(m,1H),2.18-2.09(m,1H),2.04(d,J＝10.8Hz,2H),1.97-1.83(m,2H),1.82-1.77(m,1H),1.75-1.69(m,2H),1.68-1.61(m,5H),1.45(dt,J＝12.8,9.2Hz,1H),1.32(s,1H),1.06(dt,J＝12.4,5.2Hz,2H),1.00-0.96(m,3H),0.93-0.89(m,1H),0.88-0.84(m,4H),0.83-0.80(m,3H).LC-MS[M+H]+＝529.4.
Effect example 1: inhibition assay for Gli Gene luciferase reporter
NIH-3T3/Gli-luc/12 cells (constructed cell lines stably expressing Gli-luc reporters and zsGreen) were seeded at a cell mass of 2X 10 4 per well in 96-well plates and incubated with DMEM medium (containing 10% neonatal bovine serum (sigma B7447)) in a 5% CO2 incubator at 37 ℃. After 48h, the original medium was aspirated and replaced with DMEM medium for 6 h. Starvation medium (containing Shh-N conditional medium) was then changed to contain different concentrations of the compound. The highest concentration of compound 1000nM, 3-fold dilution, total 8 concentrations. After 30 hours of incubation, the cells were washed once with PBS Buffer, then 50. Mu.L of the prepared 1X PASSIVE LYSIS Buffer (Promega E1941) was added to each well, and the lysed cells were assayed for self-luminescence using the firefly luciferase kit (Promega E1501) while simultaneously assayed for EGFP (excitation light 485, reception light 528) as the inhibition capacity of the Hh signaling pathway by each test compound using the ratio of the self-luminescence reading value to the EGFP reading value, i.e., the unit fluorescence value.
The experimental results are shown in the following table:
Remarks: distinguishing the activity range of the compound according to IC 50, wherein A is less than or equal to 0.1 mu M;0.1 mu M < B.ltoreq.1 mu M; c > 1. Mu.M.
The results show that the compound provided by the invention has good inhibition capability on the Hh signal path.
Effect example 2: inhibition experiments of Gli Gene overexpressing SMO mutant
NIH3T 3-cells stably expressing the human SMO mutant (D437H) with knockout of endogenous SMO were cultured in DMEM medium (containing 10% neonatal bovine serum (sigma B7447)) at 37 ℃ in a 5% co2 incubator. After 48h, the original medium was aspirated and replaced with DMEM medium for 6 h. Starvation medium (containing Shh-N conditional medium) was then changed to contain different concentrations of the compound. The highest concentration of compound 1000nM, 3-fold dilution, total 8 concentrations. After 30 hours of incubation, mRNA extraction, cDNA reverse transcription, and qPCR were performed to detect Gli gene expression.
The compound has good inhibition capability on SMO drug-resistant mutation.
Effect example 3: inhibition experiment of Gli Gene luciferase reporter Gene of SMO mutant
Endogenous SMO-knocked out NIH-3T3 cells were seeded at 2.5×10 4 cells per well in 24-well plates and incubated with DMEM medium (containing 10% neonatal bovine serum (sigma B7447), diabody) at 37 ℃ in a 5% co2 incubator. SMO-D473H, SMO-E518K mutant plasmids pGLIBS and pEGFP-N1 (total transfection amount 0.25. Mu.g, SMO 0.125. Mu.g, pGLIBS and pEGFP co-transfected at 1:20) were transfected into cells using lipo 2000. After 6-8 hours, the liquid is changed. The original medium was aspirated off and replaced with DMEM medium starved for 6 hours. Starvation medium (containing Shh-N conditional medium) was then changed to contain different concentrations of the compound. The Hh signaling pathway is maximally activated upon stimulation by Shh-N. After 48 hours of culture, the cells are washed once by PBS Buffer solution, then 100 mu L of prepared 1X PASSIVE LYSIS Buffer (Promega E1941) is added into each hole, the cells are placed in a refrigerator at the temperature of minus 80 ℃ for freezing for 5 minutes (or overnight), and after thawing at room temperature, the cells are placed on a horizontal shaker for shaking and mixing for 30 minutes at 800rpm, so that the purpose of fully lysing the cells is achieved. The self-luminescence value of the lysed cells is measured by a firefly luciferase kit (Promega E1501), the EGFP value (excitation light is 485 and receiving light is 528) is measured by a Cytation multifunctional enzyme-labeled instrument, and the ratio of the self-luminescence reading value to the EGFP reading value, namely the unit fluorescence value, is used as the inhibition capability of each test compound on the Hh signal path.
The compound has good inhibition capability on SMO drug-resistant mutation.
Effect example 4: liver microsome metabolic stability experiment
PBS solution (100 mM), mgCl2 solution (100 mM) and NADPH solution (20 mM) were prepared separately, and then compound and testosterone stock solution were prepared separately with dimethyl sulfoxide (DMSO), diluted to 100. Mu.M with methanol for sample incubation, and stored at-10-30 ℃. 12.5. Mu.L of liver microsomes (rat/mouse/dog/monkey/human), 432.5. Mu.L of PBS solution (100 mM), 25. Mu.L of NADPH solution (20 mM) and 25. Mu.L of MgCl2 solution (100 mM) are added to a 96-well plate and mixed uniformly, and then preincubated at 37℃for 5min; adding 5 mu L of substrate solution to start the reaction; and respectively taking 50 mu L of incubation samples at each set time point of 0, 5, 15, 30, 45 and 60min (negative group is 0 and 60 min), placing the incubation samples in a termination plate added with 100 mu L of ice termination solution, inactivating by vortex for 1min, and preserving at-60 to-90 ℃ to be tested. Sample analysis of test and control compounds testosterone was performed using LC-MS/MS methods. Chromatographic peaks were integrated, calculated and processed using analysis software. The test drug and the control compound testosterone were both calculated as peak area ratios using semi-quantitative analysis.
The compound of the invention has good metabolism stability of liver microsomes.

Claims (20)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:
Wherein,
R 1 is-OH, -NH 2 or-NR 1-1SO2R1-2; or R 1 is bound to a carbon atom to form
R 1-1 and R 1-2 are independently H or C 1-C6 alkyl;
l 1 is- (CH 2) n-where n is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -are replaced by-CHR L-1 -or-CR L-1RL-1 -;
R L-1 is independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-C(＝O)NR2-1-、-NR2-1C(＝O)-、-NR2-1C(＝O)NR2-1-、-C(＝O)-、-C(＝O)O-、-C(＝O)NR2-1NR2-1C(＝O)-、-NR2-1C(＝O)O-、-SO2NR2-1- or 5-10 membered heteroarylene, in L 2, the hetero atom in the heteroarylene is one, two or three selected from N, O and S, and the number of the hetero atoms is 1, 2 or 3;
r L-2' is OH; r L-2 is H or C 1-C6 alkyl;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1;
R 2-1-1 is independently OH, COOH or C 1-C6 alkoxy;
L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, -CHR L-3-、-NRL-4 -, -O-, -S-or 3-6 membered heterocycloalkylene; in L 3, in the heterocycloalkylene group, the type of the heteroatom is one, two or three selected from N, O and S, and the number of the heteroatom is 1, 2 or 3;
R L-3 is independently C 1-C6 alkyl or C 1-C6 alkoxy;
R L-4 is independently H or C 1-C6 alkyl, C 1-C6 alkoxy, C 1-C6 alkyl substituted with 1, 2 or 3R 2-2;
R 2-2 is OH, NR 2-2-1R2-2-1, halogen, C 1-C6 alkoxy, 5-6 membered heteroaryl, -C (=O) NR 2-2-1R2-2-1、-NR2-2-1C(＝O)-C1-C6 alkyl, -NR 2-2-1C(＝O)NR2-2-1R2-2-1、-C(＝O)-C1-C6 alkyl, -C (=O) OR 2-2-1、-C(＝O)NR2-2-1NR2-2-1C(＝O)C1-C6 alkyl OR-SO 2NR2-2-1R2-2-1;R2-2, wherein the hetero atoms are selected from one, two OR three of N, O and S, and the number of the hetero atoms is 1,2 OR 3;
R 2-2-1 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4, wherein in A, the type of heteroatom is selected from one, two or three of N, O and S, and the number of heteroatoms is 1, 2, 3 or 4; in the A, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1, 2 or 3;
R 3-1 is independently oxo, CN, NR 3-1-1R3-1-2、COOR3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1, 2 or 3R 3-1-4, wherein in the heterocycloalkyl, the type of heteroatom is selected from one, two or three of N, O and S, and the number of the heteroatom is 1, 2 or 3;
r 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl;
R 3-1-4 is independently halogen or 5-10 membered heteroaryl, wherein in the heteroaryl, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2, 3 or 4;
R 3-2 is independently halogen, (c=o) NR 3-2-3R3-2-4、C1-C6 alkyl, C 1-C6 alkoxy, -NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
wherein the compound represented by formula (I) is not the following compound or stereoisomer thereof:
2. A compound of formula (I) or a pharmaceutically acceptable salt thereof:
Wherein,
R 1 is-OH, NH 2 or-NR 1-1SO2R1-2; or R 1 is bound to a carbon atom to form
R 1-1 and R 1-2 are independently H or C 1-C6 alkyl;
l 1 is- (CH 2) n-where n is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -are replaced by-CHR L-1 -or-CR L-1RL-1 -;
R L-1 is independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-C(＝O)NR2-1-、-NR2-1C(＝O)-、-NR2-1C(＝O)NR2-1-、-C(＝O)-、-C(＝O)O-、-C(＝O)NR2-1NR2-1C(＝O)-、-NR2-1C(＝O)O-、-SO2NR2-1- or 5-6 membered heteroarylene, in L 2, the hetero atom in the heteroarylene is one, two or three selected from N, O and S, and the number of the hetero atoms is 1,2 or 3;
r L-2' is OH; r L-2 is H or C 1-C6 alkyl;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1;
R 2-1-1 is independently OH, COOH or C 1-C6 alkoxy;
L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, -CHR L-3-、-NRL-4 -, -O-, -S-or 3-6 membered heterocycloalkylene; in L 3, in the heterocycloalkylene group, the type of the heteroatom is one, two or three selected from N, O and S, and the number of the heteroatom is 1, 2 or 3;
R L-3 is independently C 1-C6 alkyl or C 1-C6 alkoxy;
R L-4 is independently H or C 1-C6 alkyl, C 1-C6 alkoxy, C 1-C6 alkyl substituted with 1, 2 or 3R 2-2;
R 2-2 is OH, NR 2-2-1R2-2-1, halogen, C 1-C6 alkoxy, 5-6 membered heteroaryl, -C (=O) NR 2-2-1R2-2-1、-NR2-2-1C(＝O)-C1-C6 alkyl, -NR 2-2-1C(＝O)NR2-2-1R2-2-1、-C(＝O)-C1-C6 alkyl, -C (=O) OR 2-2-1、-C(＝O)NR2-2-1NR2-2-1C(＝O)C1-C6 alkyl OR-SO 2NR2-2-1R2-2-1;R2-2, wherein the hetero atoms are selected from one, two OR three of N, O and S, and the number of the hetero atoms is 1,2 OR 3;
R 2-2-1 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4, wherein in A, the type of heteroatom is selected from one, two or three of N, O and S, and the number of heteroatoms is 1, 2, 3 or 4; in the A, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1, 2 or 3;
R 3-1 is independently CN, NR 3-1-1R3-1-2、COOR3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1,2 or 3R 3-1-4, wherein the hetero atoms are selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2 or 3;
r 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl;
R 3-1-4 is independently halogen or 5-10 membered heteroaryl, wherein in the heteroaryl, the hetero atom is selected from one, two or three of N, O and S, and the number of the hetero atoms is 1,2, 3 or 4;
R 3-2 is independently halogen, (c=o) NR 3-2-3R3-2-4、C1-C6 alkyl, C 1-C6 alkoxy, -NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Wherein the compound shown in the formula (I) is not the following compound:
3. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein when L 2 is a 5-10 membered heteroarylene group, the 5-10 membered heteroarylene group is a 5-6 membered monocyclic heteroarylene group or a 9-10 membered bicyclic heteroarylene group, such as a 5-6 membered heteroarylene group; the hetero atom in the heteroarylene group is selected from one or both of N and O, and the number of hetero atoms is 2 or 3, for example, triazolylene, oxadiazolylene or benzimidazolylene, further for example, 1,2, 3-triazolylene, 1,3, 4-oxadiazolylene, 1,2, 4-oxadiazolylene or benzimidazolylene, preferably
Further preferred isWherein the j side is connected with L 1.
4. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 which meets one or more of the following conditions:
(1)R1-1、R1-2、RL-1、RL-2、R2-1、RL-3、RL-4、R2-2、R3-1、R3-3、R2-2-1、R3-1-1、R3-1-2、R3-1-3、R3-2、R3-2-1、R3-2-2、R3-2-3、R3-2-4、 And R 3-4, each "C 1-C6 alkyl" is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl, preferably methyl, ethyl or tert-butyl;
(2) When L 2 is a 5-to 6-membered heteroarylene group, the 5-to 6-membered heteroarylene group is a 5-membered heteroarylene group having a heteroatom selected from one or both of N and O and having a heteroatom number of 2 or 3, such as a triazolylene group or an oxadiazolylene group, further such as a1, 2, 3-triazolylene group, a1, 3, 4-oxadiazolylene group or a1, 2, 4-oxadiazolylene group, preferably Further preferred isWherein the j side is connected with L 1;
(3) In R 2-1-1、RL-3、RL-4、R2-2 and R 3-2, said C 1-C6 alkoxy is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy, preferably methoxy;
(4) In L 3, the 3-6 membered heterocycloalkylene group is one or two of N and O, and the 5-6 membered heterocycloalkylene group having 1 or 2 hetero atoms, such as piperidylene, tetrahydropyranyl, piperazinylene or morpholinylene, further such as piperidylene, preferably
(5) In A, each "C 6-C10 aryl" is independently phenyl or naphthyl;
(6) In A, each "5-10 membered heteroaryl" is independently a 5-6 membered heteroaryl or a 9-10 membered bicyclic heteroaryl, such as pyridinyl, oxadiazolyl, azaindolyl, benzimidazolyl, indolyl, benzothiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, indazolyl, benzotriazolyl, indolinyl, benzomorpholinyl, benzoxadiazolyl, quinazolinyl, quinolinyl, isoquinolinyl, 4,5,6, 7-tetrahydro-1H-benzimidazolyl or 2, 3-dihydrobenzimidazolyl, more such as
(7) In R 3-1-4, each "5-10 membered heteroaryl" is independently a 5-6 membered heteroaryl or a 9-10 membered bicyclic heteroaryl, such as pyridinyl, oxadiazolyl, azaindolyl, benzimidazolyl, indolyl, benzothiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, indazolyl, benzotriazolyl, indolinyl, benzomorpholinyl, benzoxadiazolyl, quinazolinyl, quinolinyl, or isoquinolinyl, more such as
(8) Each "C 3-C6 cycloalkyl" in a and R 3-1 is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl;
(9) In A and R 3-1, each "3-6 membered heterocycloalkyl" is independently a heteroatom selected from one or both of N and O, a 5-6 membered heterocycloalkyl having 1 or 2 heteroatoms, such as piperidinyl, tetrahydropyranyl, morpholinyl or piperazinyl, further such as
(10) In R 2-2、R3-1、R3-1-4 and R 3-2, each "halogen" may independently be F, cl, br, or I, such as F or Cl;
(11) In R 2-2, the "5-6 membered heteroaryl" is one or two of N and O, and the number of the heteroatoms is 1 or 2, such as pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyridazinyl or pyrazinyl.
5. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, which meets one or more of the following conditions:
(1) R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
(2) L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NR2-1NR2-1(C＝O)-、-NR2-1(C＝O)O- or a 5-to 10-membered heteroarylene.
6. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 which meets one or more of the following conditions:
(1) R 1 is-OH or-NR 1-1SO2R1-2;
(2) R 1-1 is H;
(3) R 1-2 is C 1-C6 alkyl;
(4) R 3-1 is independently CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
(5) R 3-1-1 and R 3-1-2 are independently H;
(6) R 3-1-3 is H or C 1-C6 alkyl;
(7) R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
(8) R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
(9) When R L-4 is C 1-C6 alkyl substituted with 1,2 or 3R 2-2, said R 2-2 is at the end of C 1-C6 alkyl, preferably R 2-2 is 1;
(10) 0, 1,2 or 3-CH 2 -in L 1 are replaced with CHR L-1;
(11) n is 2, 3 or 4, preferably 2 or 3;
(12) R 2-1 is H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3C 1-C6 alkoxy groups;
(13) 0,1, 2 or 3-CH 2 -in L 3 are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene;
(14) m is 1, 2, 3, 4, 5, 6, 7, 8,9, 10 or 11;
(15) R L-4 is H or C 1-C6 alkyl.
7. The compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1,
A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3-4;
R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 3-1-4, R 3-1-4 is independently halogen or 5-10 membered heteroaryl;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Preferably, A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1 or C 6-C10 aryl substituted with 1,2 or 3R 3-2;
R 3-1 is independently oxo, CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 3-1-4, R 3-1-4 is independently halogen or 5-10 membered heteroaryl;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl.
8. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 which meets one or more of the following conditions:
(1) R 3-1 is independently CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted by 1,2 or 3R 3-1-4, R 3-1-1 and R 3-1-2 are independently H, R 3-1-3 is H or C 1-C6 alkyl;
(2) R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4,R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 is independently C 1-C6 alkyl.
9. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2 which meets one or more of the following conditions:
(1) R 1 is-OH or-NR 1-1SO2R1-2,R1-1 is H, R 1-2 is C 1-C6 alkyl;
(2) L 1 is- (CH 2) n-where n is an integer from 1 to 12, where 0, 1,2 or 3-CH 2 -are replaced by CHR L-1;RL-1 being C 1-C6 alkyl; preferably, n is 2,3 or 4; more preferably, n is 2 or 3;
(3) L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NR2-1NR2-1(C＝O)-、-NR2-1(C＝O)O- or a 5-6 membered heteroarylene; preferably, L 2 is -CH(OH)-、-NH-、-(C＝O)NR2-1-、-NH(C＝O)-、-NH(C＝O)NR2-1-、-(C＝O)-、-(C＝O)O-、-(C＝O)NHNH(C＝O)-、-NH(C＝O)O- or a 5-6 membered heteroarylene; more preferably, L 2 is- (c=o) NR 2-1、-NH(C＝O)-、-NH(C＝O)NH-、-NH-,R2-1 is H, C 1-C6 alkyl or C 1-C6 alkyl substituted by 1, 2 or 3C 1-C6 alkoxy groups;
(4) L 3 is a bond or- (CH 2) m-, m is an integer from 1 to 12, 0, 1, 2 or 3-CH 2 -in- (CH 2) m-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is H or C 1-C6 alkyl; preferably, L 3 is a bond or- (CH 2) m-, m is 1, 2,3, 4,5, 6,7,8, 9, 10 or 11, wherein 0 or 1-CH 2 -in- (CH 2) m-is replaced by-CH=CH-, NH, O or 3-6 membered heterocycloalkylene, when 0 or 1-CH 2 -in- (CH 2) m-is replaced with a 3-6 membered heterocycloalkylene, the 3-6 membered heterocycloalkylene is linked to L 2;
(5) A is H, CN, -NR 3-3R3-3、-OR3-3、-C(＝O)OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4;
r 3-1 is independently CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
Preferably, A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1 or C 6-C10 aryl substituted with 1,2 or 3R 3-2;
r 3-1 is independently CN, COOR 3-1-3、CONR3-1-1R3-1-2, halogen, C 1-C6 alkyl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
R 3-1-1 and R 3-1-2 are independently H;
r 3-1-3 is H or C 1-C6 alkyl;
R 3-2 is independently halogen, (c=o) NH 2、C1-C6 alkyl, NR 3-2-1(S＝O)2R3-2-2 or NR 3-2-3R3-2-4;
R 3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl.
10. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, which meets one or more of the following conditions:
(1) R 1 is-OH or-NHSO 2CH3;
(2) L 1 is
Preferably is
Wherein side c is attached to L 2;
(3) L 2 is
Preferably is
Wherein the d side is connected with L 1; (4) L 3 is a connecting bond,
Preferably a connecting bond,
Wherein the e side is connected with A;
(5) A is H,
11. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, which meets one or more of the following conditions:
(1) -L 1-L2-L3 -is:
-in L 1-L2-L3 -, L 1 is- (CH 2) n-, n is 2, 3 or 4, wherein 0, 1,2 or 3-CH 2 -are replaced by CHR L-1 or-CR L -1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-to 10-membered heteroarylene group;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
L 3 is a bond;
(2) A is-NR 3-3R3-3、C6-C10 aryl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1, C 6-C10 aryl substituted with 1, 2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4;
R 3-1 is independently oxo, CN, COOR 3-1-3、C1-C6 alkyl, halogen, CONR 3-1-1R3-1-2、C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4;
R 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl; r 3-1-4 is independently halogen or 5-10 membered heteroaryl;
r 3-2 is independently OH, NR 3-2-1(S＝O)2R3-2-2、NR3-2-3R3-2-4;R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
r 3-4 is independently C 1-C6 alkyl.
12. A compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 or 2, wherein-L 1-L2-L3 -is any one of the following:
i: -in L 1-L2-L3 -, L 1 is- (CH 2) n-, n is 2,3 or 4, wherein 0 or 1 of- (CH 2) n-CH 2 -are replaced by CHR L-1 -or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-NR2-1(C＝O)O- or 5-6 membered subunit
Heteroaryl; r L-2' is OH; r L-2 is H or C 1-C6 alkyl; r 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 2-1-1; r 2-1-1 is independently COOH or C 1-C6 alkoxy;
L 3 is- (CH 2) m-, m is an integer from 1 to 12, wherein, 0,1, 2 or 3-CH 2 -in- (CH 2) n-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is independently H or C 1-C6 alkyl;
ii: -in L 1-L2-L3 -, L 1 is- (CH 2) n-, n is 2, 3 or 4, wherein 0,1,2 or 3-CH 2 -are replaced by CHR L-1 or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-6 membered heteroarylene group;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
L 3 is a bond;
Preferably, -L 1-L2-L3 -A is any of the following:
I: -in L 1-L2-L3 -a, L 1 is- (CH 2) n-, n is 2, 3 or 4, wherein 0 or 1 of- (CH 2) n-CH 2 -are replaced by CHR L-1 -or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
L 2 is -CHRL-2'-、-NRL-2-、-(C＝O)NR2-1、-NR2-1(C＝O)NR2-1-、-(C＝O)-、-NR2-1(C＝O)O- or 5-6 membered subunit
Heteroaryl; r L-2' is OH; r L-2 is H or C 1-C6 alkyl; r 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1, 2 or 3R 2-1-1; r 2-1-1 is independently COOH or C 1-C6 alkoxy;
L 3 is- (CH 2) m-, m is an integer from 1 to 12, wherein, 0,1, 2 or 3-CH 2 -in- (CH 2) n-are replaced by-ch=ch-, NR L-4, O or 3-6 membered heterocycloalkylene; r L-4 is independently H or C 1-C6 alkyl;
A is H, CN, -NR 3-3R3-3、-OR3-3、C6-C10 aryl, 5-10 membered heteroaryl, C 3-C6 cycloalkyl, 3-6 membered heterocycloalkyl, C 6-C10 aryl substituted with 1, 2 or 3R 3-2, 5-10 membered heteroaryl substituted with 1, 2 or 3R 3-1 or 3-6 membered heterocycloalkyl substituted with 1, 2 or 3R 3-4;
R 3-1 is C 1-C6 alkyl; r 3-2 is independently halogen, C 1-C6 alkyl or (c=o) NR 3-2-3R3-2-4;R3-2-3 and R 3-2-4 is independently H or C 1-C6 alkyl;
R 3-3 is independently H or C 1-C6 alkyl;
R 3-4 is independently C 1-C6 alkyl;
II: -in L 1-L2-L3 -a, L 1 is- (CH 2) n-, n is 2,3 or 4, wherein 0,1, 2 or 3-CH 2 -are replaced by CHR L-1 or-CR L-1RL-1-;RL-1 are independently C 1-C6 alkyl;
l 2 is -(C＝O)NR2-1-、-NR2-1(C＝O)-、-NR2-1(C＝O)NR2-1-、-(C＝O)O-、-(C＝O)-、-(C＝O)NR2-1NR2-1(C＝O)-
Or a 5-6 membered heteroarylene group;
R 2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted with 1,2 or 3R 2-1-1; r 2-1-1 is independently OH;
L 3 is a bond;
A is C 6-C10 aryl, 3-6 membered heterocycloalkyl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2 or 3-6 membered heterocycloalkyl substituted with 1,2 or 3R 3-4;
r 3-1 is CN, COOR 3-1-3、C1-C6 alkyl, halogen, CONR 3-1-1R3-1-2、C3-C6 cycloalkyl, 3-6 membered heterocycloalkyl or C 1-C6 alkyl substituted with 1,2 or 3R 3-1-4; r 3-1-1、R3-1-2 and R 3-1-3 are independently H or C 1-C6 alkyl; r 3-1-4 is independently halogen or 5-10 membered heteroaryl;
r 3-2 is independently OH, NR 3-2-1(S＝O)2R3-2-2、NR3-2-3R3-2-4;R3-2-1、R3-2-2、R3-2-3 and R 3-2-4 are independently C 1-C6 alkyl;
r 3-4 is independently C 1-C6 alkyl.
13. A compound of formula (I) or a pharmaceutically acceptable salt thereof, as claimed in claim 1, which meets one or more of the following conditions:
(1) -L 1-L2-L3 -is
Wherein the f side is connected with A;
(2) -L 1-L2-L3 -A is:
14. The compound of any one of claims 1 to 13, wherein the compound of formula (I) is any one of the following:
Scheme 1:
Wherein R 1、L1、L2、L3 and A are as defined in any one of claims 1 to 13;
Scheme 2:
Wherein,
R 1 is-OH or-NR 1-1SO2R1-2;R1-1 and R 1-2 is independently H or C 1-C6 alkyl;
L 2 is-NR L-2-、-C(＝O)NR2-1-、-NR2-1 C (=o) -or-NR 2-1C(＝O)NR2-1-,R2-1 is independently H, C 1-C6 alkyl or C 1-C6 alkyl substituted by 1,2 or 3R 2-1-1; r 2-1-1 is independently C 1-C6 alkoxy; r L-2 is H or C 1-C6 alkyl;
L 3 is a bond or- (CH 2) m-, m is 1, 5, 6, 7 or 8, 0 or 1 of the- (CH 2) m-s-CH 2 -s are replaced by-ch=ch-, -NH-, or-O-;
A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1, C 6-C10 aryl substituted with 1,2 or 3R 3-2;
R 3-1 and R 3-2 are as defined in any one of claims 1 to 13;
Scheme 3:
Wherein, Is 3-6 membered heterocycloalkylene; q is (CH 2)2-4; A is C 6-C10 aryl, 5-10 membered heteroaryl substituted with 1,2 or 3R 3-1 or C 6-C10 aryl substituted with 1,2 or 3R 3-2; R 3-1 and R 3-2 are as defined in any one of claims 1 to 13;
Scheme 4:
Wherein L 2 is-NR 2-1C(＝O)N R2-1-、-C(＝O)NR2-1 -or-N R 2-1-C(＝O)-,R2-1 is H or C 1-C6 alkyl;
L 3 is- (CH 2) m-and m is 9,10 or 11;
A is H, CN, -NR 3-3R3-3 OR-OR 3-3;
R 3-3 is independently H or C 1-C6 alkyl.
15. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula (I) is any one of the following compounds:
Preferably, the pharmaceutically acceptable salts of the compounds shown in the formula (I) are:
16. A pharmaceutical composition comprising a substance X which is a compound of formula (I) as defined in any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
17. Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1 to 15, or a pharmaceutical composition as claimed in claim 16, in the preparation of an SMO inhibitor.
18. Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, as claimed in any one of claims 1 to 15, or a pharmaceutical composition as claimed in claim 16, for the manufacture of a medicament for the treatment and/or prophylaxis of SMO-related disorders.
19. Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16 according to any one of claims 1-15 in the manufacture of a medicament for the treatment and/or prevention of a disease associated with aberrant activation of Hh signaling pathway.
20. Use of a compound of formula (I), a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for the manufacture of a medicament for the treatment and/or prophylaxis of basal cell carcinoma or medulloblastoma.
CN202311773333.3A 2022-12-27 2023-12-21 Tricyclic compound, pharmaceutical composition and application thereof Pending CN118255685A (en)

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CN2023101711796 2023-02-27

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