WO2023001133A1 - Prmt5抑制剂 - Google Patents

Prmt5抑制剂 Download PDF

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Publication number
WO2023001133A1
WO2023001133A1 PCT/CN2022/106401 CN2022106401W WO2023001133A1 WO 2023001133 A1 WO2023001133 A1 WO 2023001133A1 CN 2022106401 W CN2022106401 W CN 2022106401W WO 2023001133 A1 WO2023001133 A1 WO 2023001133A1
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Prior art keywords
mmol
compound
amino
reaction solution
solution
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PCT/CN2022/106401
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English (en)
French (fr)
Inventor
付家胜
覃华
朱伟波
石谷沁
孙大庆
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上海齐鲁制药研究中心有限公司
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Priority to CN202280027588.2A priority Critical patent/CN117157297A/zh
Publication of WO2023001133A1 publication Critical patent/WO2023001133A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems

Definitions

  • the invention belongs to the field of medicinal chemistry, and specifically relates to a quinoline-substituted PRMT5 inhibitor, a pharmaceutical composition containing the compound and a method for using the compound of the invention to treat cell proliferation diseases, such as cancer.
  • Protein arginine methyltransferase plays an important role in the methylation of proteins, such as participating in variable splicing, post-transcriptional regulation, RNA processing, cell proliferation, cell differentiation, cell Apoptosis and tumor formation etc.
  • PRMT1-11 protein arginine methyltransferase
  • PRMTs can be divided into three types: type I includes PRMT1, PRMT2, PRMT3, PRMT4, PRMT6 And PRMT8, the catalyzed forms are monomethyl (MMA) and asymmetric dimethyl (aDMA); type II is symmetric dimethyl (sDMA), including PRMT5 and PRMT9; type III is PRMT7.
  • type I includes PRMT1, PRMT2, PRMT3, PRMT4, PRMT6 And PRMT8, the catalyzed forms are monomethyl (MMA) and asymmetric dimethyl (aDMA); type II is symmetric dimethyl (sDMA), including PRMT5 and PRMT9; type III is PRMT7.
  • type I includes PRMT1, PRMT2, PRMT3, PRMT4, PRMT6 And PRMT8, the catalyzed forms are monomethyl (MMA) and asymmetric dimethyl (aDMA); type II is symmetric dimethyl (sDMA), including PRMT5 and PRMT9; type III is PRMT7.
  • MMA monomethyl
  • aDMA asymmetric dimethyl
  • PRMT5 was isolated for the first time in a protein complex interacting with Janus tyrosine kinase 2 (Jak2), so it is also called Jak-binding protein 1 (JBP1).
  • JBP1 Jak-binding protein 1
  • PRMT5 can symmetrically methylate arginine residues of histone or non-histone substrates, affect multiple target genes and multiple signaling pathways, and thus play a variety of biological roles.
  • PRMT5 is also an oncogene, which is highly expressed in a variety of tumors, and its expression level is closely related to the occurrence, development and prognosis of tumors.
  • CYP450 enzymes The metabolic pathway in which CYP450 enzymes participate is a very important metabolic pathway in the process of compound clearance in vivo. Inhibition of the activity of this family of enzymes can bring about changes in drug clearance and pharmacokinetics in vivo, and drug interactions mediated by CYP450 enzymes ( Drug-drug interaction (DDI) is an important factor in the consideration of drug efficacy and drug safety (J Pharmacol Exp Ther.2006 Jan[J].2006,316(1):336-48).
  • CYP3A4 is the main metabolic enzyme of the CYP450 family, and inhibition of DDI brought about by CYP3A4 will lead to serious safety issues (The AAPS Journal volume 24, Article number: 12(2022)).
  • CYP3A4 is the most abundant P450 isoenzyme in the liver and intestinal wall, and can participate in the metabolism of about 50% of clinical drugs.
  • drugs that inhibit or induce CYP3A4 can affect the pharmacokinetics of other drugs combined.
  • Kinetics, thereby affecting drug exposure in plasma leads to pharmacokinetic DDI.
  • Inhibitory (including reversible, mechanism inactivation) drug interactions often improve drug efficacy, but drugs with a narrow therapeutic window are likely to cause clinical adverse reactions, which can be life-threatening in severe cases.
  • the pharmacokinetic DDI induced by induction seldom causes drug safety problems, but it can reduce the efficacy of the drug.
  • predicting the DDI that may be caused by new drugs is an important part of evaluating the properties of candidate drugs in the development of new drugs (Zhang Qinghao et al.: Quantitative prediction of in vivo drug-drug interactions based on in vitro CYP3A4 inhibition and induction data. Acta Pharmaceutica Sinica[J].2010, 45(8):952-959).
  • Inhibition of P450 enzymes by drugs is usually divided into reversible inhibition (including competitive inhibition, non-competitive inhibition and anti-competitive inhibition) and irreversible inhibition.
  • Reversible inhibitors form complexes with enzymes (or enzyme-substrate complexes) through non-covalent bonds, which hinder the normal progress of enzymatic reactions between enzymes and substrates. After removing inhibitors, enzyme activity is not affected. It can continue to carry out normal enzymatic reaction with the substrate.
  • irreversible inhibition the inhibitory effect on the enzyme does not disappear immediately after removal of the inhibitor, but exhibits a time-dependent property. This phenomenon is generally called time-dependent inhibition (time-dependent inhibition, TDI).
  • MBI mechanism-based inhibition
  • TDI Compared with reversible inhibition, TDI will bring more serious drug safety problems, because many drugs that need to be used in combination also require long-term medication, which will inevitably lead to long-term inhibition of the inhibited CYP subtypes, even if the drug that produces TDI is stopped. Drugs, their inhibitory effects will continue for a period of time; at the same time, because the inhibitors that produce MBI are also the substrates of P450 enzymes, when the enzyme activity is inhibited, its own metabolism will also be hindered, resulting in non-stable exposure in the body In addition, the covalent modification of P450 enzymes by reactive metabolites may lead to the production of haptens, which may cause autoimmune reactions with serious consequences (Xie Shanshan et al. Time-dependent inhibition of cytochrome P450 enzymes and its role in The role of new drug development [J]. Chinese Journal of New Drugs and Clinical. Chin J New Drugs Clin Rem, 2013,32(6):419-424).
  • the present invention provides a compound represented by formula (I') or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H or NR 1a R 1b ;
  • R 1a and R 1b are each independently selected from H or C 1-4 alkyl; said C 1-4 alkyl may be optionally substituted by one or more R 1c groups, said R 1c groups are C 3-6 cycloalkyl;
  • Y is selected from N or CR 2 ;
  • R 2 is selected from H, halogen, C 1-4 alkyl substituted by halogen, CN or -SO 2 -C 1-4 alkyl;
  • X is selected from O, S, NH or CH2 ;
  • Ring A is selected from C 5-6 cycloalkyl, C 5-6 cycloalkenyl or 5-10 membered heterocyclyl;
  • R 3 is selected from hydroxyl or C 1-4 alkyl
  • n 0, 1, 2 or 3;
  • n is selected from 0 or 1;
  • Ring B selected from
  • R 4 is selected from NR 4a R 4b or C 1-4 alkyl
  • R 4a and R 4b are each independently selected from H or C 1-4 alkyl
  • R 5 and R 6 are each independently selected from H, deuterium, halogen or C 1-4 alkyl;
  • R 7 and R 8 are each independently H;
  • R 5 and R 8 when When it is a double bond, R 5 and R 8 do not exist, and R 6 and R 7 form a phenyl group or a C 5-6 cycloalkyl group together with the C atom connected to it.
  • the present invention provides a compound represented by formula (I') or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula (I') or a pharmaceutically acceptable salt thereof is the formula ( I) the compound shown in or a pharmaceutically acceptable salt thereof,
  • R 1 is selected from H or NR 1a R 1b ;
  • R 1a and R 1b are each independently selected from H or C 1-4 alkyl; said C 1-4 alkyl can be optionally substituted by one or more R 1c groups, and R 1c groups are C 3 -6 cycloalkyl;
  • R 2 is selected from H, halogen, C 1-4 alkyl substituted by halogen, CN or -SO 2 -C 1-4 alkyl;
  • R 3 is selected from hydroxyl or C 1-4 alkyl
  • R 4 is selected from hydroxyl, NR 4a R 4b or C 1-4 alkyl, preferably, R 4 is selected from NR 4a R 4b or C 1-4 alkyl;
  • R 4a and R 4b are each independently selected from H or C 1-4 alkyl
  • X is selected from O, S, NH or CH2 ;
  • Ring A is selected from C 5-6 cycloalkyl, C 5-6 cycloalkenyl or 5-10 membered heterocyclyl;
  • n 0, 1, 2 or 3;
  • n is selected from 0 or 1;
  • R 5 and R 6 are each independently selected from H, deuterium, halogen or C 1-4 alkyl;
  • R 7 and R 8 are each independently H;
  • R 5 and R 8 when When it is a double bond, R 5 and R 8 do not exist, and R 6 and R 7 form a phenyl group or a C 5-6 cycloalkyl group together with the C atom connected to it.
  • R 1 is selected from H or NR 1a R 1b ;
  • R 1a and R 1b are each independently selected from H or C 1-4 alkyl; said C 1-4 alkyl can be optionally substituted by one or more R 1c groups, and R 1c groups are C 3 -6 cycloalkyl;
  • R is selected from H or halogen
  • R 3 is selected from hydroxyl or C 1-4 alkyl
  • R 4 is selected from hydroxyl, NR 4a R 4b or C 1-4 alkyl, preferably, R 4 is selected from NR 4a R 4b or C 1-4 alkyl;
  • R 4a and R 4b are each independently selected from H or C 1-4 alkyl
  • X is selected from O, S, NH or CH2 ;
  • Ring A is selected from C 5-6 cycloalkyl, C 5-6 cycloalkenyl or 5-10 membered heterocyclyl;
  • n 0, 1, 2 or 3;
  • n is selected from 0 or 1;
  • R 5 and R 6 are each independently selected from H, deuterium or halogen;
  • R 7 and R 8 are each independently H;
  • R 5 and R 8 when When it is a double bond, R 5 and R 8 do not exist, and R 6 and R 7 form a phenyl group or a C 5-6 cycloalkyl group together with the C atom connected to it.
  • said R 1 is selected from H, amino or
  • said R 2 is selected from H, F, Cl or Br.
  • the R 2 is selected from halogen-substituted C 1-4 alkyl, CN or -SO 2 -C 1-4 alkyl.
  • the R 2 is selected from CF 3 , CN or -SO 2 -CH 3 .
  • the L 1 is selected from -CH ⁇ CH-CH 2 -.
  • the ring A is selected from C5-6 cycloalkyl, C5-6 cycloalkenyl or 5-10 membered heterocyclic group, and the heterocyclic group contains 1 or more A heteroatom selected from N or O.
  • ring A is selected from C5-6 cycloalkyl, C5-6 cycloalkenyl, 5-10 membered monoheterocycle, 5-10 membered spiroheterocycle, 5-10 membered bridged heterocycle Or 5-10 membered condensed heterocycle.
  • ring A is selected from C5-6 cycloalkyl, C5-6 cycloalkenyl, 5-6 membered monoheterocycle, 9-10 membered spiroheterocycle, 7-10 membered bridged heterocycle Or 8-10 membered condensed heterocycle.
  • the ring A is selected from
  • the R 3 is selected from hydroxyl or methyl; m is selected from 0, 1, 2 or 3.
  • the R 4 is selected from amino, methyl or -NHCH 3 .
  • the represents a single or double bond
  • R 5 and R 6 are each independently selected from H, deuterium, F or methyl;
  • R 7 and R 8 are each independently H;
  • R 5 and R 8 do not exist, and R 6 and R 7 form a phenyl group or a cyclopentyl group together with the C atom connected to it.
  • the compound or a pharmaceutically acceptable salt thereof, wherein the compound has the structure shown in the following formula (II):
  • R 9 is selected from H or C 1-4 alkyl; preferably, R 9 is selected from H or methyl;
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , L 1 and m are as defined in formula (I′) or formula (I).
  • R 9 is selected from H or C 1-4 alkyl; preferably, R 9 is selected from H or methyl; m is selected from 1 or 2; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 and L 1 are as defined in formula (I′) or formula (I).
  • the above-mentioned compound, or a pharmaceutically acceptable salt thereof is selected from:
  • R 1 , R 2 , R 4 , R 5 , R 6 , R 7 , R 8 and R 9 are as defined in formula (II).
  • the above compound, its isomer or pharmaceutically acceptable salt is selected from:
  • R 1 , R 2 , R 4 and R 9 are as defined in formula (II).
  • the present invention also provides the following compounds, their isomers or pharmaceutically acceptable salts, which are selected from:
  • the present invention also relates to a pharmaceutical composition, which comprises the compound, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient.
  • the present invention also relates to the application of the compound or its pharmaceutically acceptable salt or its pharmaceutical composition in preparing medicine for treating cancer.
  • the present invention also relates to the use of the compound or its pharmaceutically acceptable salt, or its pharmaceutical composition in treating cancer.
  • the present invention also relates to a method for treating cancer, which comprises administering an effective dose of the compound or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a subject in need.
  • the compound of the present invention has good inhibitory effect on PRMT5 methylase and human pancreatic cancer cell line MIA PaCa-2, and has the potential advantage of higher heart safety at the same time, has little inhibitory effect on CYP3A4 enzyme, and drug interaction occurs ( DDI) is less likely, has a lower risk of significant time-dependent inhibition (TDI), and has favorable pharmacokinetic properties. Compared with the prior art, the compound of the invention has higher safety.
  • pharmaceutically acceptable means, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction or other problems or complications, and with a reasonable benefit/risk ratio Comparable to those compounds, materials, compositions and/or dosage forms.
  • pharmaceutically acceptable salt refers to derivatives prepared from the compounds of the present invention with relatively non-toxic acids or bases. These salts can be prepared during compound synthesis, isolation, purification, or alone by reacting the free form of the purified compound with an appropriate acid or base.
  • the compound contains relatively acidic functional groups, it reacts with alkali metal, alkaline earth metal hydroxides or organic amines to obtain base addition salts, including cations based on alkali metals and alkaline earth metals.
  • the compound contains a relatively basic functional group, it reacts with an organic acid or an inorganic acid to form an acid addition salt.
  • the compounds of the present invention exist as isomers, such as cis-trans isomers, enantiomers, diastereomers, and racemic and other mixtures thereof, all of which are within the scope of the present invention .
  • enantiomer refers to stereoisomers that are mirror images of each other.
  • diastereomer refers to stereoisomers whose molecules have two or more chiral centers and which are in a non-mirror-image relationship.
  • cis-trans isomer refers to the configuration in which the double bond or the single bond of the ring carbon atom in the molecule cannot freely rotate.
  • keys with wedge-shaped solid lines and dotted wedge keys Indicates the absolute configuration of a stereocenter.
  • Stereoisomers of the compounds of the present invention may be prepared by chiral synthesis or chiral reagents or other conventional techniques.
  • one enantiomer of a certain compound of the present invention can be prepared by asymmetric catalytic technology or chiral auxiliary derivatization technology.
  • a compound with a single stereo configuration can be obtained from a mixture by chiral resolution technology.
  • it can be directly prepared by using chiral starting materials.
  • the separation of optically pure compounds in the present invention is usually accomplished by using preparative chromatography, and a chiral chromatographic column is used to achieve the purpose of separating chiral compounds.
  • chiral carbon atoms are marked with *.
  • the carbon atoms on the cyclopentyl group are chiral carbons, including Compound of two structures.
  • the isomers separated by chiral chromatographic column in the present invention are represented by P1 and P2, for example, Example 6-P1 and Example 6-P2 represent the two isomers obtained by chiral resolution of the compound in Example 6.
  • the present invention also includes isotopically labeled compounds, including isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, respectively , 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl.
  • isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, respectively , 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl.
  • Compounds of the present invention that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention.
  • the isotope is selected from the group consisting of 2 H, 3 H, 11 C and 18 F. More preferably, the radioisotope is2H .
  • deuterated compounds are intended to be encompassed
  • substituent R3 When a substituent is bonded across a ring, the substituent may be bonded to any atom on the ring.
  • the structural unit Indicates that substituent R3 can be substituted at any position on ring A.
  • pharmaceutically acceptable carrier refers to a medium generally acceptable in the art for delivering biologically active agents to animals, especially mammals, including adjuvants, excipients or Excipients such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, Lubricants and dispersants.
  • Pharmaceutically acceptable carriers are formulated according to a number of factors that are within the purview of those of ordinary skill in the art.
  • compositions containing the agent include, but are not limited to: the type and nature of the active agent being formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the intended therapeutic indication.
  • Pharmaceutically acceptable carriers include both aqueous and non-aqueous media and various solid and semisolid dosage forms. Such carriers include many different ingredients and additives in addition to the active agent, and the inclusion of additional ingredients in formulations for a variety of reasons (eg, stabilizing the active agent, binders, etc.) is well known to those of ordinary skill in the art.
  • excipient generally refers to a carrier, diluent and/or medium required to formulate an effective pharmaceutical composition.
  • prophylactically or therapeutically effective amount means that a compound of the present invention or a pharmaceutically acceptable salt thereof means a sufficient amount of the compound to treat the disorder with a reasonable effect/risk ratio applicable to any medical treatment and/or prevention.
  • total daily dosage of the compound represented by formula (I) of the present invention or its pharmaceutically acceptable salts and compositions must be determined by the attending physician within the scope of reliable medical judgment.
  • the particular therapeutically effective dosage level will depend on a number of factors, including the disorder being treated and the severity of the disorder; the activity of the particular compound employed; the particular composition employed; The age, weight, general health, sex and diet of the patient; the timing, route of administration, and rate of excretion of the specific compound employed; the duration of treatment; drugs used in combination or concomitantly with the specific compound employed; and Similar factors are well known in the medical arts. For example, it is practice in the art to start doses of the compound at levels lower than that required to obtain the desired therapeutic effect and to gradually increase the dosage until the desired effect is obtained.
  • any variable eg R 1c
  • R 1c any variable (eg R 1c ) occurs more than once in the composition or structure of a compound, its definition is independent at each occurrence. For example, if a group is substituted with 0-2 R 1c , said group may optionally be substituted with up to two R 1c , and there are independent options for each occurrence of R b .
  • cyclic refers to saturated, partially saturated or unsaturated monocyclic as well as polycyclic rings.
  • heterocyclyl refers to a substituted or unsubstituted saturated or unsaturated non-aromatic ring containing 1-3 heteroatoms selected from N, O or S.
  • the "heterocyclic group” mentioned in the present invention refers to a non-aromatic cyclic group derived from the removal of a hydrogen atom containing at least one heteroatom as a ring atom; including saturated or partially saturated monocyclic heterocyclic groups and Polycyclic heterocyclyl; said heterocyclyl is independent of the position of attachment (ie, bonding may be through a carbon atom or a heteroatom).
  • the polycyclic heterocyclic group includes a fused heterocyclic group, a spiro heterocyclic group, and a bridged heterocyclic group.
  • the "fused heterocyclic group” refers to a ring structure formed by two or more rings sharing two adjacent ring atoms, and at least one of the rings is a heterocyclic ring; the fused heterocyclic ring
  • the group includes a ring structure formed by condensing a monoheterocyclic group and a monoheterocyclic group or a cycloalkyl group or an aryl group or a heteroaryl group, and also includes a fused ring structure formed by a heteroaryl group and a cycloalkyl group or a monoheterocyclic group ring structure.
  • the "spiroheterocyclic group” refers to a ring structure formed by two or more rings sharing one ring atom with each other, and at least one ring is a heterocycle.
  • bridged heterocyclic group refers to a ring structure formed by two or more rings sharing non-adjacent ring atoms with each other, and at least one ring is a heterocyclic ring.
  • the heterocyclic group described in the present invention is preferably a 5-10 membered monoheterocyclic group, a 5-10 membered condensed heterocyclic group, a 5-10 membered spiroheterocyclic group or a 5-10 membered bridged heterocyclic group; further, the heterocyclic group
  • the ring group is preferably a 5-6 membered monoheterocyclic group, an 8-10 membered condensed heterocyclic group, an 8-10 membered spiroheterocyclic group or a 7-9 membered bridged heterocyclic group.
  • 5-10 membered heterocyclyl include, but are not limited to, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, 1,3- Dioxolane, 1,4-dioxane, Wait.
  • heterocyclic group when both ends of the heterocyclic group are substituted, it is a heterocyclic group, such as a structural unit
  • ring A when ring A is selected from 5-10 membered heterocyclyl, it essentially means that ring A is 5-10 membered heterocyclylene, including but not limited to
  • aryl refers to an unsaturated, usually aromatic, hydrocarbon group which may be a single ring or multiple rings fused together. Examples of aryl include, but are not limited to, phenyl, naphthyl.
  • heteroaryl means a stable monocyclic or polycyclic aromatic hydrocarbon, preferably containing carbon atoms and 1, 2, 3 or 4 ring heterocyclic rings independently selected from N, O and S atom.
  • heteroaryl groups include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, pyrazinyl, isoxazolyl, thiazolyl, furyl, thienyl, pyrimidyl, pyridyl,
  • heteroaryl group when both ends of the heteroaryl group are substituted, it is a heteroarylene group, for example, when L in the compound of formula (I') or formula ( I ) is selected from 5-6 membered heteroaryl groups, it essentially represents L 1 is a 5-6 membered heteroarylene group, including but not limited to
  • cycloalkyl means a saturated monocyclic or polycyclic hydrocarbon group.
  • Cycloalkyl is preferably C 3-8 cycloalkyl, more preferably C 3-6 cycloalkyl, further preferably C 5-6 cycloalkyl, examples of cycloalkyl include but not limited to cyclopropyl, cyclobutyl base, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl or
  • cycloalkylene group such as a structural unit
  • ring A is selected from C 5-6 cycloalkyl, it essentially means that ring A is C 5-6 cycloalkylene, including but not limited to
  • cycloalkenyl means a monocyclic or polycyclic hydrocarbon group containing a double bond.
  • Cycloalkenyl is preferably C 3-6 cycloalkenyl, more preferably C 5-6 cycloalkenyl, examples include but not limited to cyclopropene, cyclobutene, cyclopentene, cyclohexene, 1,3-cyclohexanedi ene or 1,4-cyclohexadiene.
  • cycloalkenylene group such as a structural unit
  • ring A is selected from C 5-6 cycloalkenyl, it essentially represents that ring A is C 5-6 cycloalkenylene, including but not limited to
  • Ring B in the present invention is selected from situation, when When it is a double bond, R 5 and R 8 do not exist, R 6 and R 7 form a phenyl group or a C 5-6 cycloalkyl group together with the C atom connected to it, and the corresponding Refers to
  • alkyl is used to denote a straight or branched chain saturated hydrocarbon group. It is preferably a C 1-6 alkyl group, more preferably a C 1-4 alkyl group. Examples of the alkyl group include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl, n-hexyl, etc.
  • halogen means a fluorine, chlorine, bromine or iodine atom.
  • the name of the title compound was converted from the compound structure by means of Chemdraw. If there is any inconsistency between the name of the compound and the structure of the compound, it can be determined by comprehensively related information and reaction routes; if it cannot be confirmed by other methods, the structural formula of the given compound shall prevail.
  • the preparation methods of some compounds in the present invention refer to the preparation methods of the aforementioned similar compounds. Those skilled in the art should know that when using or referring to the preparation methods cited therein, the feed ratio of reactants, reaction solvent, reaction temperature, etc. can be adjusted appropriately according to different reactants.
  • the compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and the methods well known to those skilled in the art Equivalent alternatives, preferred embodiments include but are not limited to the examples of the present invention.
  • the structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass chromatography (LC-MS), or ultra-high performance liquid chromatography-mass chromatography (UPLC-MS). NMR chemical shifts ( ⁇ ) are given in parts per million (ppm).
  • the determination of NMR is to use Bruker Neo 400M or Bruker Ascend 400 nuclear magnetic instrument, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ), heavy water (D 2 O), internal standard is tetramethylsilane (TMS).
  • Agilent 1260-6125B single quadrupole mass spectrometer was used for liquid chromatography-mass chromatography LC-MS, and the column was Welch Biomate column (C18, 2.7 ⁇ m, 4.6 ⁇ 50mm) or waters H-Class SQD2, and the column was Welch Ultimate column (XB- C18, 1.8 ⁇ m, 2.1 ⁇ 50mm), the ion source of the mass spectrometer is electrospray ionization.
  • HPLC uses Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high performance liquid chromatography.
  • Preparative HPLC uses Waters 2555-2489 (10 ⁇ m, ODS 250cm ⁇ 5cm) or GILSON Trilution LC, and the column is Welch XB-C18 column (5 ⁇ m, 21.2 ⁇ 150mm).
  • Chiral HPLC uses waters acquisition UPC2; the column is Daicel chiralpak AD-H (5 ⁇ m, 4.6 ⁇ 250mm), Daicel chiralpak OD-H (5 ⁇ m, 4.6 ⁇ 250mm), Daicel chiralpak IG-3 (3 ⁇ m, 4.6 ⁇ 150mm), Chiral Technologies Europe AD-3 (3 ⁇ m, 3.0 ⁇ 150mm) and Trefoil TM Technology Trefoil TM AMY1 (2.5 ⁇ m, 3.0 ⁇ 150mm).
  • Supercritical fluid chromatography uses waters SFC 80Q, and the column is Daicel Chiralcel OD/OJ/OZ (20 ⁇ 250mm, 10 ⁇ m) or Daicel Chiralpak IC/IG/IH/AD/AS (20 ⁇ 250mm, 10 ⁇ m).
  • Thin-layer chromatography silica gel plates use Yantai Jiangyou Silica Gel Development Co., Ltd. GF254 silica gel plates or Rushan Shangbang New Materials Co., Ltd. GF254 silica gel plates.
  • the specifications used by TLC are 0.15mm to 0.20mm, preparative 20 ⁇ 20cm, column chromatography Generally used in Chenghua Chemical Industry 200-300 mesh silica gel as carrier.
  • the starting materials in the examples of the present invention are known and commercially available, or can be synthesized using or following methods known in the art.
  • Step A Add 7-nitroquinoline (25g, 143mmol) and acetic acid solution (50mL) to the reaction flask.
  • the replacement reaction system is a nitrogen system, and then cooled to 0°C.
  • N-bromosuccinimide (50.6 g, 284 mmol) was added dropwise to the reaction system, and the reaction system was stirred at 100° C. for 2 hours.
  • Step C Sulfuric acid (108 mL) was added to the reaction flask, then a solution (86 mL) of 3-bromoquinolin-7-amine (6.2 g, 27.8 mmol) was added slowly at 0°C. After stirring at 0°C for 10 minutes, a solution (6 mL) of sodium nitrite (3.8 g, 55.1 mmol) was slowly added dropwise. Then the stirring was continued for 30 minutes, and a solution of sodium iodide (12.5 g, 83.4 mmol) was slowly added to the reaction system at 0°C. The reaction solution was stirred at 60°C for 2 hours.
  • Step D 3-Bromo-7-iodoquinoline (500 mg, 1.5 mmol) was dissolved in dichloromethane (10 mL) at room temperature. m-chloroperbenzoic acid (1.03 g, 6 mmol) was slowly added at 0° C., and the reaction solution was stirred at room temperature for 12 hours.
  • the reaction was monitored by LCMS, quenched by adding water (30 mL) to the reaction solution at 0°C, then slowly adding a saturated aqueous solution of ammonium chloride to adjust the pH to 7, and concentrating under reduced pressure.
  • the mixture was extracted with ethyl acetate (15 mL ⁇ 3 times), and the organic phases were combined.
  • the organic phase was washed with saturated brine (50 mL), then dried over anhydrous sodium sulfate, filtered, and the collected filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography to obtain 3-bromo-7-iodoquinoline 1-oxide (160 mg).
  • Step E 3-Bromo-7-iodoquinoline 1-oxide (160 mg, 0.45 mmol) was dissolved in chloroform (10 mL) at room temperature. Then, phosphorus oxychloride (10 mL) was slowly added at 0° C., and the reaction system was evacuated and replaced with nitrogen several times. The reaction solution was stirred for 12 hours in an oil bath at 70°C.
  • Step A 6-hydrazinopyrimidin-4-ol (2.0 g, 15.9 mmol) and cyclopentanone (1.3 g, 15.5 mmol) were dissolved in ethanol (15 mL). The reaction system was heated to 80°C and stirred for 3 hours.
  • Step B Dissolve 5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-ol (2.7 g, 15.4 mmol) in trichloroxy Phosphorus (20 mL). The reaction system was heated to 100°C and stirred for 3 hours. After LCMS monitoring showed disappearance of the starting material, most of the phosphorus oxychloride was removed under vacuum distillation.
  • reaction solution was diluted with ethyl acetate (50mL) and slowly dropped into an aqueous solution of sodium hydroxide (1N, 50mL) to extract the organic phase, which was washed with saturated brine (300mL x 3 times), dried over anhydrous sodium sulfate, and filtered , concentrated under reduced pressure.
  • the resulting mixture was purified by silica gel column chromatography to obtain 4-chloro-5,6,7,8-tetrahydrocyclopenta[4,5]pyrrolo[2,3-d]pyrimidine (370 mg).
  • Step C At room temperature, add 4-chloro-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine (243.0mg, 1.3mmol) Dissolve in tetrahydrofuran (4 mL). Subsequently, potassium tert-butoxide (141.3 mg, 1.3 mmol) was added thereto. The reaction was stirred at room temperature for 2 hours.
  • Step D In a single-necked flask, add trifluoromethanesulfonic anhydride (1.16g, 4.11mmol) in dichloromethane (20mL) solution, add (3aS, 4S, 6R, 6aR)- 2,2-Dimethyl-6-vinyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-ol (500mg, 2.74mmol, reference J.Org. Chem.2004, 69, 3993-3996, the entire content of which is incorporated by reference in this application) and pyridine (649 mg, 8.22 mmol) in dichloromethane (10 mL). The reaction solution was stirred at this temperature for half an hour.
  • Step E At 0°C, potassium 4-chloro-6,7-dihydro-5H-cyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine-8-ylate (291.1 mg , 1.26mmol) was dissolved in N,N-dimethylformamide (4mL), then (3aR,4S,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclo Pentadieno[d][1,3]dioxan-4-yl trifluoromethanesulfonate (331.8 mg, 1.05 mmol) dissolved in N,N-dimethylformamide (0.5 mL) was added slowly in the reaction solution. The reaction was stirred at room temperature for 18 hours.
  • Step F At room temperature, 4-chloro-8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d] [1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine (180mg, 0.50mmol ), 7-bromoquinolin-2-amine (133.5 mg, 0.60 mmol) and tetraethylammonium chloride (91.1 mg, 0.55 mmol) were dissolved in N,N-dimethylformamide (5 mL).
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (22.4 mg, 0.1 mmol), N,N-diisopropylethylamine (322.5 mg, 2.5 mmol) were added under nitrogen atmosphere. The reaction solution was stirred for 18 hours in an oil bath at 80°C.
  • Step G 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-chloro-6,7-dihydrocyclopentadiene[4, 5]pyrrolo[2,3-d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxane- 4-yl)vinyl)quinolin-2-amine (45mg, 0.09mmol) was dissolved in ammonia water/isopropanol (2mL/0.8mL), and the reaction system was sealed and heated to 120°C and stirred for 18 hours.
  • Step H 8-((3aS,4R,6R,6aR)-6-((E)-2-(2-aminoquinolin-7-yl)vinyl)-2,2-dimethyltetrahydro
  • Base-4H cyclopentadieno[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrole[2, 3-d]
  • Pyrimidin-4-amine 15mg, 0.03mmol
  • palladium carbon 5mg
  • Step I At room temperature, 8-((3aS,4R,6S,6aR)-6-(2-(2-aminoquinolin-7-yl)ethyl)-2,2-dimethyltetrahydro -4H-Cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4,5]pyrrolo[2, 3-d] Pyrimidin-4-amine (10 mg, 0.02 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 8-((3aS,4R,6R,6aR)-6-((E)-2-(2-aminoquinolin-7-yl)vinyl)-2,2-dimethyl Tetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo [2,3-d]Pyrimidin-4-amine (14 mg, 0.03 mmol) was dissolved in methanol (1.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (1.5 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 4-chloro-8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d ][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine (400mg, 1.11 mmol) was dissolved in ammonia water/isopropanol (7mL/5mL), the reaction system was sealed and heated to 120°C and stirred for 18 hours.
  • Step B 8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-amine (30mg, 0.09mmol) , 3-Bromo-7-iodoquinolin-2-amine (37.1 mg, 0.11 mmol) and tetraethylammonium chloride (16.0 mg, 0.10 mmol) were dissolved in N,N-dimethylformamide (3 mL) .
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (3.9 mg, 0.02 mmol), N,N-diisopropylethylamine (56.8 mg, 0.44 mmol) were added under nitrogen atmosphere. The reaction solution was stirred in an oil bath at 70°C for 18 hours.
  • Step C At room temperature, 8-((3aS,4R,6R,6aR)-6-((E)-2-(2-amino-3-bromoquinolin-7-yl)ethenyl)-2 ,2-Dimethyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4 ,5] Pyrrolo[2,3-d]pyrimidin-4-amine (40 mg, 0.07 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 4-chloro-8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d ][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine (200mg, 0.56 mmol) was dissolved in methylammonia/isopropanol (1mL/5mL), the reaction system was sealed and heated to 120°C and stirred for 18 hours.
  • Step B 8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-N-methyl-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-amine (100mg, 0.28mmol), 7-bromoquinolin-2-amine (75.2mg, 0.34mmol) and tetraethylammonium chloride (51.0mg, 0.31mmol) were dissolved in N,N-dimethylformamide (5mL )middle.
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (12.6 mg, 0.056 mmol), N,N-diisopropylethylamine (180.6 mg, 1.4 mmol) were added under nitrogen atmosphere. The reaction solution was stirred in an oil bath at 100°C for 18 hours.
  • Step C At room temperature, 8-((3aS,4R,6R,6aR)-6-((E)-2-(2-aminoquinolin-7-yl)vinyl)-2,2-di Methyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-N-methyl-5,6,7,8-tetrahydrocyclopentadieno [4,5]pyrrolo[2,3-d]pyrimidin-4-amine (40 mg, 0.07 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A Add compound 8-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1, 3] Dioxin-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-amine (80mg, 0.235mmol ) in tetrahydrofuran (3 mL), and then the system was replaced with a nitrogen atmosphere. At room temperature, a solution of 9-BBN (2.35 mmol) in tetrahydrofuran (0.5 M, 4.7 mL) was slowly added dropwise to the reaction solution.
  • reaction solution was heated to 65° C. and stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, and then an aqueous solution (1 mL) of potassium phosphate (249.1 mg, 1.175 mmol) was added to the reaction mixture.
  • an aqueous solution (1 mL) of potassium phosphate (249.1 mg, 1.175 mmol) was added to the reaction mixture.
  • a solution of 3-bromo-7-iodoquinolin-2-amine 79 mg, 0.225 mmol
  • PdCl 2 (dppf) 18 mg, 0.024 mmol
  • Step B At room temperature, the compound 8-((3aS,4R,6S,6aR)-6-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-2,2- Dimethyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4,5] Pyrrolo[2,3-d]pyrimidin-4-amine (60 mg, 0.106 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-chloro-6,7-dihydrocyclopentadiene[4,5 ]pyrrolo[2,3-d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxane- 4-yl)vinyl)quinolin-2-amine (250.0mg, 0.50mmol), di-tert-butyl dicarbonate (326.8mg, 1.5mmol) and 4-dimethylaminopyridine (12.2mg, 0.1mmol) were dissolved in in tetrahydrofuran (12 mL). Subsequently, triethylamine (202.4 mg, 2.0 mmol) was added to the above solution. The reaction was stirred at room temperature for 18 hours.
  • Step B Slowly add potassium hydroxide (1.7g, 25mL water) aqueous solution into diethyl ether (60mL) at -10°C, then add 1-methyl-1-nitrosourea (3.0g, 29.1mmol) The reaction was stirred at -10°C for 1 hour. The reaction solution was cooled to -78°C, the aqueous phase was frozen and the organic phase was poured out. The organic phase was first washed with saturated brine (100 mL), then dried with anhydrous sodium sulfate, and filtered to obtain the diazomethane diethyl ether solution prepared now. (50mL).
  • Step C At room temperature, di-tert-butyl (7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-chloro-6,7-dihydrocyclopentadiene Ekeno[4,5]pyrrolo[2,3-d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3 ]Dioxane-4-yl)cyclopropyl)quinolin-2-yl)carbamate (150.0mg, 0.21mmol) was dissolved in ammonia water/isopropanol (10mL/3mL), and the reaction system was sealed Heat to 120°C and stir for 24 hours.
  • Step D At room temperature, 8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-aminoquinolin-7-yl)cyclopropyl)-2,2- Dimethyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4,5 ]pyrrolo[2,3-d]pyrimidin-4-amine (90 mg, 0.19 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Embodiment 6-P1 is a diagrammatic representation of Embodiment 6-P1:
  • HPLC retention time 6.254 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Embodiment 6-P2 is a diagrammatic representation of Embodiment 6-P2:
  • HPLC retention time 6.533 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Step A In a one-necked bottle, add (3aR,6aR)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopentadieno[d][1,3]dioxane -4-Kone (830 mg, 5.39 mmol), benzophenone (196 mg, 1.08 mmol) and methanol (350 mL). Then, nitrogen gas was continuously bubbled for 1 hour, and the resulting reaction solution was stirred for 1 hour under the irradiation of a high-pressure mercury lamp (operated in a dark room).
  • Step B In a one-necked bottle, add (3aR,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-cyclopentadiene[d][1,3 ] Dioxan-4-one (450 mg, 2.42 mmol), tert-butyldiphenylchlorosilane (666 mg, 2.42 mmol) and anhydrous dichloromethane (20 mL). The reaction flask was placed in an ice bath and cooled to 0°C, then imidazole (329 mg, 4.84 mmol) was added in batches. The resulting reaction solution was warmed to room temperature and stirring was continued for 3 hours.
  • Step C Add (3aR,6R,6aR)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H- into a three-neck flask Cyclopenta[d][1,3]dioxan-4-one (640mg, 1.5mmol) in tetrahydrofuran (15mL) solution, and then the system was replaced with a nitrogen atmosphere. The reaction flask was cooled to 0° C. in an ice bath, and then lithium aluminum hydride (3.0 mmol) in tetrahydrofuran (2.5 M, 1.2 mL) was added dropwise. After the dropwise addition, the reaction system continued to stir at 0° C. for 1 hour.
  • Step D In a single-necked bottle, add a solution of trifluoromethanesulfonic anhydride (486mg, 1.73mmol) in dichloromethane (15mL), and add dropwise (3aS, 4S, 6R, 6aR)-6 to the reaction solution under ice-cooling -(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxane- A mixture of 4-alcohol (490 mg, 1.15 mmol) and pyridine (272 mg, 3.45 mmol) in dichloromethane (5 mL). The reaction solution was stirred at this temperature for half an hour.
  • Step E Under ice bath, add 4-chloro-6,7-dihydro-5H-cyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine-8-potassium salt (266mg, 1.15 mmol) of N,N-dimethylformamide (8mL) was added dropwise to (3aR,4S,6R,6aR)-6-(((tert-butyldiphenylsilyl)oxy)methyl) N, N-Dimethylformamide (2 mL) solution. After the dropwise addition, the reaction solution was stirred overnight at room temperature.
  • reaction solution was poured into ice water.
  • the reaction solution was extracted with ethyl acetate (20 mL ⁇ 3 times), the organic phases were combined, washed with saturated brine (10 mL ⁇ 3 times), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • Step F Add 8-((3aS,4R,6R,6aR)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyl in a three-neck flask Tetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-4-chloro-5,6,7,8-tetrahydrocyclopenta[4, 5] tetrahydrofuran (10mL) solution of pyrrolo[2,3-d]pyrimidine (492mg, 0.82mmol), then the system was replaced by a nitrogen atmosphere, and tetrahydrofuran solution of tetrabutylammonium fluoride (2.0mmol) was added dropwise ( 1.0 M, 2.0 mL). After the dropwise addition, the reaction system continued to stir at room temperature for 2 hours.
  • Step G At room temperature, ((3aR,4R,6R,6aS)-6-(4-chloro-6,7-dihydrocyclopentadieno[4,5]pyrrolo[2,3-d ]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)methanol (170mg, 0.47 mmol) and 2-chloroquinolin-7-ol (126 mg, 0.71 mmol) were dissolved in toluene (2 mL).
  • Step H at room temperature, 4-chloro-8-((3aS,4R,6R,6aR)-6-(((2-chloroquinolin-7-yl)oxy)methyl)-2,2 -Dimethyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4, 5] Pyrrolo[2,3-d]pyrimidine (190mg, 0.36mmol) was dissolved in ammonia/isopropanol (5mL/1.5mL), and the reaction system was sealed and heated to 120°C and stirred for 24 hours.
  • ammonia/isopropanol 5mL/1.5mL
  • Step I At room temperature, 8-((3aS,4R,6R,6aR)-6-(((2-aminoquinolin-7-yl)oxy)methyl)-2,2-dimethyl Tetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrole[ 2,3-d]pyrimidin-4-amine (58 mg, 0.12 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • the preparation method refers to Example 7, and finally the target product (1R,2S,3R,5R)-3-(4-amino-6,7-dihydrocyclopentadieno[4,5]pyrrolo[2,3 -d]pyrimidin-8(5H)-yl)-5-(((2-((cyclopropylmethyl)amino)quinolin-7-yl)oxy)methyl)cyclopentane-1,2 -diol.
  • Step A At room temperature, 4-chloro-8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d] [1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidine (137.2 mg, 0.38 mmol ), 3-bromo-7-iodoquinolin-2-amine (140.0mg, 0.40mmol) and tetraethylammonium chloride (69.2mg, 0.42mmol) were dissolved in N,N-dimethylformamide (6mL) middle.
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (17.1 mg, 0.07 mmol), N,N-diisopropylethylamine (245.1 mg, 1.9 mmol) were added under nitrogen atmosphere. The reaction solution was stirred in an oil bath at 70°C for 18 hours.
  • Step B At room temperature, 3-bromo-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-chloro-6,7-dihydrocyclopentadiene [4,5]pyrrolo[2,3-d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]di Oxa-4-yl)vinyl)quinolin-2-amine (145.0mg, 0.25mmol), di-tert-butyl dicarbonate (191.3mg, 0.88mmol) and 4-dimethylaminopyridine (6.1mg, 0.05mmol ) was dissolved in tetrahydrofuran (7 mL). Subsequently, triethylamine (126.5 mg, 1.25 mmol) was added to the above solution. The reaction was stirred at room temperature for 18 hours.
  • Step C Slowly add potassium hydroxide (1.7g) in water (25mL) to diethyl ether (60mL) at -10°C, then add 1-methyl-1-nitrosourea (3.0g, 29.1mmol ) The reaction solution was stirred at -10°C for 1 hour. The reaction solution was cooled to -78°C, the aqueous phase was frozen and the organic phase was poured out. The organic phase was first washed with saturated brine (100 mL), then dried with anhydrous sodium sulfate, and filtered to obtain the diazomethane diethyl ether solution prepared now. (50mL).
  • Step E At room temperature, 8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-amino-3-bromoquinolin-7-yl)cyclopropyl)- 2,2-Dimethyltetrahydro-4H-cyclopentadien[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[ 4,5]pyrrolo[2,3-d]pyrimidin-4-amine (45 mg, 0.08 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • HPLC retention time 7.144 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • HPLC retention time 7.376 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Step A Dissolve 3-bromo-2-chloro-7-iodoquinoline (150 mg, 0.43 mmol) and cyclopropylmethylamine (2 mL) in ethanol (2 mL) at room temperature, and seal the reaction system for heating to 120°C and stirred for 18 hours.
  • Step B 8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-amine (20.0mg, 0.059 mmol), 3-bromo-N-(cyclopropylmethyl)-7-iodoquinolin-2-amine (24.7mg, 0.062mmol) and tetraethylammonium chloride (10.7mg, 0.065mmol) were dissolved in N , in N-dimethylformamide (1.5 mL).
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (2.6 mg, 0.012 mmol), N,N-diisopropylethylamine (38.1 mg, 0.29 mmol) were added under nitrogen atmosphere. The reaction solution was stirred in an oil bath at 70°C for 18 hours.
  • Step C At room temperature, 8-((3aS,4R,6R,6aR)-6-((E)-2-(3-bromo-2-((cyclopropylmethyl)amino)quinoline- 7-yl)vinyl)-2,2-dimethyltetrahydro-4H-cyclopentadien[d][1,3]dioxan-4-yl)-5,6,7,8 - Tetrahydrocyclopenta[4,5]pyrrolo[2,3-d]pyrimidin-4-amine (25 mg, 0.04 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A 8-((3aS,4R,6R,6aR)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyl Tetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-4-chloro-5,6,7,8-tetrahydrocyclopenta[4, 5] Pyrrolo[2,3-d]pyrimidine (405.0mg, 0.67mmol) was dissolved in ammonia water/isopropanol (8mL/3mL), and the reaction system was sealed and heated to 120°C and stirred for 40 hours.
  • Step B At room temperature, 8-((3aS,4R,6R,6aR)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetra Hydrogen-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[ 2,3-d]pyrimidin-4-amine (170.0 mg, 0.29 mmol) was dissolved in tetrahydrofuran (5 mL). Tetrabutylammonium fluoride (1.0M, 0.58 mL) was slowly added dropwise to the reaction system, and the reaction solution was stirred at room temperature for 2 hours.
  • Tetrabutylammonium fluoride 1.0M, 0.58 mL
  • Step C Add ((3aR,4R,6R,6aS)-6-(4-amino-6,7-dihydrocyclopentadieno[4,5]pyrrolo[2,3 -d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)methanol (88mg , 0.26mmol) was dissolved in pyridine (4mL), under the protection of nitrogen, trimethylchlorosilane (86.1mg, 0.79mmol) was slowly added dropwise to the reaction system, and the reaction solution was stirred at 0°C for 3 hours. Benzoyl chloride (43.8mg, 0.31mmol) was slowly added dropwise to the reaction system, and slowly returned to room temperature, and the reaction solution was stirred at room temperature for 18 hours.
  • Step D At room temperature, N-(8-((3aS,4R,6R,6aR)-6-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-cyclopentadiene [d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4,5]pyrrolo[2,3-d]pyrimidine-4 -yl)benzamide (50mg, 0.11mmol) was dissolved in dichloromethane (3mL), and under nitrogen protection, Dess-Martin oxidant (56.8mg, 0.13mmol) was slowly added dropwise to the reaction system, and the reaction solution Stir at room temperature for 2 hours.
  • Step E N-(8-((3aS,4R,6S,6aR)-6-formyl-2,2-dimethyltetrahydro-4H-cyclopentadiene[d][ 1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopentadieno[4,5]pyrrolo[2,3-d]pyrimidin-4-yl)benzene
  • Formamide 49.9 mg, 0.11 mmol
  • 2-chloroquinolin-7-amine 29.9 mg, 0.17 mmol
  • acetic acid 33.6 mg, 0.56 mmol
  • potassium acetate 32.9 mg, 0.34 mmol
  • Step F At room temperature, N-(8-((3aS,4R,6R,6aR)-6-((2-chloroquinolin-7-yl)amino)methyl)-2,2-dimethyl Tetrahydro-4H-cyclopenta[d][1,3]dioxan-4-yl)-5,6,7,8-tetrahydrocyclopenta[4,5]pyrrole And[2,3-d]pyrimidin-4-yl)benzamide (33.3mg, 0.067mmol) was dissolved in ammonia water/ethanol (20mL/4mL), and the reaction system was sealed and heated to 200°C in a stuffy tank and stirred for 40 Hour.
  • Step G At room temperature, N 7 -(((3aR,4R,6R,6aS)-6-(4-amino-6,7-dihydrocyclopentadieno[4,5]pyrrolo[2 ,3-d]pyrimidin-8(5H)-yl)-2,2-dimethyltetrahydro-4H-cyclopentadiene[d][1,3]dioxan-4-yl)methanol yl)quinoline-2,7-diamine (35 mg, 0.07 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A 4-Chloro-6,7-dihydro-5H-pyrrolo[2,3-D]pyrimidine (775 mg, 5.0 mmol) was dissolved in tetrahydrofuran (40 mL) at room temperature. Potassium tert-butoxide (559 mg, 5.0 mmol) was added portionwise to the reaction system at room temperature. The reaction solution was stirred at room temperature for 2 hours.
  • reaction solution was a white turbid liquid.
  • the reaction solution was concentrated, spin-dried, N,N-diisopropylethylamine (20 mL) was added, and the mixture was sonicated for 10 minutes.
  • the reaction solution was filtered, and the filter cake was 4-chloro-5,6-dihydropyrrolo[2,3-d]pyrimidine-7-potassium (wet weight: 1.1 g).
  • Step B Dissolve 4-chloro-5,6-dihydropyrrolo[2,3-d]pyrimidine-7-potassium (562 mg, 2.9 mmol) and N,N-dimethylformaldehyde at room temperature amide (12 mL). (3aR,4S,6R,6aR)-2,2-Dimethyl-6-vinyltetrahydro-4H-cyclopentadien[d][1,3]dioxan-4-yltri Fluoromethanesulfonate (910 mg, 2.9 mmol) was dissolved in N,N-dimethylformamide (6 mL). Subsequently, the above solution was slowly added to the reaction system under an ice bath. The reaction solution was stirred overnight in an oil bath at 25°C.
  • Step C At room temperature, 4-chloro-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d] [1,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (150 mg, 0.5 mmol), 7-bromoquinolin-2-amine ( 125 mg, 0.6 mmol) and tetraethylammonium chloride (85.1 mg, 0.5 mmol) were dissolved in N,N-dimethylformamide (6 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step D At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-chloro-5,6-dihydro-7H-pyrrolo[2,3- d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)quinoline-2- Amine (237 mg, 0.5 mmol), tert-butyl carbamate (240 mg, 2.1 mmol) was dissolved in N,N-dimethylformamide (5 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Tris(dibenzylideneacetone)dipalladium 47 mg, 0.05 mmol
  • 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl 25 mg, 0.05 mmol
  • cesium carbonate 333 mg, 1.0 mmol
  • Step E At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)quinoline- 2-Amine (60 mg, 0.1 mmol) was dissolved in methanol (2 mL). Subsequently, 4 mol/L hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Compound 13-2 was prepared by referring to the method of compound 1-6.
  • Step A Add dropwise to a solution of potassium 4-chloropyrrolo[2,3-d]pyrimidine-7-amide (416mg, 2.16mmol) in N,N-dimethylformamide (2mL) in an ice bath (3aR,6R,6aR)-2,2-Dimethyl-6-vinyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yltrifluoromethanesulfonyl Ester (685mg, 2.16mmol, preparation method reference compound 1-6) in N,N-dimethylformamide (2mL) solution. After the dropwise addition, the reaction solution was stirred overnight at room temperature.
  • Step B At room temperature, 4-chloro-7-((3aS,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1 ,3]dioxan-4-yl)-7H-pyrrolo[2,3-d]pyrimidine (330mg, 1.03mmol), 7-bromoquinolin-2-amine (250mg, 1.13mmol) and tetraethyl Ammonium chloride (188 mg, 1.13 mmol) was dissolved in N,N-dimethylformamide (2 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step C In a 10 mL sealed tube, add 7-((E)-2-((3aR,4R,6aS)-6-(4-chloro-7H-pyrrolo[2,3-d]pyrimidine- 7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)quinolin-2-amine (130mg , 0.28mmol), concentrated ammonia (4mL) and isopropanol (2mL). The reaction was sealed, then heated to 110°C and stirred overnight at this temperature.
  • Step D In an autoclave, add the compound 7-((E)-2-((3aR,4R,6aS)-6-(4-amino-7H-pyrrolo[2,3-d]pyrimidine-7- base)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)quinolin-2-amine (35mg, 0.079 mmol), 10 wt% palladium on carbon (10 mg) and ethanol (20 mL). The reaction system was replaced by a hydrogen atmosphere, and the pressure was 15 atmospheres and heated to 35°C. After 24 hours of reaction, the reaction was monitored, and it was found that 20% (mol fraction) of the product and 80% (mol fraction) of the raw material were present. Stirring was then continued at this pressure and temperature for two days.
  • Step E At room temperature, the compound 7-(2-((3aR,4S,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidine- 7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)ethyl)quinolin-2-amine (9mg , 0.02mmol) was dissolved in methanol (0.5mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Embodiment 13-P1 is a diagrammatic representation of Embodiment 13-P1:
  • HPLC retention time 5.327 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Embodiment 13-P2 is a diagrammatic representation of Embodiment 13-P2:
  • HPLC retention time 5.691 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Step A At room temperature, 4-chloro-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d] [1,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (630mg, 1.96mmol), tert-butyl carbamate (1.15g, 9.82 mmol) was dissolved in N,N-dimethylformamide (10 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step B At room temperature, 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-6,7-dihydro-5H-pyrrolidin[2,3-d]pyrimidin-4-amine (57mg, 0.19mmol), 3-bromo-7-iodoquinoline- 2-Amine (79 mg, 0.23 mmol) and tetraethylammonium chloride (34.6 mg, 0.21 mmol) were dissolved in N,N-dimethylformamide (2 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step C Compound 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2, 3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)-3- Bromoquinolin-2-amine (20 mg, 0.04 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A In a three-neck flask, add 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1 ,3]Dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (50 mg, 0.164 mmol) and tetrahydrofuran (1 mL). Replace the reaction system with a nitrogen system, then add 9-BBN (1.64mmol) in tetrahydrofuran (0.5M, 3.2mL) dropwise to the reaction system.
  • reaction solution is heated to 60°C and reacted for 1 hour.
  • the reaction was also cooled to room temperature, and a solution of potassium phosphate (174 mg, 0.82 mmol) in water (2 mL) was added dropwise to the reaction solution.
  • the reaction mixture was stirred at room temperature for 30 minutes, and then 3-bromo-7-iodoquinolin-2-amine (60 mg, 0.17 mmol) and [1,1'-bis(diphenylphosphino) were added to the reaction system Ferrocene]palladium dichloride (10 mg, 0.014 mmol). Replace the reaction system with nitrogen system again, then heat to 60°C and stir overnight.
  • Step B At room temperature, 7-(2-((3aR,4S,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidine -7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxy-4-yl)ethyl)-3-bromoquinoline-2 -
  • the amine 55 mg, 0.104 mmol
  • 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-chloro-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)quinoline- 2-Amine (300 mg, 0.65 mmol), di-tert-butyl dicarbonate (0.6 mL, 2.6 mmol) and 4-dimethylaminopyridine (16 mg, 0.13 mmol) were dissolved in tetrahydrofuran (10 mL). Subsequently, triethylamine (0.36 mL, 2.6 mmol) was added to the above solution. The reaction was stirred at room temperature for 18 hours.
  • Step B Slowly add potassium hydroxide (1.92g, 30mL water) aqueous solution to diethyl ether (70mL) at -10°C, then add 1-methyl-1-nitrosourea (3.5g, 33.9mmol) The reaction was stirred at -10°C for 1 hour. The reaction solution was cooled to -78°C, the aqueous phase was frozen and the organic phase was poured out. The organic phase was first washed with saturated brine (100 mL), then dried with anhydrous sodium sulfate, and filtered to obtain the diazomethane diethyl ether solution prepared now. (50mL).
  • Step C At room temperature, di-tert-butyl (7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-chloro-5,6-dihydro-7H-pyrrole And[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadiene[d][1,3]dioxan-4-yl)ring Propyl) quinoline-2-carbamate (160mg, 0.24mmol), tert-butyl carbamate (139mg, 1.18mmol) was dissolved in N,N-dimethylformamide (2mL). The reaction system was evacuated Change nitrogen several times.
  • Step D At room temperature, the compound di-tert-butyl (7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H- Pyrrolo[2,3-d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl) Cyclopropyl) quinoline-2-carbamate (62mg, 0.094mmol) was dissolved in methanol (1mL). Subsequently, 4M hydrochloric acid-methanol solution (3mL) was added to the above solution. The reaction solution was stirred at room temperature 2 hours.
  • Purification conditions are as follows, preparative column welch XBC18, 21.2 ⁇ 150mm, 5 ⁇ m (particle size); mobile phase: water (containing 10 mmol/L ammonium bicarbonate) and acetonitrile; flow rate: 20 ml/min; gradient: within 10 minutes , Acetonitrile rose from 40vol% to 60vol%; detection wavelength: 214nm.
  • HPLC retention time 5.495 minutes.
  • the conditions are as follows, column XBridge BEH C18 2.5 ⁇ m, 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, within 9 minutes Acetonitrile was raised to 95 vol%, and the 95 vol% gradient was maintained for 2 minutes.
  • HPLC retention time 3.380 minutes. Conditions are as follows, column ACQUITY UPLC BEH C18 1.7 ⁇ m, 2.1 * 150mm; Column temperature 30 °C; Mobile phase water (0.05vol% TFA) and acetonitrile (0.05vol% TFA); Flow velocity 0.3 milliliters/minute; Gradient: 5vol% acetonitrile ( 0.05vol% TFA) for 0.5 minutes, acetonitrile (0.05vol% TFA) was raised to 95vol% in 9 minutes, and the 95vol% gradient was maintained for 3 minutes.
  • Step A At room temperature, 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3] Dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (50.0 mg, 0.17 mmol), 3-bromo-N-(cyclopropyl (methyl)-7-iodoquinolin-2-amine (69.6mg, 0.17mmol) and tetraethylammonium chloride (30.4mg, 0.18mmol) were dissolved in N,N-dimethylformamide (3mL) .
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (7.5 mg, 0.03 mmol), N,N-diisopropylethylamine (107.7 mg, 0.83 mmol) were added under nitrogen atmosphere. The reaction solution was stirred in an oil bath at 70°C for 18 hours.
  • Step B At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)-3- Bromo-N-(cyclopropylmethyl)quinolin-2-amine (30 mg, 0.05 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)-3- Bromoquinolin-2-amine (160.0 mg, 0.31 mmol), di-tert-butyl dicarbonate (434.9 mg, 1.99 mmol) and 4-dimethylaminopyridine (7.6 mg, 0.06 mmol) were dissolved in tetrahydrofuran (8 mL). Subsequently, triethylamine (250.9 mg, 2.5 mmol) was added to the above solution. The reaction was stirred at room temperature for 18 hours.
  • Step B Slowly add potassium hydroxide (1.7g, 25mL water) aqueous solution into diethyl ether (60mL) at -10°C, then add 1-methyl-1-nitrosourea (3.0g, 29.1mmol) The reaction was stirred at -10°C for 1 hour. The reaction solution was cooled to -78°C, the aqueous phase was frozen and the organic phase was poured out. The organic phase was first washed with saturated brine (100 mL), then dried with anhydrous sodium sulfate, and filtered to obtain the diazomethane diethyl ether solution prepared now. (50mL).
  • Step C At room temperature, di-tert-butyl (7-((3aS,4R,6R,6aR)-6-((1R)-2-(2-(di-tert-butoxycarbonyl)amino)-3- Bromoquinolin-7-yl)cyclopropyl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-6, 7-Dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-yl)iminodicarbonate (70 mg, 0.07 mmol) was dissolved in dichloromethane (2 mL). Subsequently, trifluoroacetic acid (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • HPLC retention time 6.532 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Embodiment 18-P2 is a diagrammatic representation of Embodiment 18-P2:
  • HPLC retention time 6.676 minutes. Conditions are as follows, column XBridge BEH C18 2.5 ⁇ m 4.6mm ⁇ 150mm; column temperature 30°C; mobile phase water (10mM NH 4 HCO 3 ) and acetonitrile; flow rate 1 ml/min; gradient: 5vol% acetonitrile for 0.5 minutes, acetonitrile within 9 minutes Rise to 95vol%, hold 95vol% gradient for 2 minutes.
  • Step A In a three-necked flask, add cuprous iodide (5.92g, 31.08mmol) and anhydrous tetrahydrofuran (100mL). The replacement reaction system is a nitrogen system, and then cooled to 0°C. Methyllithium (59.7 mmol) in tetrahydrofuran (1.6 M, 37.3 mL) was added dropwise to the reaction system. After the addition was complete, stirring was continued at this temperature for 10 minutes, and then the reaction system was cooled to -78°C.
  • Step B In a three-necked flask, add (3aR,6S,6aR)-2,2,6-trimethyldihydro-4H-cyclopentadiene[d][1,3]dioxane- 4(5H)-ketone (500mg, 2.94mmol) and triethylchlorosilane (1mL, 5.88mmol) were dissolved in tetrahydrofuran (8mL), and then the system was replaced with nitrogen atmosphere. The reaction flask was cooled to -78°C in an ice bath, and then a solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1M, 4.41 mL, 4.41 mmol) was added dropwise. After the dropwise addition, the reaction system continued to stir at 0° C. for 20 minutes.
  • Step C In a one-necked bottle, add triethyl(3aR,6S,6aR)-2,2,6-trimethyl-6,6a-dihydro-4H-cyclopenta[d][1 ,3] dioxan-4-yl)oxy)silane (660mg, 2.32mmol), palladium acetate (155mg, 0.696mmol) and dimethyl sulfoxide (10mL), and then the system was replaced with an oxygen atmosphere. The reaction solution was heated to 65°C and reacted overnight at this temperature.
  • reaction solution was poured into ice water.
  • the reaction solution was extracted with ethyl acetate (30 mL ⁇ 3 times), the organic phases were combined, washed with water (30 mL ⁇ 3 times) and saturated brine (30 mL) successively, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.
  • the resulting mixture was purified by silica gel column chromatography to obtain (3aR,6aR)-2,2,6-trimethyl-4H-cyclopentadieno[d][1,3]dioxacyclo-4(6aH)- Ketones (268 mg).
  • Step D In a three-necked flask, add lithium chloride (20mg, 0.476mmol), cuprous iodide (45mg, 0.238mmol) and tetrahydrofuran (4mL), and then replace the system with nitrogen atmosphere.
  • the reaction solution was cooled to 0°C and stirred at this temperature for 10 minutes, then trimethylchlorosilane (308.5mg, 2.85mmol) and (3aR,6aR)-2,2,6- Trimethyl-4H-cyclopenta[d][1,3]dioxacyclo-4(6aH)-one (400 mg, 2.38 mmol) in tetrahydrofuran (2 mL).
  • reaction solution was stirred at 0° C. for 20 minutes.
  • a solution of vinylmagnesium bromide (3.8 mmol) in tetrahydrofuran (1M, 3.8 mL) was then added dropwise.
  • the resulting reaction solution was stirred at 0°C for 30 minutes.
  • Step E Add (3aR,6R,6aR)-2,2,6-trimethyl-6-vinyldihydro-4H-cyclopentadiene[d][1,3]dihydro to a three-neck flask A solution of oxetane-4(5H)-one (700 mg, 3.57 mmol) in tetrahydrofuran (10 mL), and then the system was replaced with a nitrogen atmosphere. The reaction flask was placed in an ice bath and cooled to 0° C., and then a solution of lithium aluminum hydride (7.14 mmol) in tetrahydrofuran (1M, 7.14 mL) was added dropwise. After the dropwise addition, the reaction system continued to stir at 0° C. for 1 hour.
  • Step F 4-Chloro-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (275 mg, 1.77 mmol) and (3aS,4S,6R,6aR)-2, 2,6-Trimethyl-6-vinyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-ol (440mg, 2.22mmol) was dissolved in toluene (5mL) , the reaction system was replaced by a nitrogen atmosphere. Then a solution of 2-(tributylphosphoranylidene)acetonitrile (853 mg, 3.54 mmol) in toluene (2 mL) was slowly added to the reaction system. The reaction solution was heated to 110°C and stirred at this temperature for 5 hours.
  • Step G At room temperature, add 4-chloro-7-((3aS,4R,6R,6aR)-2,2,6-trimethyl-6-vinyltetrahydro-4H-ring to the autoclave Pentadieno[d][1,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (130 mg, 0.39 mmol) in ethanol ( 3mL) solution. Subsequently, aqueous ammonia (30 mL) was slowly added to the reaction system at room temperature. The reaction solution was stirred in an oil bath at 135° C. for 2 days.
  • Step H 7-((3aS,4R,6R,6aR)-2,2,6-trimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1 ,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (80mg, 0.25mmol), 3-bromo-7-iodo Quinolin-2-amine (88 mg, 0.25 mmol) and tetraethylammonium chloride (45.6 mg, 0.28 mmol) were dissolved in N,N-dimethylformamide (1 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step I At room temperature, 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2,4-trimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)vinyl)- 3-Bromoquinolin-2-amine (28 mg, 0.05 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Purification conditions are as follows, preparative column welch XBC18, 21.2 ⁇ 150mm, 5 ⁇ m (particle size); mobile phase: water (containing 10 mmol/L ammonium bicarbonate) and acetonitrile; flow rate: 20 ml/min; gradient: within 10 minutes , Acetonitrile rose from 40vol% to 60vol%; detection wavelength: 214nm.
  • Step A In a three-necked flask, add cuprous bromide dimethyl sulfide complex (411 mg, 2 mmol) and tetrahydrofuran (80 mL). The replacement reaction system is a nitrogen system, and then cooled to -78°C.
  • Step B Add (6S)-6-allyl-2,2-dimethyldihydro-4H-cyclopentadieno[d][1,3]dioxane-4 into a three-neck flask (5H)-Kone (1 g, 5.1 mmol) was dissolved in tetrahydrofuran (30 mL), and then the system was replaced with a nitrogen atmosphere. The reaction flask was cooled to 0° C. in an ice bath, and then a solution of lithium aluminum hydride (10 mmol) in tetrahydrofuran (1M, 10 mL) was added dropwise. After the dropwise addition, the reaction system continued to stir at 0° C. for 1 hour.
  • Step C At room temperature, mix 4-chloro-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (104mg, 0.67mmol) and (4S,6S)-6-allyl- 2,2-Dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-ol (200 g, 1.01 mmol) was dissolved in toluene (2 mL). 2-(Tributylphosphoranylidene)acetonitrile (482 mg, 2.0 mmol) was dissolved in toluene (0.5 mL). Subsequently, 2-(tributylphosphoranylidene)acetonitrile solution was slowly added to the reaction system at room temperature. The reaction solution was stirred for 12 hours in an oil bath at 110°C.
  • Step D At room temperature, add 7-((3aS, 4R, 6S, 6aR)-6-allyl-2,2-dimethyltetrahydro-4H-cyclopentadiene to the autoclave [ d][1,3]dioxan-4-yl)-4-chloro-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (244mg, 0.73mmol) in ethanol (3mL ) solution. Subsequently, aqueous ammonia (25 mL) was slowly added to the reaction system at room temperature. The reaction solution was stirred in an oil bath at 135° C. for 2 days.
  • Step E 7-((3aS,4R,6S,6aR)-6-allyl-2,2-dimethyltetrahydro-4H-cyclopentadiene[d][1, 3] Dioxacyclo-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (70mg, 0.22mmol), 3-bromo-7-iodoquinone Phenyl-2-amine (77 mg, 0.22 mmol) and tetraethylammonium chloride (40 mg, 0.24 mmol) were dissolved in N,N-dimethylformamide (1 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step F At room temperature, 7-((E)-3-((3aR,4S,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3 -d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)prop-1-ene- 1-yl)-3-bromoquinolin-2-amine (62 mg, 0.12 mmol) was dissolved in methanol (0.5 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 4-chloro-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d] [1,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (210 mg, 0.65 mmol), 1,1'-bis(diphenyl Phosphino)ferrocenepalladium(II) chloride (95.1 mg, 0.13 mmol) and potassium carbonate (179.6 mg, 1.3 mmol) were dissolved in dioxane/water (5 mL/0.5 mL). The reaction system was evacuated and replaced with nitrogen several times. Add 2,4,6-trimethyl-1,3,5,2,4,6-trioxaborohexane (246.9mg, 1.96mmol) under nitrogen atmosphere, seal the reaction system and heat to 110°C and stirred for 18 hours.
  • Step B At room temperature, 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-4-methyl-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine (40mg, 0.13mmol) and 3-bromo-7-iodoquinone Phenyl-2-amine (42.1 mg, 0.12 mmol) was dissolved in N,N-dimethylformamide (2 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step C At room temperature, 3-bromo-7-((E)-2-((3aR,4R,6R,6aS)-2,2-dimethyl-6-(4-methyl-5, 6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)tetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)ethylene yl)quinolin-2-amine (36.0 mg, 0.07 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A Add 7-((3aS,4R,6R,6aR)-2,2,6-trimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][ 1,3]dioxan-4-yl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidin-4-amine (45 mg, 0.14 mmol) in THF (0.5 mL) , and then the system was replaced with a nitrogen atmosphere. At room temperature, a tetrahydrofuran solution (0.5M, 2.8 mL) of 9-BBN (1.4 mmol) was slowly added dropwise to the reaction solution. After the addition was complete, the reaction solution was heated to 60° C.
  • Step B At room temperature, 7-(2-((3aR,4S,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidine -7-yl)-2,2,4-trimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)ethyl)-3-bromoquinoline Lin-2-amine (45 mg, 0.083 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • the preparation method refers to Example 21, and finally the target product (1S,2R,3R,5R)-3-((E)-2-(2-amino-3-chloroquinolin-7-yl)vinyl)-5- (4-Methyl-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopentane-1,2-diol.
  • Step A Mix 4-chloro-9H-pyrimido[4,5-b]indole (450mg, 2.2mmol) and (3aS,4S,6S,6aR)-2,2-dimethyl- A mixture of 6-allyltetrahydro-4H-cyclopenta[d][1,3]dioxan-4-ol (806 mg, 4.4 mmol) in toluene (5 mL) was added to a sealed 20 mL tube .
  • reaction mixture was replaced with a nitrogen system, and then, a solvent of 2-(tributylphosphoranylidene)acetonitrile (2.12 g, 8.8 mmol) in toluene (3 mL) was slowly added dropwise to the reaction system at room temperature.
  • the reaction solution was stirred for 12 hours in an oil bath at 100°C.
  • Step B Add 4-chloro-9-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H- Cyclopenta[d][1,3]dioxan-4-yl)-9H-pyrimido[4,5-b]indole (360 mg, 0.98 mmol) in isopropanol (3 mL) . Subsequently, aqueous ammonia (6 mL) was slowly added to the reaction system at room temperature. The tube was sealed, and the reaction solution was stirred overnight in an oil bath at 120°C.
  • Step C Add 9-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1, 3] Dioxan-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (50 mg, 0.14 mmol) in tetrahydrofuran (0.5 mL), and then the system was replaced with nitrogen atmosphere. At room temperature, a tetrahydrofuran solution (0.5M, 2.8 mL) of 9-BBN (1.4 mmol) was slowly added dropwise to the reaction solution. After the addition was complete, the reaction solution was heated to 60° C. and stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, then an aqueous solution (0.5 mL) of potassium phosphate (148 mg, 0.7 mmol) was added to the reaction mixture. After the reaction solution was stirred at room temperature for 30 minutes, a solution of 3-bromo-7-iodoquinolin-2-amine (50 mg, 0.14 mmol) and PdCl 2 (dppf) (10 mg, 0.014 mmol) in tetrahydrofuran (1.2 mL) was added. The resulting reaction mixture was heated to 60°C and stirred overnight.
  • Step D At room temperature, 9-((3aS,4R,6S,6aR)-6-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-2,2-di Methyltetrahydro-4H-cyclopentadieno[d][1,3]dioxol-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (70mg, 0.12 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A 9-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (50mg, 0.14mmol), 3-bromo-7-iodoquinolin-2-amine (60.0mg , 0.17mmol) and tetraethylammonium chloride (26.0mg, 0.16mmol) were dissolved in N,N-dimethylformamide (3mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step B At room temperature, 9-((3aS,4R,6R,6aR)-6-((E)-2-(2-amino-3-bromoquinolin-7-yl)ethenyl)-2 ,2-Dimethyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (35 mg, 0.06 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A 9-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentadiene[d][1,3 ]dioxan-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (100mg, 0.28mmol), 7-iodoquinolin-2-amine (91.8mg, 0.34mmol) and tetraethylammonium chloride (51.8 mg, 0.31 mmol) were dissolved in N,N-dimethylformamide (3 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step B Dissolve palladium on carbon (236.0 mg, 2.2 mmol) in ethanol (5 mL) at room temperature.
  • the reaction system was evacuated and hydrogen was exchanged several times.
  • the reaction was stirred at room temperature for 18 hours.
  • Step C At room temperature, 9-((3aS,4R,6S,6aR)-6-(2-(2-aminoquinolin-7-yl)ethyl)-2,2-dimethyltetrahydro -4H-cyclopentadieno[d][1,3]dioxan-4-yl)-9H-pyrimido[4,5-b]indol-4-amine (70mg, 0.14mmol) was dissolved in in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, in a single-necked bottle, add acetonitrile (50mL), 4-amino-6-chloropyrimidin-5-ol (437mg, 3mmol) and cesium carbonate (1955mg, 6mmol), 1,2-dibromo Ethane (1127mg, 6mmol), then heated to 65°C and stirred for 16 hours.
  • Step B In a locked tube, mix 4-chloro-7,8-dihydro-6H-pyrimidine[5,4-b][1,4]oxazine (205 mg, 1.2 mmol) and (3aS, 4S, 6R ,6aR)-2,2,6-Trimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]dioxin-4-ol (442mg, 2.4mmol) in toluene (3 mL).
  • Cyanomethylenetri-n-butylphosphine (869mg, 3.6mmol) was dissolved in toluene (1mL), and the above liquid was added dropwise under nitrogen atmosphere, and the reaction solution was stirred at 110°C for 18 hours.
  • Step C 4-chloro-8-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1 ,3]dioxol-4-yl)-7,8-dihydro-6H-pyrimidine[5,4-b][1,4]oxazine (295 mg, 0.872 mmol), tert-butyl carbamate (202.5 mg, 1.74mmol) was dissolved in anhydrous toluene (2mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Brettphos Pd G2 (73.6 mg, 0.087 mmol)
  • cesium carbonate (566.8 mg, 1.74 mmol) were added under nitrogen atmosphere.
  • the reaction solution was stirred at 110°C for 18 hours.
  • reaction solution was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography to obtain 8-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d ][1,3]dioxol-4-yl)-7,8-dihydro-6H-pyrimidin[5,4-b][1,4]oxazin-4-amine (105 mg).
  • Step D In a three-necked flask, under nitrogen protection, add anhydrous tetrahydrofuran (5mL), 8-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H- Cyclopentane[d][1,3]dioxol-4-yl)-7,8-dihydro-6H-pyrimidin[5,4-b][1,4]oxazin-4-amine (115mg , 0.35mmol), then 9-BBN (299mg, 2.45mmol) was added dropwise. The reaction solution was heated to 55°C and stirred for 1 hour.
  • 5mL anhydrous tetrahydrofuran
  • the obtained crude product was purified by silica gel column chromatography to obtain 8-((3aS, 4R, 6S, 6aR)-6-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-2,2- Dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)-7,8-dihydro-6H-pyrimidine[5,4-b][1,4] Oxazin-4-amine (105 mg).
  • Step E At room temperature, in a single-necked flask, add methanol (2mL), 8-((3aS, 4R, 6S, 6aR)-6-(2-(2-amino-3-bromoquinolin-7-yl )ethyl)-2,2-dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)-7,8-dihydro-6H-pyrimidine[5, 4-b] [1,4]oxazin-4-amine (81mg, 0.15mmol), was added dropwise with 4N methanolic hydrochloric acid solution (2mL), then stirred for 2 hours.
  • Step A In a one-necked flask, add anhydrous DMF (2mL), 8-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[ d][1,3]dioxol-4-yl)-7,8-dihydro-6H-pyrimidin[5,4-b][1,4]oxazin-4-amine (63mg, 0.2mmol) , 3-bromo-7-iodoquinolin-2-amine (84 mg, 0.24 mmol), palladium acetate (10 mg, 0.04 mmol), TEAC (66 mg, 0.4 mmol), DIPEA (51 mg, 0.394 mmol).
  • Step B At room temperature, in a one-necked flask, add methanol (2mL), 8-((3aS, 4R, 6R, 6aR)-6-((E)-2-(2-amino-3-bromoquinoline -7-yl)vinyl)-2,2-dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)-7,8-dihydro-6H- Pyrimidin[5,4-b][1,4]oxazin-4-amine (25mg, 0.05mmol) was added dropwise with 4N methanolic hydrochloric acid solution (2mL), then stirred for 2 hours.
  • the preparation method refers to Example 27, and finally the target product (1S,2R,3S,5R)-3-(2-(2-amino-3-bromoquinolin-7-yl)ethyl)-5-(4- Amino-6,7-dihydro-8H-pyrimidin[5,4-b][1,4]oxazin-8-yl)-3-methylcyclopentane-1,2-diol.
  • Step A 8-((3aS,4R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]dioxol -4-yl)-5,6,7,8-tetrahydrocyclopentane[4,5]pyrrole[2,3-d]pyrimidin-4-amine (53mg, 0.156mmol), 7-bromo-3- Chloroquinolin-2-amine (40mg, 0.156mmol, prepared with reference to the synthesis method of intermediate Int-1) and tetraethylammonium chloride (52.2mg, 0.315mmol) were dissolved in N,N-dimethylformamide (1mL )middle.
  • the reaction system was evacuated and replaced with nitrogen several times. Palladium acetate (7 mg, 0.03 mmol), N,N-diisopropylethylamine (40 mg, 0.312 mmol) were added under nitrogen atmosphere. The reaction solution was stirred for 12 hours in an oil bath at 100°C.
  • Step B At room temperature, 8-((3aS, 4R, 6R, 6aR)-6-((E)-2-(2-amino-3-chloroquinolin-7-yl)ethenyl)-2, 2-Dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)-5,6,7,8-tetrahydrocyclopentane[4,5]pyrrole[ 2,3-d]pyrimidin-4-amine (63 mg, 0.122 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A 7-Bromo-3-fluoroquinoline (900 mg, 4 mmol) was dissolved in dichloromethane (25 mL) at room temperature. m-chloroperbenzoic acid (2.75 g, 16 mmol) was slowly added at 0° C., and the reaction solution was stirred at room temperature for 12 hours.
  • Step B 7-Bromo-3-fluoroquinoline 1-oxide (850 mg, 3.54 mmol) was dissolved in chloroform (15 mL) at room temperature. Then, phosphorus oxychloride (15 mL) was slowly added at 0° C., and the reaction system was evacuated and replaced with nitrogen several times. The reaction solution was stirred for 3 hours in an oil bath at 70°C.
  • Step C 7-Bromo-2-chloro-3-fluoroquinoline (745 mg, 2.86 mmol) was dissolved in ethanol (10 mL) and ammonia (20 mL) at room temperature. Then the reaction solution was placed in a stewing tank and stirred at 100° C. for 20 hours.
  • Step D 8-((3aS,4R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]dioxol -4-yl)-5,6,7,8-tetrahydrocyclopentane[4,5]pyrrole[2,3-d]pyrimidin-4-amine (125mg, 0.36mmol), 7-bromo-2- Chloro-3-fluoroquinoline (86 mg, 0.36 mmol) and tetraethylammonium chloride (65 mg, 0.39 mmol) were dissolved in N,N-dimethylformamide (2 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step E At room temperature, 8-((3aS,4R,6R,6aR)-6-((E)-2-(2-amino-3-fluoroquinolin-7-yl)ethenyl)-2 ,2-Dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)-5,6,7,8-tetrahydrocyclopentane[4,5]pyrrole [2,3-d]Pyrimidin-4-amine (87 mg, 0.174 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (3 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • reaction solution was concentrated under reduced pressure to obtain a crude product.
  • the crude product was dissolved in 2 mL methanol, and then 7M ammonia methanol solution was added to adjust the pH value to approximately equal to 8-9.
  • the preparation method refers to Example 31, and finally the target product (1S,2R,3R,5R)-3-((E)-2-(2-amino-3-fluoroquinolin-7-yl)ethenyl)-5 -(4-Amino-5,6-dihydro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)cyclopentane-1,2-diol.
  • Step A At room temperature, 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]di Oxyalcohol-4-yl)-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidin-4-amine (60mg, 0.2mmol), 7-bromo-3-chloroquinolin-2-amine ( 51 mg, 0.2 mmol) and tetraethylammonium chloride (36.3 mg, 0.22 mmol) were dissolved in N,N-dimethylformamide (1.5 mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step B 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrole[2,3- d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)vinyl)-3-chloroquinoline-2 -
  • the amine 40 mg, 0.084 mmol
  • 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution.
  • the reaction solution was stirred at room temperature for 2 hours.
  • Step A In a three-necked flask, a solution of methyl 2-(4,6-dichloropyrimidin-5-yl)acetate (2 g, 9.1 mmol) in tetrahydrofuran (100 mL) was added.
  • the replacement reaction system is a nitrogen system, and then cooled to -78°C.
  • Add a tetrahydrofuran solution (1M, 18.2mL) of lithium bistrimethylsilylamide (18.2mmol) dropwise to the reaction system, continue stirring at this temperature for 1 hour after the addition is complete, and then drop Iodomethane (2.58 g, 18.2 mmol) was added. After the dropwise addition, the reaction system was raised to 0° C. and stirred for 2 hours.
  • Step B In a three-necked flask, add a solution of methyl 2-(4,6-dichloropyrimidin-5-yl)propionate (1 g, 4.27 mmol) in tetrahydrofuran (30 mL).
  • the replacement reaction system is a nitrogen system, and then cooled to minus 78°C.
  • a tetrahydrofuran solution (1.5 M, 8.5 mL) of diisobutylaluminum hydride (12.8 mmol) was added dropwise to the reaction system. After the addition was complete, the reaction system was slowly raised to room temperature and continued to stir overnight.
  • Step C In a three-necked flask, 2-(4,6-dichloropyrimidin-5-yl)propan-1-ol (540mg, 2.62mmol) and 4-dimethylaminopyridine (32mg, 0.262mmol) were dissolved in in dichloromethane (420 mL). Methanesulfonyl chloride (750mg, 6.55mmol) and triethylamine (1.19g, 11.8mmol) were added at 0°C, and the reaction solution was stirred at 0°C to room temperature for 3 hours.
  • Step D At room temperature, mix 2-(4,6-dichloropyrimidin-5-yl)propyl methanesulfonate (700mg, 2.46mmol) and (4-methoxyphenyl)formamide (1.04g , 7.4mmol) was dissolved in dichloromethane (30mL), then triethylamine (749mg, 7.4mmol) was added to replace the reaction system with nitrogen system, and the reaction solution was stirred overnight at 55°C.
  • Step E At room temperature, add 4-chloro-7-(4-methoxybenzyl)-5-methyl-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (710mg, 2.46 mmol) was dissolved in trifluoroacetic acid (3 mL) and trifluoromethanesulfonic acid (0.3 mL), and the reaction solution was stirred at 70° C. for 3 hours.
  • Step F In a sealed tube, mix 4-chloro-5-methyl-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (460mg, 2.72mmol) and (3aS,4S,6R, 6aR)-2,2-Dimethyl-6-vinyltetrahydro-4H-cyclopentadieno[d][1,3]dioxan-4-ol (763.5 mg, 4.08 mmol, ref. J.Org.Chem.2004, 69, 3993-3996 method, the entire content of which is incorporated in this application by reference) was dissolved in toluene (2 mL).
  • Cyanomethylenetri-n-butylphosphine (1.31 g, 5.44 mmol) was dissolved in toluene (2 mL), and the above liquid was added dropwise under a nitrogen atmosphere, and the reaction liquid was stirred at 110° C. for 18 hours.
  • Step G 4-chloro-7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1 ,3]dioxol-4-yl)-5-methyl-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (300 mg, 0.895 mmol) was dissolved in ethanol (20 mL) and ammonia ( 100 mL) of the mixed solution, and the reaction solution was stirred in a 160°C autoclave for 60 hours.
  • Step H Add compound 7-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane [d][1,3 in a three-necked flask ]dioxol-4-yl)-5-methyl-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidin-4-amine (38 mg, 0.11 mmol) in tetrahydrofuran (4 mL), Then the system was replaced with a nitrogen atmosphere. At room temperature, 9-BBN (1.1 mmol) in tetrahydrofuran (0.5 M, 2.2 mL) was slowly added dropwise to the reaction solution.
  • reaction solution was heated to 55° C. and stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, and then an aqueous solution (1 mL) of potassium phosphate (233 mg, 1.1 mmol) was added to the reaction mixture.
  • an aqueous solution (1 mL) of potassium phosphate 233 mg, 1.1 mmol was added to the reaction mixture.
  • a solution of 3-bromo-7-iodoquinolin-2-amine 38.5 mg, 0.11 mmol
  • PdCl2 (dppf) 8 mg, 0.01 mmol
  • Step 1 At room temperature, 7-(2-((3aR, 4S, 6R, 6aS)-6-(4-amino-5-methyl-5H-pyrrole[2,3-d]pyrimidine-7( 6H)-yl)-2,2-dimethyltetrahydro-3aH-cyclopentane[d][1,3]dioxy-4-yl)ethyl)-3-bromoquinolin-2-amine (30 mg, 0.055 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A Add compound 4-chloro-7-((3aS, 4R, 6aR)-2,2-dimethyl-6-vinyl-3a, 6a-dihydro-4H-cyclopentane in a three-necked flask [d][1,3]dioxol-4-yl)-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (75mg, 0.23mmol) in tetrahydrofuran (1mL) solution, then the system Replaced with a nitrogen atmosphere. At room temperature, a solution of 9-BBN (2.35 mmol) in tetrahydrofuran (0.5 M, 4.7 mL) was slowly added dropwise to the reaction solution.
  • reaction solution was heated to 55° C. and stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, and then an aqueous solution (2 mL) of potassium phosphate (498 mg, 2.35 mmol) was added to the reaction mixture.
  • an aqueous solution (2 mL) of potassium phosphate (498 mg, 2.35 mmol) was added to the reaction mixture.
  • a solution of 3-bromo-7-iodoquinolin-2-amine 80.5 mg, 0.23 mmol
  • PdCl2 (dppf) (14.6 mg, 0.02 mmol) in THF (2 mL) was added.
  • the resulting reaction mixture was heated to 55°C and stirred overnight.
  • Step B At room temperature, 3-bromo-7-(2-((3aS, 6aR)-4-(4-chloro-5,6-dihydro-7H-pyrrole[2,3-d]pyrimidine- 7-yl)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopentane[d][1,3]dioxy-6-yl)ethyl)quinolin-2-amine (120mg, 0.22mmol) was dissolved in ethanol (5mL) and ammonia (30mL). The reaction solution was stirred for 2 days in a high-pressure reactor at 140°C.
  • Step C 7-((3aS,6aR)-2,2-Dimethyl-6-vinyl-3a,6a-dihydro-4H-cyclopentane[d][1,3] Dioxyol-4-yl)-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidin-4-amine (30 mg, 0.057 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • the preparation method refers to Example 34, and finally the target product (1S,2R,3S)-3-(2-(2-amino-3-chloroquinolin-7-yl)ethyl)-5-(4-methyl- 5,6-Dihydro-7H-pyrrole[2,3-d]pyrimidin-7-yl)cyclopentan-1,2-ol.
  • the preparation method refers to Example 35, and finally the target product (1S,2R)-3-((E)-2-(2-amino-3-bromoquinolin-7-yl)vinyl)-5-(4- Amino-5,6-dihydro-7H-pyrrole[2,3-d]pyrimidin-7-yl)cyclopent-3-ene-1,2-diol.
  • Step A Add compound 4-chloro-7-((3aS, 4R, 6aR)-2,2-dimethyl-6-vinyl-3a, 6a-dihydro-4H-cyclopentane in a three-necked flask [d][1,3]dioxol-4-yl)-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (110mg, 0.34mmol) in tetrahydrofuran (1mL) solution, then the system Replaced with a nitrogen atmosphere. At room temperature, a solution of 9-BBN (3.44 mmol) in tetrahydrofuran (0.5 M, 6.89 mL) was slowly added dropwise to the reaction solution.
  • reaction solution was heated to 55° C. and stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, then an aqueous solution (2 mL) of potassium phosphate (729 mg, 3.44 mmol) was added to the reaction mixture.
  • an aqueous solution (2 mL) of potassium phosphate (729 mg, 3.44 mmol) was added to the reaction mixture.
  • a solution of 7-bromo-3-chloroquinolin-2-amine (88 mg, 0.34 mmol) and PdCl 2 (dppf) (21.9 mg, 0.03 mmol) in tetrahydrofuran (2 mL) was added.
  • the resulting reaction mixture was heated to 55°C and stirred overnight.
  • Step B At room temperature, 3-chloro-7-(2-((3aS, 6aR)-4-(4-chloro-5,6-dihydro-7H-pyrrole[2,3-d]pyrimidine- 7-yl)-2,2-dimethyl-3a,6a-dihydro-4H-cyclopentane[d][1,3]dioxy-6-yl)ethyl)quinolin-2-amine (60mg, 0.12mmol) was dissolved in ethanol (5mL) and ammonia water (30mL). The reaction solution was stirred for 2 days in a high-pressure reactor at 140°C.
  • Step C At room temperature, 7-(2-((3aS,6aR)-4-(4-amino-5,6-dihydro-7H-pyrrole[2,3-d]pyrimidin-7-yl) -2,2-Dimethyl-3a,6a-dihydro-4H-cyclopentane[d][1,3]dioxy-6-yl)ethyl)-3-chloroquinolin-2-amine ( 20 mg, 0.042 mmol) was dissolved in methanol (1 mL). Subsequently, 4M hydrochloric acid-methanol solution (1 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A At room temperature, 7-((3aS,4R,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]di Oxyalcohol-4-yl)-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidin-4-amine (100mg, 0.33mmol), 7-bromoquinazolin-2-amine (81.2mg , 0.33mmol) and tetraethylammonium chloride (60.15mg, 0.363mmol) were dissolved in N,N-dimethylformamide (1.5mL). The reaction system was evacuated and replaced with nitrogen several times.
  • Step B 7-((E)-2-((3aR,4R,6R,6aS)-6-(4-amino-5,6-dihydro-7H-pyrrole[2,3- d]pyrimidin-7-yl)-2,2-dimethyltetrahydro-4H-cyclopentane[d][1,3]dioxy-4-yl)vinyl)quinazolin-2-amine (130 mg, 0.29 mmol) was dissolved in methanol (2 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A In a locked tube, combine 4-chloro-9H-pyrimido[4,5-b]indole (660mg, 3.24mmol) and (3aS,4S,6R,6aR)-2,2-dimethyl -6-vinyltetrahydro-4H-cyclopentadieno[d][1,3]dioxane-4-ol (890mg, 4.88mmol, reference J.Org.Chem.2004,69,3993 -3996 method, the entire contents of which are incorporated into this application by reference) dissolved in dry toluene (5 mL).
  • Cyanomethylene tri-n-butylphosphine (1.56 g, 6.48 mmol) was dissolved in toluene (3 mL), and the above liquid was added dropwise under nitrogen atmosphere, and the reaction liquid was stirred at 110° C. for 18 hours.
  • Step B In a sealed tube, add 4-chloro-9-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][ 1,3]dioxol-4-yl)-9H-pyrimidin[4,5-b]indole (379mg, 1.02mmol) was dissolved in ethanol (4mL) and ammonia water (10mL) mixture, and the reaction solution was heated at 120 °C and stirred for 18 hours in a sealed tube.
  • Step C Add compound 7-((3aS, 4R, 6R, 6aR)-9-((3aS, 4R, 6R, 6aR)-2,2-dimethyl-6-vinyl tetrahydro in a three-necked flask -4H-cyclopentane[d][1,3]dioxol-4-yl)-9H-pyrimidin[4,5-b]indol-4-amine (60 mg, 0.17 mmol) in THF (1 mL) solution, and then the system was replaced with a nitrogen atmosphere. At room temperature, a tetrahydrofuran solution (0.5M, 2.39mL) of 9-BBN (1.19mmol) was slowly added dropwise to the reaction solution.
  • reaction solution was heated to 55°C. Stirred at this temperature for 1 hour.
  • the reaction system was removed from the oil bath and cooled to room temperature, then an aqueous solution (2mL) of potassium phosphate (252mg, 1.19mmol) was added to the reaction mixture.
  • an aqueous solution (2mL) of potassium phosphate (252mg, 1.19mmol) was added to the reaction mixture.
  • 7-bromo-3-fluoroquinolin-2-amine 41mg, 0.17mmol, prepared by referring to the synthetic method of 7-bromo-3-chloroquinolin-2-amine in Example 31
  • PdCl 2 (dppf) (14.6mg , 0.02 mmol
  • Step D At room temperature, 9-((3aS, 4R, 6S, 6aR)-6-(2-(2-amino-3-fluoroquinolin-7-yl)ethyl)-2,2-di Methyltetrahydro-4H-cyclopentane[d][1,3]dioxol-4-yl)-9H-pyrimidin[4,5-b]indol-4-amine (60mg, 0.12mmol) in methanol (2 mL). Subsequently, 4M hydrochloric acid-methanol solution (2 mL) was added to the above solution. The reaction solution was stirred at room temperature for 2 hours.
  • Step A Add (3aR,6R,6aR)-2,2-dimethyl-6-vinyltetrahydro-4H-cyclopentane[d][1,3]dioxin-4- Ketone (550mg, 1.72mmol) in N,N-dimethylformamide (10mL) solution, and then the system was replaced with a nitrogen atmosphere.
  • the reaction flask was cooled to 0°C in an ice bath, and then potassium tert-butoxide powder (579 mg, 5.16 mmol) was added in batches. After the addition was complete, the reaction system continued to stir at 0°C for 1 hour. Then methyl iodide (732 mg, 5.16 mmol) was added dropwise under nitrogen atmosphere, and the reaction system was stirred at room temperature for 2 hours.
  • Step B Add 4-chloro-7-(2,4-dimethoxybenzyl)-5,5-dimethyl-5,7-dihydro-6H-pyrrole[2,3- -d]
  • pyrimidin-6-one 460 mg, 1.33 mmol
  • tetrahydrofuran 10 mL
  • lithium aluminum hydride 2.66 mmol
  • tetrahydrofuran 1M, 2.66 mL
  • Step C Add 4-chloro-7-(2,4-dimethoxybenzyl)-5,5-dimethyl-6,7-dihydro-5H-pyrrole[2,3- -d] A solution of pyrimidin-6-ol (370 mg, 1.06 mmol) in dichloromethane (20 mL), and then the system was replaced with a nitrogen atmosphere. The reaction flask was cooled to 0°C in an ice bath, then trifluoroacetic acid (2.3 mL) and triethylsilane (2.3 mL) were added. After the addition was complete, the reaction was stirred at room temperature for 1 hour.
  • Step D In a one-necked bottle, 4-chloro-7-(2,4-dimethoxybenzyl)-5,5-dimethyl-6,7-dihydro-5H-pyrrole[2,3 -d] Pyrimidine (330mg, 1.01mmol) was dissolved in trifluoroacetic acid (15mL), and the reaction solution was stirred overnight at 70°C.
  • Step E In a sealed tube, mix 4-chloro-5,5-dimethyl-6,7-dihydro-5H-pyrrole[2,3-d]pyrimidine (80mg, 0.44mmol) and (3aS,4S ,6R,6aR)-2,2-Dimethyl-6-vinyltetrahydro-4H-cyclopentadien[d][1,3]dioxan-4-ol (123.4mg, 0.66mmol , prepared by the method of reference J.Org.Chem.2004,69,3993-3996, the entire content of which is incorporated in this application by reference) was dissolved in dry toluene (1 mL).
  • Cyanomethylene tri-n-butylphosphine (318mg, 1.32mmol) was dissolved in toluene (0.5mL), and the above liquid was added dropwise under nitrogen atmosphere, and the reaction solution was stirred at 110°C for 18 hours.

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Abstract

一种PRMT5激酶抑制剂，含有所述化合物的药物组合物以及利用所述化合物治疗细胞增殖性疾病，例如癌症的方法。

Description

PRMT5抑制剂
本申请要求于2021年7月20日提交中国专利局、申请号为202110816281.8发明名称为“PRMT5抑制剂”的中国专利申请、2022年7月7日提交中国专利局、申请号为202210795578.5发明名称为“PRMT5抑制剂”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。
技术领域
本发明属于药物化学领域，具体涉及一种喹啉取代的PRMT5抑制剂，含有所述化合物的药物组合物以及利用本发明化合物治疗细胞增殖性疾病，例如癌症的方法。
背景技术
蛋白质精氨酸甲基转移酶(protein arginine methyltransferase，PRMTs)在蛋白质的甲基化中起着重要的作用，如参与可变剪切、转录后调节、RNA的加工、细胞增殖、细胞分化、细胞凋亡和肿瘤形成等。目前，已经鉴定出11种该家族的成员(PRMT1～11)，根据其催化精氨酸甲基化方式的不同，可以将PRMTs分为3类：I型包括PRMT1、PRMT2、PRMT3、PRMT4、PRMT6和PRMT8，催化的形式为单甲基(MMA)和不对称双甲基(aDMA)；II型为对称双甲基(sDMA)，包括PRMT5及PRMT9；III型为PRMT7。
PRMT5首次在与Janus酪氨酸激酶2(Jak2)相互作用的蛋白复合体中分离出来，所以又称为Jak结合蛋白1(JBP1)。PRMT5作为一种表观遗传酶，能对称性地甲基化组蛋白或者非组蛋白底物的精氨酸残基，影响多个靶基因及多条信号通路途径，因而发挥着多种生物学功能。研究表明，PRMT5还是一个癌基因，在多种肿瘤中高表达，且其表达水平与肿瘤的发生发展及预后密切相关。
CYP450酶参与的代谢途径是化合物在体内清除过程中非常重要的代谢途径，这一家族酶活性的抑制可带来药物体内清除和药代动力学的变化，由CYP450酶介导的药物相互作用(drug-drug interaction，DDI)是药效和药物安全性考量的重要因素(J Pharmacol Exp Ther.2006 Jan[J].2006,316(1):336-48)。CYP3A4是CYP450家族主要代谢酶，抑制CYP3A4带来的DDI会导致严重的安全性问题(The AAPS Journal volume 24,Article number:12(2022))。CYP3A4是肝脏及肠壁中含量最丰富的P450同工酶，可参与代谢约50％的临床药物，临床联合用药时，抑制或诱导CYP3A4的药物(perpetrator drug)可以影响合并用其他药物的药代动力学，从而影响血浆中药物的暴露量导致药动学DDI。抑制性(包括可逆、机制性失活)药物相互作用多提高药效，但对于治疗窗较窄的药物易引起临床不良反应，严重时可危及生命。而诱导引起的药动学DDI较少引起用药安全问题，但可降低药物的疗效。因此，预测新药可能引起的DDI是新药研发中评价候选药物特性的重要内容(张庆颢等:基于体外CYP3A4抑制和诱导数据定量预测体内药物-药物相互作用.药学学报Acta Pharmaceutica Sinica[J].2010,45(8):952-959)。
药物对P450酶的抑制通常分为可逆性抑制(包括竞争性抑制、非竞争性抑制和反竞争性抑制)和不可逆性抑制。可逆性抑制剂以非共价键与酶(或酶-底物复合物)形成复合物， 阻碍酶与底物之间酶促反应的正常进行，在除去抑制剂后，酶活性不受影响，能与底物继续进行正常酶促反应。在不可逆性抑制中，抑制剂对酶的抑制效应在除去抑制剂后不会即刻消失，而是呈现出时间依赖的特性。这种现象一般称之为时间依赖性抑制(time-dependent inhibition，TDI)。
药物对P450酶产生TDI有多种机制，其中，机理性抑制(mechanism based inhibition，MBI)是TDI最重要的机制，即抑制剂可经CYP介导转化成亲电性的反应性代谢物(reactive metabolite)，这些反应性代谢物可与酶发生作用(主要以共价键结合形式)导致酶结构的变化而失活。MBI的产生需要先经过P450酶代谢的过程，因此抑制剂对P450酶的抑制作用需要一定时间，而体内合成新的P450酶也需要一定时间(一般需要4～7d)，这样即使把抑制剂除去，其抑制作用还会存在一段时间。与可逆性抑制相比，TDI会带来更严重的用药安全问题，因为许多需联合用药的药物也需长期用药，这势必会造成被抑制的CYP亚型被长期抑制，即使停用产生TDI的药物，其抑制作用仍会持续一段时间；同时，由于产生MBI的抑制剂同时也是P450酶的底物，当酶活性被抑制后，其自身代谢也会受到阻碍，造成在体内暴露量非平稳的增加；此外，反应性代谢物对P450酶的共价修饰可能会导致半抗原的产生，可能会引起自体免疫反应，造成严重后果(谢珊珊等.细胞色素P450酶的时间依赖性抑制研究及其在新药研发中的作用[J].中国新药与临床杂志.Chin J New Drugs Clin Rem，2013,32(6)：419-424)。
目前PRMT5抑制剂进入临床的药物有GSK-3326595、JNJ-64619178、PF-06939999等，仍需开发更多安全的PRMT5激酶抑制剂。
发明内容
本发明提供式(I')所示的化合物或其药学上可接受的盐，
Figure PCTCN2022106401-appb-000001
其中，R 1选自H或NR 1aR 1b；
R 1a和R 1b各自独立地选自H或C 1-4烷基；所述的C 1-4烷基可任选被一个或多个R 1c基团所取代，所述R 1c基团为C 3-6环烷基；
Y选自N或CR 2；
R 2选自H、卤素、卤素取代的C 1-4烷基、CN或-SO 2-C 1-4烷基；
L 1选自化学键、-CH 2-、-X-CH 2、-(CH 2) n-CH＝CH-、-CH＝CH-(CH 2) n-、C 3-6环烷基或5-6元杂芳基；
X选自O、S、NH或CH 2；
环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基；
R 3选自羟基或C 1-4烷基；
m选自0、1、2或者3；
n选自0或1；
环B选自
Figure PCTCN2022106401-appb-000002
R 4选自NR 4aR 4b或C 1-4烷基；
R 4a和R 4b各自独立地选自H或C 1-4烷基；
Figure PCTCN2022106401-appb-000003
代表单键或双键；
当
Figure PCTCN2022106401-appb-000004
为单键时，R 5和R 6各自独立地选自H、氘、卤素或C 1-4烷基；R 7和R 8各自独立地为H；
当
Figure PCTCN2022106401-appb-000005
为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或C 5-6环烷基。
在本发明的一些方案中，本发明提供式(I')所示的化合物或其药学上可接受的盐，其中式(I')所示的化合物或其药学上可接受的盐为式(I)所示的化合物或其药学上可接受的盐，
Figure PCTCN2022106401-appb-000006
其中，R 1选自H或NR 1aR 1b；
R 1a和R 1b各自独立地选自H或C 1-4烷基；所述的C 1-4烷基可任选被一个或多个R 1c基团所取代，R 1c基团为C 3-6环烷基；
R 2选自H、卤素、卤素取代的C 1-4烷基、CN或-SO 2-C 1-4烷基；
R 3选自羟基或C 1-4烷基；
R 4选自羟基、
Figure PCTCN2022106401-appb-000007
NR 4aR 4b或C 1-4烷基，优选地，R 4选自NR 4aR 4b或C 1-4烷基；
R 4a和R 4b各自独立地选自H或C 1-4烷基；
L 1选自化学键、-CH 2-、-X-CH 2、-(CH 2) n-CH＝CH-、-CH＝CH-(CH 2) n-、C 3-6环烷基或5-6元杂芳基；
X选自O、S、NH或CH 2；
环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基；
m选自0、1、2或者3；
n选自0或1；
Figure PCTCN2022106401-appb-000008
代表单键或双键；
当
Figure PCTCN2022106401-appb-000009
为单键时，R 5和R 6各自独立地选自H、氘、卤素或C 1-4烷基；R 7和R 8各自独立 地为H；
当
Figure PCTCN2022106401-appb-000010
为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或C 5-6环烷基。
在本发明的一些方案中，式(I)所示的化合物或其药学上可接受的盐，
Figure PCTCN2022106401-appb-000011
其中，R 1选自H或NR 1aR 1b；
R 1a和R 1b各自独立地选自H或C 1-4烷基；所述的C 1-4烷基可任选被一个或多个R 1c基团所取代，R 1c基团为C 3-6环烷基；
R 2选自H或卤素；
R 3选自羟基或C 1-4烷基；
R 4选自羟基、
Figure PCTCN2022106401-appb-000012
NR 4aR 4b或C 1-4烷基，优选地，R 4选自NR 4aR 4b或C 1-4烷基；
R 4a和R 4b各自独立地选自H或C 1-4烷基；
L 1选自化学键、-CH 2-、-X-CH 2、-(CH 2) n-CH＝CH-、C 3-6环烷基或5-6元杂芳基；
X选自O、S、NH或CH 2；
环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基；
m选自0、1、2或者3；
n选自0或1；
Figure PCTCN2022106401-appb-000013
代表单键或双键；
当
Figure PCTCN2022106401-appb-000014
为单键时，R 5和R 6各自独立地选自H、氘或卤素；R 7和R 8各自独立地为H；
当
Figure PCTCN2022106401-appb-000015
为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或C 5-6环烷基。
在本发明的一些方案中，所述R 1选自H、氨基或
Figure PCTCN2022106401-appb-000016
在本发明的一些方案中，所述R 2选自H、F、Cl或Br。
在本发明的一些方案中，所述R 2选自卤素取代的C 1-4烷基、CN或-SO 2-C 1-4烷基。
在本发明的一些方案中，所述R 2选自CF 3、CN或-SO 2-CH 3。
在本发明的一些方案中，所述L 1选自化学键、-CH 2-、-O-CH 2-、-NH-CH 2-、-CH 2-CH 2-、-CH＝CH-、-CH 2-CH＝CH-、
Figure PCTCN2022106401-appb-000017
在本发明的一些方案中，所述L 1选自-CH＝CH-CH 2-。
在本发明的一些方案中，所述环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基，所述的杂环基含有1个或多个选自N或者O的杂原子。
本发明某些实施方案，环A选自C 5-6环烷基、C 5-6环烯基、5-10元单杂环、5-10元螺杂环、5-10元桥杂环或5-10元稠杂环。
本发明某些实施方案，环A选自C 5-6环烷基、C 5-6环烯基、5-6元单杂环、9-10元螺杂环、7-10元桥杂环或8-10元稠杂环。
在本发明的一些方案中，所述环A选自
Figure PCTCN2022106401-appb-000018
Figure PCTCN2022106401-appb-000019
在本发明的一些方案中，所述R 3选自羟基或甲基；m选自0、1、2或者3。
在本发明的一些方案中，所述结构单元
Figure PCTCN2022106401-appb-000020
选自
Figure PCTCN2022106401-appb-000021
Figure PCTCN2022106401-appb-000022
本发明的一些方案中，所述结构单元
Figure PCTCN2022106401-appb-000023
选自
Figure PCTCN2022106401-appb-000024
Figure PCTCN2022106401-appb-000025
在本发明的一些方案中，所述R 4选自氨基、甲基或-NHCH 3。
在本发明的一些方案中，所述
Figure PCTCN2022106401-appb-000026
代表单键或双键；
当
Figure PCTCN2022106401-appb-000027
为单键时，R 5和R 6各自独立地选自H、氘、F或甲基；R 7和R 8各自独立地为H；
当
Figure PCTCN2022106401-appb-000028
为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或环戊基。
在本发明的一些方案中，所述结构单元
Figure PCTCN2022106401-appb-000029
选自
Figure PCTCN2022106401-appb-000030
Figure PCTCN2022106401-appb-000031
在本发明的一些方案中，所述结构单元
Figure PCTCN2022106401-appb-000032
选自
Figure PCTCN2022106401-appb-000033
在本发明的一些方案中，所述结构单元
Figure PCTCN2022106401-appb-000034
为
Figure PCTCN2022106401-appb-000035
在本发明的一些方案中，所述化合物或其药学上可接受的盐，其中所述的化合物具有如下式(II)所示的结构：
Figure PCTCN2022106401-appb-000036
其中，
R 9选自H或C 1-4烷基；优选地，R 9选自H或甲基；
R 1、R 2、R 3、R 4、R 5、R 6、R 7、R 8、L 1和m如式(I’)或式(I)所定义。
在本发明某些实施方案中，式(II)化合物或其药效上可接受的盐中，R 9选自H或C 1-4烷基；优选地，R 9选自H或甲基；m选自1或2；R 1、R 2、R 3、R 4、R 5、R 6、R 7、R 8和L 1如式(I’)或式(I)所定义。
在本发明的一些方案中，上述化合物、或其药学上可接受的盐，其选自：
Figure PCTCN2022106401-appb-000037
其中R 1、R 2、R 4、R 5、R 6、R 7、R 8和R 9如式(II)所定义。
在本发明的一些方案中，上述化合物、其异构体或药学上可接受的盐，其选自：
Figure PCTCN2022106401-appb-000038
其中R 1、R 2、R 4和R 9如式(II)所定义。
本发明还提供下述化合物、其异构体或药学上可接受的盐，其选自：
Figure PCTCN2022106401-appb-000039
Figure PCTCN2022106401-appb-000040
Figure PCTCN2022106401-appb-000041
Figure PCTCN2022106401-appb-000042
本发明还涉及药物组合物，其包含所述化合物、或其药学上可接受的盐以及药学上可接受的载体、稀释剂或赋形剂。
本发明还涉及所述化合物或其药学上可接受的盐或其药物组合物在制备治疗癌症药物中的应用。
本发明还涉及所述化合物或其药学上可接受的盐，或其药物组合物在治疗癌症中的用途。
本发明还涉及一种治疗癌症的方法，其包括向有需要的对象施以有效剂量的所述化合物或其药学上可接受的盐，或其药物组合物。
本发明化合物对PRMT5甲基化酶以及人胰腺癌细胞株MIA PaCa-2具有较好的抑制作用，同时具有心脏安全性更高的潜在优势，对CYP3A4酶的抑制作用小，发生药物相互作用(DDI)的可能性更低，具有显著时间依赖性抑制(time-dependentinhibition，TDI)的风险较低，且具有良好的药代动力学性质。与现有技术相比，本发明化合物安全性更高。
术语解释
除非另有说明，本文所用的下列术语和短语旨在具有下列含义。一个特定的术语或短语在没有特别定义的情况下不应该被认为是不确定的或不清楚的，而应该按照普通的含义去理解。
术语“药学上可接受的”指在合理的医学判断范围内适合与人类和动物的组织接触使用而无过度的毒性、刺激、过敏反应或其它的问题或并发症，与合理的收益/风险比相当的那些化合物、材料、组合物和/或剂型。
术语“药学上可接受的盐”是指本发明化合物与相对无毒的酸或碱制备得到的衍生物。这些盐可以在化合物合成、分离、纯化期间就被制备，或者单独使用经过纯化的化合物的游离形式与适合的酸或碱反应。当化合物中含有相对酸性的官能团时，与碱金属、碱土金属氢氧化物或有机胺反应得到碱加成盐，包括基于碱金属与碱土金属的阳离子。当化合物中含有相对碱性的官能团时，与有机酸或无机酸反应得到酸加成盐。
本发明的化合物存在异构体，例如顺反异构体、对映异构体、非对映异构体、及其外消旋混合物和其他混合物，所有这些混合物都属于本发明的范围之内。
术语“对映异构体”是指互为镜像关系的立体异构体。
术语“非对映异构体”是指分子具有两个或多个手性中心，并且分子间为非镜像的关系的立体异构体。
术语“顺反异构体”是指分子中双键或者成环碳原子单键不能自由旋转而存在的构型。
除非另有说明，用楔形实线键
Figure PCTCN2022106401-appb-000043
和楔形虚线键
Figure PCTCN2022106401-appb-000044
表示一个立体中心的绝对构型。
本发明化合物的立体异构体可以通过手性合成或手性试剂或者其他常规技术制备。例如本发明某化合物的一种对映体，可以通过不对称催化技术或者手性助剂衍生技术制备得到。或者通过手性拆分技术，从混合物中得到单一立体构型的化合物。或者用手性起始原料，直接制备得到。本发明中的光学纯化合物的分离通常是使用制备色谱完成的，采用手性色谱柱，达到分离手性化合物的目的。
本发明中手性碳原子用*标记。例如
Figure PCTCN2022106401-appb-000045
中环戊基上的碳原子为手性碳，包含
Figure PCTCN2022106401-appb-000046
两个结构的化合物。
本发明中通过手性色谱柱分离得到的异构体用P1和P2表示，例如实施例6-P1和实施例6-P2代表实施例6化合物通过手性拆分得到的两种异构体。
本发明还包括同位素标记的化合物，包括氢、碳、氮、氧、磷、硫、氟和氯的同位素，分别例如 2H、 3H、 13C、 11C、 14C、 15N、 18O、 17O、 31P、 32P、 35S、 18F和 36Cl。含有上述 同位素和/或其它原子的其它同位素的本发明化合物都属于本发明的范围。优选地，同位素选自下组： 2H、 3H、 11C和 18F。更优选地，放射性同位素是 2H。具体地说，氘代的化合物旨在包含在本发明的范围内。
当一个连接基团的数量为0时，比如-(CH 2) n-CH＝CH-，n＝0表示该连接基团为化学键，即-CH＝CH-。
当一个取代基的键可以交叉连接到一个环上时，这种取代基可以与这个环上的任意原子相键合。例如，结构单元
Figure PCTCN2022106401-appb-000047
表示取代基R 3可以在环A上的任意一个位置发生取代。
术语“药学上可接受的载体”是指本领域通常可接受的用于将生物活性药剂递送给动物、特别是哺乳动物的介质，根据给药方式和剂型的性质包括佐剂、赋形剂或赋形物，例如稀释剂、防腐剂、填充剂、流动调节剂、崩解剂、润湿剂、乳化剂、助悬剂、甜味剂、调味剂、芳香剂、抗菌剂、抗真菌剂、润滑剂和分散剂。药学上可接受的载体在本领域普通技术人员的眼界范围内根据大量因素配制。其包括但不限于：配制的活性药剂的类型和性质，要将含有该药剂的组合物给药的对象，组合物的预期给药途径，和目标治疗适应症。药学上可接受的载体包括含水介质和非水介质这两者以及多种固体和半固体剂型。除了活性药剂以外，这样的载体包括许多不同的成分和添加剂，因多种原因(例如稳定活性药剂、粘合剂等)在处方中包括的另外的成分对于本领域普通技术人员是众所周知的。术语“赋形剂”通常是指配制有效的药物组合物所需要载体、稀释剂和/或介质。
术语“有效预防或治疗量”是指本发明化合物或其药学上可接受的盐指以适用于任何医学治疗和/或预防的合理效果/风险比治疗障碍的足够量的化合物。但应认识到，本发明式(I)所示化合物或其药学上可接受的盐和组合物的总日用量须由主诊医师在可靠的医学判断范围内作出决定。对于任何具体的患者，具体的治疗有效剂量水平须根据多种因素而定，所述因素包括所治疗的障碍和该障碍的严重程度；所采用的具体化合物的活性；所采用的具体组合物；患者的年龄、体重、一般健康状况、性别和饮食；所采用的具体化合物的给药时间、给药途径和排泄率；治疗持续时间；与所采用的具体化合物组合使用或同时使用的药物；及医疗领域公知的类似因素。例如，本领域的做法是，化合物的剂量从低于为得到所需治疗效果而要求的水平开始，逐渐增加剂量，直到得到所需的效果。
术语“任选被取代”是指可以被取代，也可以不被取代，除非另有规定，取代基的种类和数目在化学上可以实现的基础上可以是任意的，例如，术语“可任选地被一个或多个R 1c所取代”是指可以被一个或多个R 1c取代，也可以不被R 1c取代。当任何变量(例如R 1c)在化合物的组成或结构中出现一次以上时，其在每一种情况下的定义都是独立的。例如，如果一个基团被0-2个R 1c所取代，则所述基团可以任选地至多被两个R 1c所取代，并且每种情况下的R b都有独立的选项。
除非另有规定，“环”是指饱和的、部分饱和的或不饱和的单环以及多环。
除非另有规定，术语“杂环基”是指取代或未取代的饱和或不饱和的非芳香环，且包含1-3个选自N、O或S的杂原子。本发明所述的“杂环基”是指含有至少一个杂原子作为环原子、去除一个氢原子所衍生的、非芳族的环状基团；包括饱和或部分饱和的单环杂环基和多环杂环基；所述杂环基与连接位置无关(即，可以通过碳原子或杂原子结合)。所述多环杂环基包括稠杂环基、螺杂环基、桥杂环基。
所述的“稠杂环基”是指两个或两个以上的环相互间共用两个相邻的环原子所形成的环状结构，且其中至少一个环为杂环；所述稠杂环基包括单杂环基与单杂环基或环烷基或芳基或杂芳基稠和而成的环状结构，亦包括杂芳基与环烷基或单杂环基稠和而成的环状结构。
所述的“螺杂环基”是指两个或两个以上的环相互间共用一个环原子所形成的环状结构、且至少一个环为杂环。
所述的“桥杂环基”是指两个或两个以上的环相互间共用非相邻的环原子所形成的环状结构、且至少一个环为杂环。
本发明所述的杂环基优选为5-10元单杂环基、5-10元稠杂环基、5-10元螺杂环基或5-10元桥杂环基；进一步地，杂环基优选为5-6元单杂环基、8-10元稠杂环基、8-10元螺杂环基或7-9元桥杂环基。
“5-10元杂环基”实例包括，但不限于四氢吡咯基、哌啶基、哌嗪基、吗啉基、四氢呋喃基、四氢噻吩基、四氢吡喃基、1,3-二氧戊环、1,4-二氧六环、
Figure PCTCN2022106401-appb-000048
Figure PCTCN2022106401-appb-000049
等。
本发明中，当杂环基两端都被取代时为亚杂环基，例如结构单元
Figure PCTCN2022106401-appb-000050
环A选自5-10元杂环基时，其实质代表环A为5-10元亚杂环基，包括不限于
Figure PCTCN2022106401-appb-000051
Figure PCTCN2022106401-appb-000052
除非另有规定，术语“芳基”是指不饱和的、通常为芳族的烃基，其可为单环或稠合在一起的多个环。芳基的实例包括但不限于苯基、萘基。
除非另有规定，术语“杂芳基”意指稳定的单环或者多环的芳族烃，优选包含碳原子和1、2、3或4个独立地选自N、O和S的环杂原子。优选5-12元杂芳基，更优选5-6元杂芳基；杂芳基的实例包括但不限于吡咯基、吡唑基、咪唑基、吡嗪基、异恶唑基、噻唑基、 呋喃基、噻吩基、嘧啶基、吡啶基、
Figure PCTCN2022106401-appb-000053
本发明中，当杂芳基两端都被取代时为亚杂芳基，例如式(I’)或式(I)化合物中的L 1选自5-6元杂芳基时，其实质代表L 1为5-6元亚杂芳基，包括不限于
Figure PCTCN2022106401-appb-000054
Figure PCTCN2022106401-appb-000055
除非另有规定，“环烷基”是指饱和的单环或多环烃基。环烷基优选为C 3-8环烷基，更优选为C 3-6环烷基，进一步优选为C 5-6环烷基，环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己基、环庚基、环辛基或
Figure PCTCN2022106401-appb-000056
本发明中，当环烷基两端都被取代时为亚环烷基，例如结构单元
Figure PCTCN2022106401-appb-000057
中，环A选自C 5-6环烷基时，其实质代表环A为C 5-6亚环烷基，包括不限于
Figure PCTCN2022106401-appb-000058
除非另有规定，“环烯基”是指含有双键的单环或多环烃基。环烯基优选为C 3-6环烯基，更优选C 5-6环烯基，实例包括但不限于环丙烯、环丁烯、环戊烯、环己烯、1,3-环己二烯或1,4-环己二烯。
本发明中，当环烯基两端都被取代时为亚环烯基，例如结构单元
Figure PCTCN2022106401-appb-000059
环A选自C 5-6环烯基时，其实质代表环A为C 5-6亚环烯基，包括不限于
Figure PCTCN2022106401-appb-000060
本发明中环B选自
Figure PCTCN2022106401-appb-000061
的情形，当
Figure PCTCN2022106401-appb-000062
为双键时，R 5和R 8不存在，R 6和R 7 与其相连的C原子一起形成苯基或C 5-6环烷基，其所对应的
Figure PCTCN2022106401-appb-000063
是指
Figure PCTCN2022106401-appb-000064
除非另有规定，术语“烷基”用于表示直链或支链的饱和烃基。优选为C 1-6的烷基，更优选为C 1-4的烷基，烷基的实例包括，但不限于甲基，乙基，丙基、异丙基、丁基、异丁基、叔丁基、戊基、异戊基，新戊基、正己基等。
除非另有规定，术语“卤素”表示氟、氯、溴或碘原子。
特别说明，本文中所有的取代基和/或其变体的组合只有在这样的组合会产生稳定的化合物的情况下才是被允许的。
在本发明实施例中，标题化合物的命名是借助Chemdraw通过化合物结构转化过来的。若化合物名称与化合物结构存在不一致的情况，可通过综合相关信息和反应路线辅助确定；无法通过其他来确认的，以给出的化合物结构式为准。本发明中部分化合物的制备方法引用了前述类似化合物的制备方法。本领域人员应当知晓，在使用或参照使用其引用的制备方法时，反应物的投料比、反应溶剂、反应温度等可根据反应物的不同，进行适当的调整。
本发明的化合物可以通过本领域技术人员所熟知的多种合成方法来制备，包括下面列举的具体实施方式、其与其他化学合成方法的结合所形成的实施方式以及本领域技术上人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。
具体实施方式
下面通过实施例对本发明进行详细描述，但并不意味着对本发明任何不利限制。本文已经详细地描述了本发明，其中也公开了其具体实施例方式，对本领域的技术人员而言，在不脱离本发明精神和范围的情况下针对本发明具体实施方式进行各种变化和改进将是显而易见的。
实验仪器汇总：
本发明的化合物结构是通过核磁共振(NMR)或/和液质联用色谱(LC-MS)，或超高效液质联用色谱(UPLC-MS)来确定的。NMR化学位移(δ)以百万分之一(ppm)的单位给出。NMR的测定是用Bruker Neo 400M或者Bruker Ascend 400核磁仪器，测定溶剂为氘代二甲基亚砜(DMSO-d 6)，氘代甲醇(CD 3OD)和氘代氯仿(CDCl 3)，重水(D 2O)，内标为四甲基硅烷(TMS)。
液质联用色谱LC-MS的测定用Agilent 1260-6125B single quadrupole mass spectrometer，柱子为Welch Biomate column(C18,2.7μm,4.6×50mm)或者waters H-Class  SQD2，柱子为Welch Ultimate column(XB-C18,1.8μm,2.1×50mm)，质谱仪离子源为电喷雾离子化。
超高效液质联用色谱UPLC-MS的测定用Waters UPLC H-class SQD质谱仪(离子源为电喷雾离子化)。
HPLC的测定使用Waters e2695-2998或Waters ARC和Agilent 1260或Agilent Poroshell HPH高效液相色谱。
制备HPLC使用Waters 2555-2489(10μm，ODS 250cm×5cm)或GILSON Trilution LC，柱子为Welch XB-C18柱(5μm,21.2×150mm)。
手性HPLC测定使用waters acquity UPC2；柱子为Daicel chiralpak AD-H(5μm,4.6×250mm),Daicel chiralpak OD-H(5μm,4.6×250mm),Daicel chiralpak IG-3(3μm,4.6×150mm),Chiral Technologies Europe AD-3(3μm,3.0×150mm)和Trefoil TM Technology Trefoil TM AMY1(2.5μm,3.0×150mm)。
超临界流体色谱(SFC)使用waters SFC 80Q，柱子为Daicel Chiralcel OD/OJ/OZ(20×250mm,10μm)或Daicel Chiralpak IC/IG/IH/AD/AS(20×250mm,10μm)。
薄层层析硅胶板使用烟台江友硅胶开发有限公司GF254硅胶板或乳山市上邦新材料有限公司GF254硅胶板，TLC采用的规格是0.15mm～0.20mm，制备型20×20cm，柱层析一般使用于成化工200～300目硅胶为载体。
本发明实施例中的起始原料是已知的并且可以在市场上买到，或者可以采用或按照本领域已知的方法来合成。
在无特殊说明的情况下，本发明的所有反应均在连续的磁力搅拌下，在干燥氮气或氩气气氛下进行，溶剂为干燥溶剂，反应温度单位为摄氏度或℃。
本申请部分制备实施例获得了化合物的三氟乙酸盐，本领域技术人员可以理解，通过三氟乙酸盐制备化合物，或通过化合物制备三氟乙酸盐均为较常规的手段，本申请所公开的化合物三氟乙酸盐结构或其制备可以视为等同公开了游离体化合物结构及其制备。
本发明实施例中使用的缩写及其对应的化学名称如下：
Figure PCTCN2022106401-appb-000065
Figure PCTCN2022106401-appb-000066
中间体：Int-1
3-溴-7-碘喹啉-2-胺
Figure PCTCN2022106401-appb-000067
反应路线:
Figure PCTCN2022106401-appb-000068
操作步骤：
步骤A:向反应瓶中加入7-硝基喹啉(25g，143mmol)和醋酸溶液(50mL)。置换反应体系为氮气体系，然后冷却到0℃。向反应体系中逐滴滴加N-溴代琥珀酰亚胺(50.6g，284mmol)，反应体系继续在100℃搅拌2小时。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(500mL×3次)萃取，合并有机相，有机相用饱和食盐水(300mL×3次)洗涤，无水硫酸钠干燥，过滤，滤液减压浓缩得到3-溴-7-硝基喹啉(24.5g)。
MS(ESI)M/Z:253.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ9.18(d,J＝2.0Hz,1H),8.97(s,1H),8.79(t,J＝11.4Hz,1H),8.41(dd,J＝9.0,2.0Hz,1H),8.23(d,J＝9.0Hz,1H).
步骤B:向反应瓶中加入3-溴-7-硝基喹啉(24.5g，97mmol)和氯化铵(20.0g，374mmol)溶于混合溶剂(乙醇/水＝1.5/1，体积比，250mL)中，在室温中缓慢加入锌粉(65.4 g，1000mmol)，然后体系置换为氮气氛围。滴加完毕后，反应体系在80℃下搅拌2小时。
TLC监测显示原料消失后，向反应液中缓慢加入水淬灭。然后加入二氯甲烷(50mL)和甲醇(5mL)，搅拌30分钟，过滤反应液。滤液浓缩得到油状粗产物，用硅胶柱层析纯化得到3-溴喹啉-7-胺(6.2g)。
MS(ESI)M/Z:223.0[M+H] +.
步骤C：向反应瓶中加入硫酸(108mL)，然后0℃下缓慢加入3-溴喹啉-7-胺(6.2g，27.8mmol)溶液(86mL)。在0℃下搅拌10分钟之后，将亚硝酸钠(3.8g，55.1mmol)溶液(6mL)缓慢滴入。随后继续搅拌30分钟，在0℃下向反应体系中缓慢加入碘化钠(12.5g，83.4mmol)溶液。反应液在60℃下搅拌2小时。
LCMS监测显示原料消失后，将反应液冷却到室温，向反应液中加入乙酸乙酯(50mL)搅拌10分钟，然后过滤，滤饼反复用二氯甲烷和甲醇的混合溶液洗涤，合并滤液，减压浓缩。所得残余物用硅胶柱层析纯化得到3-溴-7-碘喹啉(4g，收率25.6％)。
MS(ESI)M/Z:333.8[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ9.16(dd,J＝5.3,2.3Hz,1H),8.96(t,J＝3.6Hz,1H),8.82(dd,J＝7.8,2.2Hz,1H),8.40(dd,J＝9.0,2.3Hz,1H),8.23(d,J＝9.0Hz,1H).
步骤D：在室温下，将3-溴-7-碘喹啉(500mg，1.5mmol)溶于二氯甲烷(10mL)中。在0℃下缓慢加入间氯过苯甲酸(1.03g，6mmol)，反应液在室温下搅拌12小时。
LCMS监测反应，0℃下向反应液中加入水(30mL)淬灭，然后缓慢加入氯化铵的饱和水溶液调节pH至7，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，收集滤液减压浓缩。所得残余物用硅胶柱层析纯化得到3-溴-7-碘喹啉1-氧化物(160mg)。
MS(ESI)M/Z:349.8[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.96(d,J＝2.3Hz,1H),8.73(t,J＝13.1Hz,1H),8.44(d,J＝14.5Hz,1H),7.97(dd,J＝8.6,1.5Hz,1H),7.77(d,J＝8.6Hz,1H).
步骤E：在室温下，将3-溴-7-碘喹啉1-氧化物(160mg，0.45mmol)，溶于三氯甲烷(10mL)中。然后在0℃下缓慢加入三氯氧磷(10mL)，反应体系抽真空换氮气多次。反应液在70℃油浴下搅拌12小时。
LCMS监测显示原料消失后，减压浓缩。加入二氯甲烷(20mL)然后用饱和碳酸氢钠溶液调节pH值到7，混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到3-溴-2-氯-7-碘喹啉(90mg)。
MS(ESI)M/Z:367.8[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.96(s,1H),8.44(d,J＝18.2Hz,1H),8.03(t,J＝18.1Hz,1H),7.79(t,J＝10.1Hz,1H).
步骤F：在室温下，将3-溴-2-氯-7-碘喹啉(90mg，0.24mmol)溶于混合溶液(乙醇/氨水＝1/1，体积比，8mL)中。随后反应液在微波100℃下搅拌20小时。
LCMS监测显示原料消失后，减压浓缩，混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到3-溴-7-碘喹啉-2-胺(76mg)。
MS(ESI)M/Z:348.8[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.39(d,J＝10.2Hz,1H),7.86(s,1H),7.48(dt,J＝19.4,5.0Hz,2H),6.84(brs,2H).
实施例1
(1R,2S,3R,5S)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000069
反应路线:
Figure PCTCN2022106401-appb-000070
Figure PCTCN2022106401-appb-000071
操作步骤：
步骤A：将6-肼基嘧啶-4-醇(2.0g，15.9mmol)和环戊酮(1.3g，15.5mmol)溶于乙醇(15mL)。反应体系加热至80℃并搅拌3小时。
LCMS监测显示原料消失后，将反应液减压并将所得固体溶于二苯醚(20mL)。反应体系加热至250℃并搅拌8小时。待反应液冷却至室温，向反应液中加入甲基叔丁基醚(20mL)，反应液中产生大量固体，过滤并用甲基叔丁基醚(10mL)洗涤固体，滤饼用减压蒸馏悬干得到5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-酚粗产品(2.7g)。
MS(ESI)M/Z:176.2[M+H] +.
步骤B：将5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-酚(2.7g，15.4mmol)溶于三氯氧磷(20mL)。反应体系加热至100℃并搅拌3小时。LCMS监测显示原料消失后，在减压蒸馏下去除大部分三氯氧磷。反应液用乙酸乙酯(50mL)稀释并缓慢滴入氢氧化钠(1N，50mL)水溶液中，萃取有机相，有机相用饱和食盐水(300mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到4-氯-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(370mg)。
1H NMR(400MHz,DMSO-d 6)δ12.35(s,1H),8.44(s,1H),2.94-2.84(m,4H),2.49-2.42(m,2H).
步骤C：在室温条件下，将4-氯-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(243.0mg，1.3mmol)溶于四氢呋喃(4mL)中。随后，向其中加入叔丁醇钾(141.3mg，1.3mmol)。反应液在室温下继续搅拌2小时。减压蒸馏除去溶剂，加入二异丙基乙氨(2mL)并超声5分钟，过滤，收集滤饼，减压浓缩得化合物4-氯-6,7-二氢-5H-环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8-化钾(291.1mg，收率96.9％)。
步骤D：在一个单口瓶中，加入三氟甲磺酸酐(1.16g，4.11mmol)的二氯甲烷(20mL)溶液，冰浴下向反应液中滴加(3aS,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(500mg，2.74mmol，参考文献J.Org.Chem.2004,69,3993-3996方法制备，其全文内容通过引用结合到本申请中)和吡啶(649mg，8.22mmol)的二氯甲烷混合液(10mL)。反应液在此温度下继续搅拌半小时。
TLC监测显示原料消失后，向反应液中加入冰水淬灭。反应液用二氯甲烷(20mL×3次)萃取，合并有机相，有机相用饱和食盐水(10mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲烷磺酸酯(685mg)。
1H NMR(400MHz,CDCl 3)δ5.83-5.69(m,1H),5.19-5.06(m,2H),5.05-4.96(m,1H),4.63(dd,J＝5.5Hz,1H),4.51(dd,J＝5.9,1.8Hz,1H),2.94-2.81(m,1H),2.45-2.31(m,1H),2.13-2.02(m,1H),1.53(s,3H),1.34(s,3H).
步骤E：在0℃下，将4-氯-6,7-二氢-5H-环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8-化钾(291.1mg，1.26mmol)溶于N,N-二甲基甲酰胺(4mL)中，然后将(3aR,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲烷磺酸酯(331.8mg，1.05mmol)溶于N,N-二甲基甲酰胺(0.5mL)缓慢加入反应液中。反应液在室温下继续搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物4-氯-8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(300mg)。
MS(ESI)M/Z:360.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.52(s,1H),5.96(m,1H),5.25-5.06(m,2H),5.06-4.98(m,1H),4.92(dd,J＝7.3,5.4Hz,1H),4.56(t,J＝7.0Hz,1H),3.20-3.06(m,1H),3.06-2.95(m,1H),2.95-2.86(m,2H),2.79-2.68(m,1H),2.46-2.51(m,2H)2.38-2.24(m,2H),1.49(s,3H),1.22(s,3H).
步骤F：在室温下，将4-氯-8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(180mg，0.50mmol)，7-溴喹啉-2-胺(133.5mg，0.60mmol)和四乙基氯化铵(91.1mg，0.55mmol)溶于N,N-二甲基甲酰胺(5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(22.4mg，0.1mmol)，N,N-二异丙基乙胺(322.5mg，2.5mmol)。反应液在80℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(140mg)。
MS(ESI)M/Z:502.0[M+H] +.
步骤G：在室温条件下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(45mg，0.09mmol)溶于氨水/异丙醇(2mL/0.8mL)中，反应体系封管加热至120℃并搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到化合物8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢基-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-4-胺(21.0mg)。
MS(ESI)M/Z:483.2[M+H] +.
步骤H：将8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢基-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-4-胺(15mg，0.03mmol)和钯炭(5mg)溶于甲醇(4mL)，反应体系先用氮气置换空气，然后再用氢气置换两次，反应在室温和氢气条件下搅拌3小时。
LCMS监测显示原料消失后，将反应过滤去除钯炭后液缩得到粗产品化合物8-((3aS,4R,6S,6aR)-6-(2-(2-氨基喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(10.0mg，收率68.7％)。
MS(ESI)M/Z:485.2[M+H] +.
步骤I：在室温下，将8-((3aS,4R,6S,6aR)-6-(2-(2-氨基喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(10mg，0.02mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5S)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊烷-1,2-二醇(2.0mg)。
MS(ESI)M/Z:445.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.95(s,1H),7.82(d,J＝8.8Hz,1H),7.53(d,J＝8.1Hz,1H),7.27(s,1H),7.03(d,J＝8.1Hz,1H),6.68(d,J＝8.8Hz,1H),6.43(s,2H),6.32(s,2H),4.79-4.66(m,2H),4.62(d,J＝4.7Hz,1H),4.25(dd,J＝13.7,6.3Hz,1H),3.77-3.68(m,1H),2.85(d,J＝5.2Hz,4H),2.77-2.62(m,2H),2.41-2.33(m,2H),2.19-2.06(m,1H),1.99-1.79(m,2H),1.72-1.51(m,2H).
实施例2
(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8-(5H)-基)-5-((E)-2-(2-氨基喹啉-7-基)乙烯基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000072
反应路线:
Figure PCTCN2022106401-appb-000073
操作步骤：
步骤A：在室温下，8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(14mg，0.03mmol)溶于甲醇(1.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1.5mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-((E)-2-(2-氨基喹啉-7-基)乙烯基)环戊烷-1,2-二醇(4.0mg)。
MS(ESI)M/Z:443.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.98(s,1H),7.82(d,J＝8.8Hz,1H),7.56(d,J＝8.3Hz,1H),7.44-7.26(m,2H),6.68(d,J＝8.8Hz,1H),6.59-6.53(m,2H),6.46(s,2H),6.38(s,2H),4.86(t,J＝5.4Hz,2H),4.83-4.73(m,1H),4.29(dd,J＝12.5,6.2Hz,1H),3.93(q,J＝6.0Hz,1H),2.96-2.80(m,4H),2.77-2.64(m,1H),2.43-2.35(m,2H),2.25-2.11(m,1H),2.05-1.82(m,1H).
实施例3
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000074
反应路线:
Figure PCTCN2022106401-appb-000075
操作步骤：
步骤A：在室温条件下，将4-氯-8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(400mg，1.11mmol)溶于氨水/异丙醇(7mL/5mL)中，反应体系封管加热至120℃并搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到化合物8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(170.0mg，收率45.0％)。
MS(ESI)M/Z:341.0[M+H] +.
步骤B：在室温下，将8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(30mg，0.09mmol)，3-溴-7-碘喹啉-2-胺(37.1mg，0.11mmol)和四乙基氯化铵(16.0mg，0.10mmol)溶于N,N-二甲基甲酰胺(3mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸 钯(3.9mg，0.02mmol)，N,N-二异丙基乙胺(56.8mg，0.44mmol)。反应液在70℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(26mg)。
MS(ESI)M/Z:560.8[M+H] +.
步骤C：在室温下，将8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(40mg，0.07mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇(10.0mg)。
MS(ESI)M/Z:520.8[M+H] +.
1H NMR(400MHz,MeOD)δ8.24(s,1H),8.01(s,1H),7.56(d,J＝8.4Hz,1H),7.48(s,1H),7.44(d,J＝8.5Hz,1H),6.76-6.51(m,2H),4.94-4.89(m,1H),4.49(t,J＝6.4Hz,1H),4.12(t,J＝6.4Hz,1H),3.03-2.82(m,5H),2.64-2.46(m,2H),2.39-2.28(m,1H),2.05(dd,J＝23.3,10.9Hz,1H).
实施例4
(1S,2R,3R,5R)-3-((E)-2-(2-氨基喹啉-7-基)乙烯基)-5-(4-(甲胺基)-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000076
反应路线:
Figure PCTCN2022106401-appb-000077
操作步骤：
步骤A：在室温条件下，将4-氯-8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(200mg，0.56mmol)溶于甲氨/异丙醇(1mL/5mL)中，反应体系封管加热至120℃并搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-N-甲基-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(140.0mg)。
MS(ESI)M/Z:355.0[M+H] +.
步骤B：在室温下，将8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-N-甲基-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(100mg，0.28mmol)，7-溴喹啉-2-胺(75.2mg，0.34mmol)和四乙基氯化铵(51.0mg,0.31mmol)溶于N,N-二甲基甲酰胺(5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(12.6mg，0.056mmol)，N,N-二异丙基乙胺(180.6mg，1.4mmol)。反应液在100℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(50mL)淬灭，混合液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-N-甲基-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(80mg)。
MS(ESI)M/Z:497.0[M+H] +.
步骤C：在室温下，将8-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-N-甲基-5,6,7,8-四氢环戊二烯并[4,5]吡咯并 [2,3-d]嘧啶-4-胺(40mg，0.07mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基喹啉-7-基)乙烯基)-5-(4-(甲胺基)-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇(35.0mg)。
MS(ESI)M/Z:457.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.28(s,1H),7.11(t,J＝9.0Hz,1H),6.85-6.75(m,1H),6.68(s,1H),6.66-6.54(m,1H),5.98(dd,J＝8.8,4.1Hz,1H),5.88-1.87(m,1H),5.82-5.69(m,1H),4.18-4.10(m,1H),3.70(t,J＝6.5Hz,1H),3.34(t,J＝6.4Hz,1H),2.23(s,3H),2.20-2.01(m,5H),1.79-1.67(m,2H),1.59-1.50(m,1H),1.29-1.21(m,1H).
实施例5
(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000078
反应路线:
Figure PCTCN2022106401-appb-000079
操作步骤：
步骤A:向反应瓶中加入化合物8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(80mg，0.235mmol)的四氢呋喃(3mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(2.35mmol)的四氢呋喃溶液(0.5M，4.7mL)，滴加完毕后，反应液被加热到65℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(249.1mg，1.175mmol)的水溶液(1mL)。反应液在室温下搅拌30分钟后，加入3-溴-7-碘喹啉-2-胺(79mg，0.225mmol)和PdCl 2(dppf)(18mg，0.024mmol)的四氢呋喃(1mL) 溶液。最后所得的反应混合液被加热到65℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到8-((3aS,4R,6S,6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(60mg)。
MS(ESI)M/Z:562.8[M+H] +,564.8[M+H+2] +.
步骤B：在室温下，将化合物8-((3aS,4R,6S,6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(60mg，0.106mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于甲醇(2mL)，然后加入7M氨甲醇溶液调节pH值约等于8-9。所得溶液经制备型高效液相色谱纯化得到最终产物(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇(5.5mg)。
MS(ESI)M/Z:523.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.26(s,1H),7.97(s,1H),7.56(d,J＝8.2Hz,1H),7.41(s,1H),7.20(dd,J＝8.3,1.5Hz,1H),4.82-4.77(m,1H),4.44(dd,J＝7.9,6.2Hz,1H),3.94-3.86(m,1H),3.01-2.74(m,6H),2.56-2.46(m,2H),2.36-2.25(m,1H),2.12-1.96(m,2H),1.88-1.69(m,2H).
实施例6
(1S,2R,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-((1R)-2-(2-氨基喹啉-7-基)环丙基)环戊烷-1,2-二醇及其异构体
Figure PCTCN2022106401-appb-000080
反应路线:
Figure PCTCN2022106401-appb-000081
操作步骤：
步骤A：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(250.0mg，0.50mmol)，二碳酸二叔丁酯(326.8mg，1.5mmol)和4-二甲氨基吡啶(12.2mg，0.1mmol)溶于四氢呋喃(12mL)中。随后，向上述溶液中加入三乙胺(202.4mg，2.0mmol)。反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。向粗产品中加入水(100mL)淬灭，混合液用乙酸乙酯(100mL×3次)萃取，合并有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(250mg，收率71.2％)。
MS(ESI)M/Z:724.3[M+Na] +.
步骤B：在-10℃下，将氢氧化钾(1.7g，25mL水)水溶液缓慢加入到乙醚(60mL)中，再加入1-甲基-1-亚硝基脲(3.0g，29.1mmol)反应液在-10℃下继续搅拌1小时。将反应液冷却至-78℃，水相冻住倒出有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，得到现制备的重氮甲烷的乙醚溶液(50mL)。
在室温下，将二叔丁基(7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(150mg，0.21mmol)和醋酸钯(43.2mg，0.19mmol)溶于四氢呋喃(6mL)，将反应体系置换氮气并冷却到-78℃，将现制备的重氮甲烷的乙醚溶液(50mL)缓慢加入到反应液中。反应体系缓慢回到室温，并在在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-基)氨基甲酸酯(102mg)。
MS(ESI)M/Z:616.3[M+H-Boc] +.
步骤C：在室温条件下，将二叔丁基(7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-基)氨基甲酸酯(150.0mg，0.21mmol)溶于氨水/异丙醇(10mL/3mL)中，反应体系封管加热至120℃并搅拌24小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩得到粗产品8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-氨基喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(90mg)。
MS(ESI)M/Z:497.2[M+H] +.
步骤D：在室温下，将8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-氨基喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(90mg，0.19mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速：20毫升/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品，减压冻干。得到(1S,2R,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-((1R)-2-(2-氨基喹啉-7-基)环丙基)环戊烷-1,2-二醇的两个异构体(实施例6-P1：15.0mg，实施例6-P2：13.0mg)。
实施例6-P1：
HPLC：保留时间6.254分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:457.2[M+H] +.
1H NMR(400MHz,MeOD)δ7.91(s,1H),7.84(d,J＝8.8Hz,1H),7.50(d,J＝8.3Hz,1H),7.23(s,1H),7.00(dd,J＝8.3,1.6Hz,1H),6.71(d,J＝8.8Hz,1H),4.93-4.85(m,1H),4.56(dd,J＝8.4,5.7Hz,1H),4.08(dd,J＝5.6,4.2Hz,1H),2.95-2.70(m,4H),2.52-2.27(m,3H),2.02-1.83(m,2H),1.80-1.65(m,1H),1.51-1.29(m,1H),1.18-1.02(m,2H).
实施例6-P2：
HPLC：保留时间6.533分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈 0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
实施例7
(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-氨基喹啉-7-基)氧基)甲基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000082
反应路线:
Figure PCTCN2022106401-appb-000083
操作步骤：
步骤A:在一个单口瓶中，加入(3aR,6aR)-2,2-二甲基-3a,6a-二氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(830mg，5.39mmol)，二苯甲酮(196mg，1.08mmol)和甲醇(350mL)。然后用氮气连续鼓泡1小时，所得反应液在高压汞灯的照射下搅拌1小时(暗室操作)。
TLC监测显示原料消失后，反应液减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,6R,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(630mg)。
1H NMR(400MHz,DMSO-d 6)δ4.92(t,J＝4.6Hz,1H),4.63(d,J＝5.4Hz,1H),4.18(d,J＝5.4Hz,1H),3.64-3.52(m,1H),3.46-3.38(m,1H),2.63-2.52(m,2H),2.41-2.34(m,1H),1.31(s,3H),1.26(s,3H).
步骤B:在一个单口瓶中，加入(3aR,6R,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(450mg，2.42mmol)，叔丁基二苯基氯硅烷(666mg，2.42mmol)和无水二氯甲烷(20mL)。反应瓶置于冰浴中降温至零℃，然后分批加入咪唑(329mg，4.84mmol)。所得反应液被升到室温，继续搅拌3小时。
TLC监测显示原料消失后，向反应液中加入冰水淬灭。反应液用二氯甲烷(30mL×3次)萃取，合并有机相，有机相用饱和食盐水(20mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(550mg)。
1H NMR(400MHz,DMSO-d 6)δ7.63-7.55(m,4H),7.52-7.42(m,6H),4.73-4.60(m,1H),4.21(d,J＝5.4Hz,1H),3.78(dd,J＝10.1,2.9Hz,1H),3.59(dd,J＝10.1,3.6Hz,1H),2.80-2.61(m,1H),2.54-2.52(m,1H),2.49-2.45(m,1H),1.33(s,3H),1.27(s,3H),0.95(s,9H).
步骤C:在一个三口瓶中加入(3aR,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(640mg，1.5mmol)的四氢呋喃(15mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后逐滴滴加四氢铝锂(3.0mmol)的四氢呋喃溶液(2.5M，1.2mL)。滴加完毕后，反应体系继续在0℃下搅拌1小时。
TLC监测显示原料消失后，向反应液中缓慢加入十水合硫酸钠淬灭直至没有气泡冒出。然后加入四氢呋喃搅拌30分钟，过滤反应液。滤液被浓缩得到油状粗产物，用硅胶柱层析纯化得到(3aS,4S,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(490mg)。
步骤D:在一个单口瓶中，加入三氟甲磺酸酐(486mg，1.73mmol)的二氯甲烷(15mL)溶液,冰浴下向反应液中滴加(3aS,4S,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(490mg，1.15mmol)和吡啶(272mg，3.45mmol)的二氯甲烷的混合液(5mL)。反应液在此温度下继续搅拌半小时。
TLC监测显示原料消失后，向反应液中加入冰水淬灭。反应液用二氯甲烷(20mL×3次)萃取，合并有机相，有机相用饱和食盐水(10mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,4S,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲基磺酸酯(640mg)。
步骤E:冰浴下，向4-氯-6,7-二氢-5H-环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8-钾盐(266mg，1.15mmol)的N,N-二甲基甲酰胺(8mL)溶液逐滴滴加(3aR,4S,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲基磺酸酯 (640mg，1.15mmol)的N,N-二甲基甲酰胺(2mL)溶液。滴加完毕后，反应液在室温下搅拌过夜。
TLC检测显示新点生成，反应液被倒入冰水中。反应液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相用饱和食盐水(10mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-氯-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(492mg)。
MS(ESI)M/Z:602.2[M+H] +.
步骤F:在一个三口瓶中加入8-((3aS,4R,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-氯-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(492mg，0.82mmol)的四氢呋喃(10mL)溶液，然后体系置换为氮气氛围，逐滴滴加四丁基氟化铵(2.0mmol)的四氢呋喃溶液(1.0M，2.0mL)。滴加完毕后，反应体系继续在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)甲醇(220mg)。
MS(ESI)M/Z:364.0[M+H] +.
步骤G:在室温下，将((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)甲醇(170mg，0.47mmol)和2-氯喹啉-7-醇(126mg，0.71mmol)溶于甲苯(2mL)中。将2-(三丁基亚正膦基)乙腈(453mg，1.88mmol)溶于甲苯(0.5mL)中。随后，在室温下向反应体系中缓慢加入2-(三丁基亚正膦基)乙腈的甲苯溶液。反应液在油浴下搅拌4小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(20mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-8-((3aS,4R,6R,6aR)-6-(((2-氯喹啉-7-基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(190mg)。
MS(ESI)M/Z:525.0[M+H] +.
步骤H:在室温条件下，将4-氯-8-((3aS,4R,6R,6aR)-6-(((2-氯喹啉-7-基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(190mg，0.36mmol)溶于氨水/异丙醇(5mL/1.5mL)中，反应体系封管加热至120℃并搅拌24小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩，所得残余物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-6-(((2-氨基喹啉-7-基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并 [d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-4-胺(190mg)。
MS(ESI)M/Z:487.0[M+H] +.
步骤I:在室温下，将8-((3aS,4R,6R,6aR)-6-(((2-氨基喹啉-7-基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯[2,3-d]嘧啶-4-胺(58mg，0.12mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-氨基喹啉-7-基)氧基)甲基)环戊烷-1,2-二醇(12.8mg)。
MS(ESI)M/Z:447.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.95(s,1H),7.78(d,J＝8.8Hz,1H),7.52(d,J＝8.8Hz,1H),6.90(d,J＝2.4Hz,1H),6.81(dd,J＝8.8,2.4Hz,1H),6.58(d,J＝8.8Hz,1H),6.43(s,2H),6.31(s,2H),4.96-4.84(m,1H),4.80(dd,J＝9.9,5.5Hz,2H),4.35(dt,J＝8.8,6.2Hz,1H),4.19-4.05(m,2H),3.94(dd,J＝8.8,4.4Hz,1H),2.98-2.80(m,4H),2.44-2.34(m,3H),2.19(dt,J＝12.6,8.4Hz,1H),1.85-1.75(m,1H).
实施例8
(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-((环丙基甲基)氨基)喹啉-7-基)氧基)甲基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000084
制备方法参考实施例7，最后得到目标产物(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-((环丙基甲基)氨基)喹啉-7-基)氧基)甲基)环戊烷-1,2-二醇。
MS(ESI)M/Z:501.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.95(s,1H),7.72(d,J＝8.8Hz,1H),7.49(d,J＝8.6Hz,1H),7.02(t,J＝5.4Hz,1H),6.96(d,J＝2.4Hz,1H),6.77(dd,J＝8.7,2.4Hz,1H),6.61(d,J＝8.8Hz,1H),6.43(s,2H),4.90(dd,J＝19.6,8.8Hz,1H),4.79(dd,J＝10.2,5.6Hz,2H),4.35(dt,J＝8.8,6.2Hz,1H),4.13(p,J＝9.8Hz,2H),3.94(dd,J＝8.6,4.6Hz,1H),3.28-3.22(m,2H),2.97-2.81(m,4H),2.45-2.35(m,3H),2.18(dt,J＝13.0,8.6Hz,1H),1.79(dd,J＝21.0,11.0Hz,1H),1.18-1.02(m,1H),0.50-0.43(m,2H),0.29-0.21(m,2H).
实施例9
(1S,2R,3R,5R)-3-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-5-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)环戊基-1,2-二醇及其异构体
Figure PCTCN2022106401-appb-000085
反应路线:
Figure PCTCN2022106401-appb-000086
操作步骤：
步骤A：在室温下，将4-氯-8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(137.2mg，0.38 mmol)，3-溴-7-碘喹啉-2-胺(140.0mg，0.40mmol)和四乙基氯化铵(69.2mg，0.42mmol)溶于N,N-二甲基甲酰胺(6mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(17.1mg，0.07mmol)，N,N-二异丙基乙胺(245.1mg，1.9mmol)。反应液在70℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(100mL)淬灭，混合液用乙酸乙酯(100mL×3次)萃取，合并有机相，有机相先用饱和食盐水(150mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物3-溴-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)喹啉-2-胺(125mg)。
MS(ESI)M/Z:580.0[M+H] +.
步骤B：在室温下，将3-溴-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)喹啉-2-胺(145.0mg，0.25mmol)，二碳酸二叔丁酯(191.3mg，0.88mmol)和4-二甲氨基吡啶(6.1mg，0.05mmol)溶于四氢呋喃(7mL)中。随后，向上述溶液中加入三乙胺(126.5mg，1.25mmol)。反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。向粗产品中加入水(50mL)淬灭，混合液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物二叔丁基(3-溴-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(190mg)。
MS(ESI)M/Z:802.2[M+Na] +.
步骤C：在-10℃下，将氢氧化钾(1.7g)的水溶液(25mL)缓慢加入到乙醚(60mL)中，再加入1-甲基-1-亚硝基脲(3.0g，29.1mmol)反应液在-10℃下继续搅拌1小时。将反应液冷却至-78℃，水相冻住倒出有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，得到现制备的重氮甲烷的乙醚溶液(50mL)。
在室温下，将二叔丁基(3-溴-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(160mg，0.21mmol)和醋酸钯(41.4mg，0.18mmol)溶于四氢呋喃(4mL)，将反应体系置换氮气并冷却到-78℃，将现制备的重氮甲烷的乙醚溶液(50mL)缓慢加入到反应液中。反应体系缓慢回到室温，并在在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到化合物二叔丁基(3-溴-7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-基)氨基甲酸酯(85mg)。
MS(ESI)M/Z:816.3[M+Na] +.
步骤D：在室温条件下，将二叔丁基(3-溴-7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-基)氨基甲酸酯(75.0mg，0.09mmol)溶于混合溶液(氨水/异丙醇＝7/3，体积比，10mL)中，反应体系封管加热至120℃并搅拌24小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩得到粗产品化合物8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(45mg)。
MS(ESI)M/Z:575.2[M+H] +.
步骤E：在室温下，将8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(45mg，0.08mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速：20毫升/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品，减压冻干。得到(8-((3aS,4R,6R,6aR)-6-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺的两个异构体(实施例9-P1：7.1mg，实施例9-P2：5.5mg)。
实施例9-P1
HPLC：保留时间7.144分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:535.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.22(s,1H),7.91(s,1H),7.51(d,J＝8.3Hz,1H),7.27(s,1H),7.05(dd,J＝8.4,1.5Hz,1H),4.82-4.74(m,1H),4.58(dd,J＝8.3,5.7Hz,1H),4.17-3.95(m,1H),2.88(t,J＝6.4Hz,4H),2.53-2.27(m,3H),2.02-1.84(m,2H),1.79-1.65(m,1H),1.52-1.37(m,1H),1.20-0.99(m,2H).
实施例9-P2
HPLC：保留时间7.376分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:535.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.24(s,1H),7.98(s,1H),7.53(d,J＝8.3Hz,1H),7.28(s,1H),7.08(dd,J＝8.3,1.6Hz,1H),4.83-4.79(m,1H),4.51(dd,J＝8.2,6.0Hz,1H),4.07-3.96(m,1H),3.03-2.83(m,4H),2.59-2.46(m,2H),2.33(m,1H),2.13-2.02(m,1H),2.02- 1.83(m,1H),1.79-1.67(m,1H),1.40-1.29(m,1H),1.13-1.06(m,1H),1.06-0.95(m,1H).
实施例10
(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000087
反应路线:
Figure PCTCN2022106401-appb-000088
操作步骤：
步骤A：在室温条件下，将3-溴-2-氯-7-碘喹啉(150mg，0.43mmol)和环丙基甲胺(2mL)溶于乙醇(2mL)中，反应体系封管加热至120℃并搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到3-溴-N-(环丙基甲基)-7-碘喹啉-2-胺(70.0mg)。
MS(ESI)M/Z:402.8[M+H] +.
步骤B：在室温下，将8-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(20.0mg，0.059mmol)，3-溴-N-(环丙基甲基)-7-碘喹啉-2-胺(24.7mg，0.062mmol)和四乙基氯化铵(10.7mg， 0.065mmol)溶于N，N-二甲基甲酰胺(1.5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(2.6mg，0.012mmol)，N，N-二异丙基乙胺(38.1mg，0.29mmol)。反应液在70℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(20mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-6-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(25.0mg)。
MS(ESI)M/Z:308.2[2M+2H] 2+.
步骤C：在室温下，将8-((3aS,4R,6R,6aR)-6-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(25mg，0.04mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)环戊烷-1,2-二醇(6.0mg)。
MS(ESI)M/Z:575.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.98(s,1H),7.60(d,J＝8.4Hz,1H),7.52(s,1H),7.39(d,J＝8.3Hz,1H),6.69(t,J＝5.5Hz,1H),6.60(d,J＝5.5Hz,2H),6.46(s,2H),4.87(t,J＝6.1Hz,2H),4.84-4.74(m,1H),4.29(dd,J＝12.4,6.2Hz,1H),3.94(dd,J＝6.1Hz,1H),3.37-3.35(m,2H),2.95-2.79(m,4H),2.72(d,J＝6.4Hz,1H),2.44-2.36(m,2H),2.24-2.11(m,1H),1.91(dd,J＝23.1,10.7Hz,1H),1.30-1.14(m,1H),0.48-0.39(m,2H),0.35-0.22(m,2H).
实施例11
(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-氨基喹啉-7-基)氨基)甲基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000089
反应路线:
Figure PCTCN2022106401-appb-000090
操作步骤：
步骤A：在室温条件下，将8-((3aS,4R,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-氯-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶(405.0mg，0.67mmol)溶于氨水/异丙醇(8mL/3mL)中，反应体系封管加热至120℃并搅拌40小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS,4R,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(200.0mg)。
MS(ESI)M/Z:583.5[M+H] +.
步骤B：在室温下，将8-((3aS,4R,6R,6aR)-6-(((叔丁基二苯基硅基)氧基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-胺(170.0mg，0.29mmol)溶于四氢呋喃(5mL)中。将四丁基氟化铵(1.0M，0.58mL)缓慢滴加到反应体系中，反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到((3aR,4R,6R,6aS)-6-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)甲醇(73mg)。
MS(ESI)M/Z:345.3[M+H] +.
步骤C：在0℃条件下，将((3aR,4R,6R,6aS)-6-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)甲醇(88mg，0.26mmol)溶于吡啶(4mL)中，在氮气保护下，将三甲基氯硅烷(86.1mg，0.79mmol)缓慢 滴加到反应体系中，反应液在0℃下搅拌3小时。再将苯甲酰氯(43.8mg，0.31mmol)缓慢滴加到反应体系中，缓慢回到室温，反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入氨水(4mL)淬灭，反应液继续在室温下搅拌1小时后，混合液用二氯甲烷(50mL×2)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到N-(8-((3aS,4R,6R,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(70mg)。
MS(ESI)M/Z:449.3[M+H] +.
步骤D：在室温条件下，将N-(8-((3aS,4R,6R,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(50mg，0.11mmol)溶于二氯甲烷(3mL)中，在氮气保护下，将戴斯-马丁氧化剂(56.8mg，0.13mmol)缓慢滴加到反应体系中，反应液室温下搅拌2小时。
在TLC监测显示原料消失后，将反应过滤去除大部分固体后滤液浓缩得到粗产品N-(8-((3aS,4R,6S,6aR)-6-甲酰基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(70.0mg)直接用于下一步。
步骤E：在室温下，将N-(8-((3aS,4R,6S,6aR)-6-甲酰基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(49.9mg，0.11mmol)，2-氯喹啉-7-胺(29.9mg，0.17mmol)，乙酸(33.6mg，0.56mmol)和乙酸钾(32.9mg，0.34mmol)溶于甲醇(3mL)中。反应液在氮气保护和室温下搅拌2小时。将氰基硼氢化钠(21.1mg，0.33mmol)缓慢加入反应液中，反应液继续在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入乙酸乙酯(25mL)，再用饱和碳酸氢钠(20mL×2)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到N-(8-((3aS,4R,6R,6aR)-6-((2-氯喹啉-7-基)氨基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(45mg)。
MS(ESI)M/Z:609.4[M+H] +.
步骤F：在室温条件下，将N-(8-((3aS,4R,6R,6aR)-6-((2-氯喹啉-7-基)氨基)甲基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-5,6,7,8-四氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-4-基)苯甲酰胺(33.3mg，0.067mmol)溶于氨水/乙醇(20mL/4mL)中，反应体系在闷罐中密封加热至200℃并搅拌40小时。
LCMS监测显示60％原料消失后，将反应液减压蒸馏浓缩。所得残余物并未纯化，直接用于下一步。
MS(ESI)M/Z:486.2[M+H] +.
步骤G：在室温下，将N 7-(((3aR,4R,6R,6aS)-6-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)甲基)喹啉-2,7- 二胺(35mg，0.07mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-6,7-二氢环戊二烯并[4,5]吡咯并[2,3-d]嘧啶-8(5H)-基)-5-(((2-氨基喹啉-7-基)氨基)甲基)环戊烷-1,2-二醇(1.0mg)。
MS(ESI)M/Z:446.2[M+H] +.
1H NMR(400MHz,MeOD)δ7.98(s,1H),7.78(d,J＝8.9Hz,1H),7.35(d,J＝9.0Hz,1H),7.07(dd,J＝9.0,2.6Hz,1H),6.78(d,J＝2.6Hz,1H),6.74(d,J＝8.9Hz,1H),4.93-4.86(m,1H),4.60-4.46(m,1H),4.05(dd,J＝5.9,4.3Hz,1H),3.43-3.35(m,1H),3.28-3.21(m,1H),2.97-2.86(m,4H),2.55-2.45(m,2H),2.44-2.36(m,1H),2.36-2.26(m,1H),1.96-1.81(m,1H).
实施例12
(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((E)-2-(2-氨基喹啉-7-基)乙烯基)环戊烷-1,2-二醇三氟乙酸盐
Figure PCTCN2022106401-appb-000091
反应路线:
Figure PCTCN2022106401-appb-000092
操作步骤：
步骤A：在室温下，将4-氯-6,7-二氢-5H-吡咯并[2,3-D]嘧啶(775mg，5.0mmol)溶于 四氢呋喃(40mL)中。在室温下向反应体系中分批加入叔丁醇钾(559mg，5.0mmol)。反应液在室温下搅拌2小时。
原料消失后，反应液为白色浑浊液体。将反应液浓缩，旋干，加入N,N-二异丙基乙胺(20mL)，超声10分钟。将反应液过滤，滤饼即为4-氯-5,6-二氢吡咯并[2,3-d]嘧啶-7-化钾(湿重1.1g)。
1H NMR(400MHz,DMSO-d 6)δ7.38(s,1H),3.48(t,J＝8.6Hz,2H),2.66(t,J＝8.4Hz,2H).
步骤B：在室温下，将4-氯-5,6-二氢吡咯并[2,3-d]嘧啶-7-化钾(562mg，2.9mmol)和溶于N,N-二甲基甲酰胺(12mL)中。将(3aR,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲烷磺酸酯(910mg，2.9mmol)溶于N,N-二甲基甲酰胺(6mL)中。随后，在冰浴下向反应体系中缓慢加入上述溶液。反应液在25℃油浴下搅拌过夜。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(520mg)。
MS(ESI)M/Z:322.0[M+H] +.
步骤C：在室温下，将4-氯-7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(150mg，0.5mmol)，7-溴喹啉-2-胺(125mg，0.6mmol)和四乙基氯化铵(85.1mg，0.5mmol)溶于N,N-二甲基甲酰胺(6mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(21mg，0.1mmol)，N,N-二异丙基乙胺(0.4mL，2.42mmol)。反应液在100℃油浴下搅拌12小时。
TLC监测显示原料消失后，向反应液中加入水(20mL)淬灭，减压浓缩。混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(20mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)喹啉-2-胺(115mg)。
1H NMR(400MHz,DMSO-d 6)δ8.14(s,1H),7.84(d,J＝8.8Hz,1H),7.57(d,J＝8.2Hz,1H),7.37(s,1H),7.35-7.30(m,1H),6.70(d,J＝8.8Hz,1H),6.59(d,J＝15.8Hz,1H),6.54-6.40(m,3H),4.80(dd,J＝7.4,5.6Hz,1H),4.51-4.46(m,2H),3.86-3.68(m,2H),3.05(t,J＝9.2Hz,2H),2.78(dd,J＝16.2,7.8Hz,1H),2.08(d,J＝7.6Hz,2H),1.48(s,3H),1.25(s,3H).
步骤D在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)喹啉-2-胺(237mg，0.5mmol)，氨基甲酸叔丁酯(240mg，2.1mmol)溶于N,N-二甲基甲酰胺(5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入三(二亚苄基丙酮)二钯(47mg，0.05mmol)，2- 二环己基磷-2’,4’,6’-三异丙基联苯(25mg，0.05mmol)，碳酸铯(333mg，1.0mmol)。反应液在100℃微波下搅拌3小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(60mg)。
MS(ESI)M/Z:445.2[M+H] +.
步骤E：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(60mg，0.1mmol)溶于甲醇(2mL)中。随后，向上述溶液中加入4mol/L的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1mol/L的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((E)-2-(2-氨基喹啉-7-基)乙烯基)环戊烷-1,2-二醇三氟乙酸盐(12.6mg)。
MS(ESI)M/Z:405.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.30(d,J＝9.2Hz,1H),8.20(s,1H),7.85(d,J＝8.4Hz,1H),7.63(d,J＝8.4Hz,1H),7.54(s,1H),7.17(brs,2H),6.98(d,J＝9.2Hz,1H),6.64-6.55(m,2H),5.10-4.75(m,2H),4.45-4.35(m,1H),4.05-3.94(m,1H),3.86-3.70(m,3H),2.93-2.82(m,2H),2.71-2.65(m,1H),2.03-1.96(m,1H),1.63-1.54(m,1H).
实施例13
(1R,2S,3R,5S)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊烷-1,2-二醇及其异构体
Figure PCTCN2022106401-appb-000093
反应路线:
Figure PCTCN2022106401-appb-000094
操作步骤：
参考化合物1-6的方法制备化合物13-2。
步骤A：冰浴下，向4-氯吡咯并[2,3-d]嘧啶-7-酰胺钾(416mg，2.16mmol)的N,N-二甲基甲酰胺(2mL)溶液逐滴滴加(3aR,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基三氟甲烷磺酸酯(685mg，2.16mmol,制备方法参考化合物1-6)的N,N-二甲基甲酰胺(2mL)溶液。滴加完毕后，反应液在室温下搅拌过夜。
TLC检测显示新点生成，反应液倒入冰水中。反应液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相用饱和食盐水(10mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到4-氯-7-((3aS,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-7H-吡咯并[2,3-d]嘧啶(390mg)。
MS(ESI)M/Z:320.2[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.64(s,1H),7.30(d,J＝3.6Hz,1H),6.63(d,J＝3.6Hz,1H),6.03-5.89(m,1H),5.23-5.12(m,2H),5.06-4.97(m,2H),4.62(t,J＝6.7Hz,1H),2.83(dd,J＝12.6,6.4Hz,1H),2.53-2.30(m,2H),1.59(s,3H),1.32(s,3H).
步骤B：在室温下，将4-氯-7-((3aS,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-7H-吡咯并[2,3-d]嘧啶(330mg，1.03mmol)，7-溴喹啉-2-胺(250mg，1.13mmol)和四乙基氯化铵(188mg，1.13mmol)溶于N,N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(46mg，0.206mmol)，N,N-二异丙基乙胺(665mg，5.16mmol)。反应液在100℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR,4R,6aS)-6-(4-氯-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(130mg)。
MS(ESI)M/Z:462.2[M+H] +.
步骤C：在一个10mL的封管中，加入7-((E)-2-((3aR,4R,6aS)-6-(4-氯-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(130mg，0.28mmol)，浓氨水(4mL)和异丙醇(2mL)。反应液被密封，然后加热到110℃并且在此温度下搅拌过夜。
TLC监测显示原料消失，减压浓缩得到粗产物。粗产物用硅胶板层析纯化得到7-((E)-2-((3aR,4R,6aS)-6-(4-氨基-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(120mg)。
1H NMR(400MHz,MeOD)δ8.14(d,J＝9.2Hz,1H),8.11(s,1H),7.73(d,J＝8.3Hz,1H),7.58-7.48(m,2H),7.32(d,J＝3.6Hz,1H),6.91(d,J＝9.1Hz,1H),6.76-6.54(m,3H),5.13-5.01(m,2H),4.75(t,J＝6.9Hz,1H),3.09-2.94(m,1H),2.56-2.35(m,2H),1.57(s,3H),1.31(s,3H).
步骤D:在高压釜中，加入化合物7-((E)-2-((3aR,4R,6aS)-6-(4-氨基-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(35mg，0.079mmol)，10wt％钯碳(10mg)和乙醇(20mL)。反应体系被置换为氢气氛围，压力为15个大气压下加热到35℃，反应24小时后监测反应，发现有20％(摩尔分数)的产物和80％(摩尔分数)的原料。于是继续在此压力温度下搅拌两天。
LCMS监测显示原料已经全部消耗完。反应液被过滤，滤饼用乙醇洗3次。合并后的滤液减压浓缩得到粗产物。粗产物用硅胶板层析纯化得到7-(2-((3aR,4S,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙基)喹啉-2-胺(9mg)。
MS(ESI)M/Z:447.2[M+H] +.
1H NMR(400MHz,MeOD)δ8.08(s,1H),7.31(d,J＝3.5Hz,1H),7.20(d,J＝7.7Hz,1H),7.08(d,J＝7.7Hz,1H),6.97(s,1H),6.65(d,J＝3.5Hz,1H),5.34(t,J＝4.8Hz,1H),4.96(d,J＝2.9Hz,1H),4.51(t,J＝5.9Hz,1H),2.99-2.91(m,4H),2.72(t,J＝7.7Hz,2H),2.40-2.32(m,1H),2.06-2.00(m,2H),1.94-1.87(m,1H),1.84-1.76(m,1H),1.51(s,3H), 1.33(s,3H).
步骤E:在室温下，将化合物7-(2-((3aR,4S,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙基)喹啉-2-胺(9mg，0.02mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于1mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8-9。所得溶液经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速：20mL/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品，减压冻干。得到(1R,2S,5S)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊烷-1,2-二醇的两个异构体(实施例13-P1，1.3mg，收率15.8％；实施例13-P2，1mg)。
实施例13-P1:
HPLC:保留时间5.327分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:407.2[M+H] +.
1H NMR(400MHz,MeOD)δ8.06(s,1H),7.22(d,J＝3.6Hz,1H),7.04(d,J＝7.5Hz,1H),6.87(d,J＝7.6Hz,1H),6.83(s,1H),6.60(d,J＝3.6Hz,1H),4.89(d,J＝8.0Hz,1H),4.31(dd,J＝7.8,6.2Hz,1H),3.90-3.83(m,1H),2.84-2.77(m,2H),2.74-2.51(m,4H),2.45-2.34(m,1H),2.06-1.93(m,2H),1.78-1.67(m,1H),1.66-1.56(m,1H).
实施例13-P2:
HPLC:保留时间5.691分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:407.2[M+H] +.
1H NMR(400MHz,MeOD)δ8.06(s,1H),7.22(d,J＝3.6Hz,1H),7.08(d,J＝7.7Hz,1H),6.86(dd,J＝7.6,1.5Hz,1H),6.74(s,1H),6.60(d,J＝3.6Hz,1H),4.90-4.88(m,1H),4.32(dd,J＝7.8,6.2Hz,1H),3.91-3.83(m,1H),2.94-2.87(m,2H),2.75-2.58(m,2H),2.54(dd,J＝8.4,6.7Hz,2H),2.43-2.34(m,1H),2.06-1.92(m,2H),1.77-1.68(m,1H),1.65-1.57(m,1H).
实施例14
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000095
反应路线:
Figure PCTCN2022106401-appb-000096
操作步骤：
步骤A：在室温下，将4-氯-7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(630mg，1.96mmol)，氨基甲酸叔丁酯(1.15g，9.82mmol)溶于N,N-二甲基甲酰胺(10mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入三(二亚苄基丙酮)二钯(180mg，0.2mmol)，2-二环己基磷-2’,4’,6’-三异丙基联苯(93.6mg，0.2mmol)，碳酸铯(1.3g，3.93mmol)。反应液在100℃微波下搅拌3小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-6,7-二氢-5H-吡咯烷[2,3-d]嘧啶-4-胺(260mg)。
MS(ESI)M/Z:303.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.82(s,1H),6.00(s,2H),5.87(ddd,J＝17.4,10.4,7.2Hz,1H),5.15-4.96(m,2H),4.70(dd,J＝7.4,5.4Hz,1H),4.32-4.24(m,2H),3.59-3.44(m, 2H),2.73-2.67(m,2H),2.55-2.52(m,1H),1.98-1.84(m,2H),1.43(s,3H),1.21(s,3H).
步骤B：在室温下，将7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-6,7-二氢-5H-吡咯烷[2,3-d]嘧啶-4-胺(57mg，0.19mmol)，3-溴-7-碘喹啉-2-胺(79mg，0.23mmol)和四乙基氯化铵(34.6mg，0.21mmol)溶于N,N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(4.3mg，0.02mmol)，N,N-二异丙基乙胺(0.01mL，0.06mmol)。反应液在65℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)-3-溴喹啉-2-胺(20mg)。
MS(ESI)M/Z:523.2[M+H] +,525.2[M+2+H] +.
步骤C：在室温下，将化合物7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)乙烯基)-3-溴喹啉-2-胺(20mg，0.04mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(5.2mg)。
MS(ESI)M/Z:483.0[M+H] +,485.2[M+2+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.81(s,1H),7.59(d,J＝8.98Hz,1H),7.40(d,J＝6.4Hz,2H),6.61(s,2H),6.61-6.44(m,2H),5.94(s,2H),4.82(d,J＝5.2Hz,1H),4.68(d,J＝5.6Hz,1H),4.22(dd,J＝16.0,8.0Hz,1H),3.97(dd,J＝11.8,5.98Hz,1H),3.72(q,J＝5.6Hz,1H),3.52(t,J＝8.8Hz,2H),2.75-2.71(m,2H),2.61-2.59(m,1H),1.97-1.91(m,1H),1.56-1.48(m,1H).
实施例15
(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000097
反应路线:
Figure PCTCN2022106401-appb-000098
操作步骤：
步骤A:在一个三口瓶中，加入7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(50mg，0.164mmol)和四氢呋喃(1mL)。置换反应体系为氮气体系，然后向反应体系中逐滴滴加9-BBN(1.64mmol)的四氢呋喃溶液(0.5M，3.2mL)，滴加完毕后将反应液加热到60℃，反应1小时。反应也被冷却到室温，向反应液中滴加磷酸钾(174mg，0.82mmol)的水(2mL)溶液。反应混合物继续在室温下搅拌30分钟，然后向反应体系中加入3-溴-7-碘喹啉-2-胺(60mg，0.17mmol)和[1,1’-双(二苯基膦基)二茂铁]二氯化钯(10mg，0.014mmol)。再次置换反应体系为氮气体系，然后加热到60℃，搅拌过夜。TLC监测显示原料消失后，反应液被减压浓缩，所得混合物被用硅胶板层析纯化得到7-(2-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(55mg，收率63.9％)。
MS(ESI)M/Z:525.8[M+H] +.
步骤B:在室温下，将7-(2-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(55mg，0.104mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液被减压浓缩，然后用甲醇稀释同时加入7M的氨甲醇溶液调节pH值约等于8-9。混合液被加压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(18mg，收率36.0％)。
MS(ESI)M/Z:484.8[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.79(s,1H),7.57(d,J＝8.2Hz,1H),7.30(s,1H),7.10(d,J＝8.2Hz,1H),6.56(s,2H),5.92(s,2H),4.77(d,J＝5.6Hz,1H),4.40(d,J＝4.7Hz,1H),4.20-4.07(m,1H),3.93-3.82(m,1H),3.60-3.41(m,3H),2.79-2.68(m,4H),1.97-1.80(m,2H),1.80-1.63(m,1H),1.63-1.45(m,1H),1.31-1.11(m,1H).
实施例16
(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((1R)-2-(2-氨基喹啉 -7-基)环丙基)环戊烷-1,2-二醇及其异构体
Figure PCTCN2022106401-appb-000099
反应路线:
Figure PCTCN2022106401-appb-000100
操作步骤：
步骤A：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(300mg，0.65mmol)，二碳酸二叔丁酯(0.6mL，2.6mmol)和4-二甲氨基吡啶(16mg，0.13mmol)溶于四氢呋喃(10mL)中。随后，向上述溶液中加入三乙胺(0.36mL，2.6mmol)。反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。向粗产品中加入水(50mL)淬灭，混合液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(190mg)。
MS(ESI)M/Z:664.2[M+H] +.
步骤B：在-10℃下，将氢氧化钾(1.92g，30mL水)水溶液缓慢加入到乙醚(70mL)中，再加入1-甲基-1-亚硝基脲(3.5g，33.9mmol)反应液在-10℃下继续搅拌1小时。将反应液冷却至-78℃，水相冻住倒出有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，得到现制备的重氮甲烷的乙醚溶液(50mL)。
在室温下，将二叔丁基(7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-基)氨基甲酸酯(150mg，0.23mmol)和醋酸钯(50.8mg，0.23mmol)溶于四氢呋喃(4mL)，将反应体系置换氮气并冷却到-78℃，将现制备的重氮甲烷的乙醚溶液(70mL)缓慢加入到反应液中。反应体系缓慢回到室温，并在在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-氨基甲酸酯(85mg)。
步骤C：在室温下，将二叔丁基(7-((2R)-2-((3aR,4R,6R,6aS)-6-(4-氯-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-氨基甲酸酯(160mg，0.24mmol)，氨基甲酸叔丁酯(139mg，1.18mmol)溶于N,N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入三(二亚苄基丙酮)二钯(21.5mg，0.024mmol)，2-二环己基磷-2’,4’,6’-三异丙基联苯(11.2mg，0.024mmol)，碳酸铯(153mg，0.47mmol)。反应液在100℃微波下搅拌3小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((2R)-2-((3aR,4R,6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-氨基甲酸酯(62mg)。
MS(ESI)M/Z:659.2[M+H] +.
步骤D：在室温下，将化合物二叔丁基(7-((2R)-2-((3aR,4R,6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)环丙基)喹啉-2-氨基甲酸酯(62mg，0.094mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速:20毫升/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品得到(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((1R)-2-(2-氨基喹啉-7-基)环丙基)环戊烷-1,2-二醇的两个异构体(实施例16-P1：9.6mg，实施例16-P2：3.2mg)。
实施例16-P1
HPLC：保留时间5.495分钟。条件如下，柱子XBridge BEH C18 2.5μm，4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:419.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.81-7.74(m,2H),7.46(d,J＝8.2Hz,1H),7.11(s,1H),6.86(dd,J＝8.2,1.6Hz,1H),6.63(d,J＝8.8Hz,1H),6.32(s,2H),5.90(s,2H),4.83(s,1H),4.47(d,J＝4.7Hz,1H),4.13(dd,J＝18.0,8.0Hz,1H),3.96(t,J＝6.8Hz,1H),3.70(dd,J＝10.0,4.9Hz,1H),3.57-3.39(m,2H),2.70(dd,J＝17.7,6.3Hz,2H),1.96-1.80(m,2H),1.58-1.45(m,1H),1.36-1.22(m,1H),1.19-1.08(m,1H),1.02-0.91(m,2H).
实施例16-P2
HPLC:保留时间3.380分钟。条件如下，柱子ACQUITY UPLC BEH C18 1.7μm，2.1×150mm；柱温30℃；流动相水(0.05vol％TFA)和乙腈(0.05vol％TFA)；流速0.3毫升/分钟；梯度：5vol％乙腈(0.05vol％TFA)0.5分钟，9分钟内乙腈(0.05vol％TFA)升到95vol％，保持95vol％梯度3分钟。
MS(ESI)M/Z:419.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ7.79(t,J＝4.4Hz,2H),7.47(d,J＝8.2Hz,1H),7.12(s,1H),6.86(d,J＝8.2Hz,1H),6.64(d,J＝8.8Hz,1H),6.30(s,2H),5.92(s,2H),4.80(d,J＝5.6Hz,1H),4.47(d,J＝4.6Hz,1H),4.15(dd,J＝18.0,8.0Hz,1H),3.93(dd,J＝13.2,5.6Hz,1H),3.66(dd,J＝9.8,4.8Hz,1H),3.56-3.41(m,2H),2.76-2.70(m,2H),1.94-1.82(m,2H),1.59-1.45(m,1H),1.40-1.28(m,1H),1.17-1.07(m,1H),0.97-0.84(m,2H).
实施例17
(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000101
反应路线:
Figure PCTCN2022106401-appb-000102
操作步骤：
步骤A：在室温下，7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(50.0mg，0.17mmol)，3-溴-N-(环丙基甲基)-7-碘喹啉-2-胺(69.6mg，0.17mmol)和四乙基氯化铵(30.4mg，0.18mmol)溶于N,N- 二甲基甲酰胺(3mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(7.5mg，0.03mmol)，N,N-二异丙基乙胺(107.7mg，0.83mmol)。反应液在70℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(30mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)-3-溴-N-(环丙基甲基)喹啉-2-胺(30.0mg)。
MS(ESI)M/Z:577.0[M+H] +.
步骤B：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)-3-溴-N-(环丙基甲基)喹啉-2-胺(30mg，0.05mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5R)-3-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-5-((E)-2-(3-溴-2-((环丙基甲基)氨基)喹啉-7-基)乙烯基)环戊烷-1,2-二醇(6.0mg)。
MS(ESI)M/Z:537.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.31(s,1H),7.80(d,J＝8.4Hz,1H),7.58(d,J＝8.4Hz,1H),7.49(s,1H),7.35(dd,J＝8.4,1.5Hz,1H),6.68(t,J＝5.6Hz,1H),6.57-6.44(m,2H),5.94(s,2H),4.81(d,J＝5.3Hz,1H),4.68(d,J＝5.6Hz,1H),4.23(dd,J＝16.3,8.1Hz,1H),4.03-3.92(m,1H),3.78-3.68(m,1H),3.52(t,J＝9.0Hz,2H),3.37-3.33(m,2H),3.30-3.29(m,1H),2.76-2.69(m,2H),2.01-1.86(m,1H),1.53(dd,J＝22.9,9.9Hz,1H),1.24-1.21(m,1H),0.48-0.38(m,2H),0.33-0.25(m,2H).
实施例18
(1S,2R,3R,5R)-3-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇及其异构体
Figure PCTCN2022106401-appb-000103
反应路线:
Figure PCTCN2022106401-appb-000104
操作步骤：
步骤A：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)-3-溴喹啉-2-胺(160.0mg，0.31mmol)，二碳酸二叔丁酯(434.9mg，1.99mmol)和4-二甲氨基吡啶(7.6mg，0.06mmol)溶于四氢呋喃(8mL)中。随后，向上述溶液中加入三乙胺(250.9mg，2.5mmol)。反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。向粗产品中加入水(50mL)淬灭，混合液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((3aS,4R,6R,6aR)-6-((E)-2-(2-(二叔丁氧羰基)氨基)-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-基)亚氨基二碳酸酯(160mg)。
MS(ESI)M/Z:923.5[M+H] +.
步骤B：在-10℃下，将氢氧化钾(1.7g，25mL水)水溶液缓慢加入到乙醚(60mL)中，再加入1-甲基-1-亚硝基脲(3.0g，29.1mmol)反应液在-10℃下继续搅拌1小时。将反应液冷却至-78℃，水相冻住倒出有机相，有机相先用饱和食盐水(100mL)洗涤，然后用无水硫酸钠干燥，过滤，得到现制备的重氮甲烷的乙醚溶液(50mL)。
在室温下，将二叔丁基(7-((3aS,4R,6R,6aR)-6-((E)-2-(2-(二叔丁氧羰基)氨基)-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-基)亚氨基二碳酸酯(150mg，0.16mmol)和醋酸钯(32.8mg，0.15mmol)溶于四氢呋喃(4mL)，将反应体系置换氮气并冷却到-78℃，将现制备的重氮甲烷的乙醚溶液(50mL)缓慢加入到反应液中。反应体系缓慢回到室温，并在在室温下搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到二叔丁基(7-((3aS,4R,6R,6aR)-6-((1R)-2-(2-(二叔丁氧羰基)氨基)-3-溴喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶 -4-基)亚氨基二碳酸酯(90mg)。
MS(ESI)M/Z:937.6[M+H] +.
步骤C：在室温下，将二叔丁基(7-((3aS,4R,6R,6aR)-6-((1R)-2-(2-(二叔丁氧羰基)氨基)-3-溴喹啉-7-基)环丙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-基)亚氨基二碳酸酯(70mg，0.07mmol)溶于二氯甲烷(2mL)中。随后，向上述溶液中加入三氟乙酸(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速:20毫升/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品得到(1S,2R,3R,5R)-3-((1R)-2-(2-氨基-3-溴喹啉-7-基)环丙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇的两个异构体(实施例18-P1：10.2mg，实施例18-P2：4.9mg)。
实施例18-P1：
HPLC:保留时间6.532分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:497.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.22(s,1H),7.81(s,1H),7.50(d,J＝8.3Hz,1H),7.23(s,1H),7.02(dd,J＝8.4,1.7Hz,1H),4.45-4.15(m,1H),4.12(dd,J＝8.5,5.9Hz,1H),3.91(dd,J＝5.8,4.0Hz,1H),3.77-3.60(m,1H),3.59-3.49(m,1H),2.80(t,J＝8.9Hz,2H),2.28-2.02(m,1H),1.96-1.83(m,1H),1.73-1.56(m,1H),1.56-1.41(m,1H),1.24-1.15(m,1H),1.12-1.00(m,2H).
实施例18-P2:
HPLC:保留时间6.676分钟。条件如下，柱子XBridge BEH C18 2.5μm 4.6mm×150mm；柱温30℃；流动相水(10mM NH 4HCO 3)和乙腈；流速1毫升/分钟；梯度：5vol％乙腈0.5分钟，9分钟内乙腈升到95vol％，保持95vol％梯度2分钟。
MS(ESI)M/Z:497.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.24(s,1H),7.85(s,1H),7.53(d,J＝8.4Hz,1H),7.27(s,1H),7.07(dd,J＝8.4,1.6Hz,1H),4.48-4.18(m,1H),4.16-4.01(m,1H),3.89(dd,J＝5.8,4.0Hz,1H),3.83-3.66(m,1H),3.67-3.57(m,1H),2.99-2.65(m,2H),2.17-2.09(m,1H),2.09-2.00(m,1H),1.70-1.60(m,1H),1.57-1.47(m,1H),1.26-1.16(m,1H),1.11-1.03(m,1H),1.03-0.94(m,1H).
实施例19
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-3-甲基环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000105
反应路线:
Figure PCTCN2022106401-appb-000106
操作步骤：
步骤A:在一个三口瓶中，加入碘化亚铜(5.92g，31.08mmol)和无水四氢呋喃(100mL)。置换反应体系为氮气体系，然后冷却到0℃。向反应体系中逐滴滴加甲基锂(59.7mmol)的四氢呋喃溶液(1.6M，37.3mL)，滴加完毕后继续在此温度下搅拌10分钟，然后反应体系被降温到-78℃。向反应体系中滴加(3aR,6aR)-2,2-二甲基-3a,6a-二氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(4g，25.9mmol)的四氢呋喃溶液。滴加完毕后，反应体系继续在-78℃搅拌1小时。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(100mL×3次)萃取，合并有机相，有机相用饱和食盐水(50mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,6S,6aR)-2,2,6-三甲基二氢-4H-环戊二烯并[d][1,3]二氧杂环-4(5H)-酮(3.27g)。
1H NMR(400MHz,DMSO-d 6)δ4.47(d,J＝5.3Hz,1H),4.35(d,J＝5.3Hz,1H),2.66(dd,J＝18.3,8.3Hz,1H),2.38(dd,J＝11.1,4.1Hz,1H),1.98-1.87(m,1H),1.31(s,3H),1.27(s,3H),0.99(d,J＝7.5Hz,3H).
步骤B:在一个三口瓶中，加入(3aR,6S,6aR)-2,2,6-三甲基二氢-4H-环戊二烯并[d][1,3]二氧杂环-4(5H)-酮(500mg，2.94mmol)和三乙基氯硅烷(1mL，5.88mmol)的四氢呋喃(8mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至-78℃，然后逐滴滴加双(三甲基硅基)氨基锂的四氢呋喃溶液(1M，4.41mL，4.41mmol)。滴加完毕后，反应体系继续在0℃下搅拌20分钟。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相用饱和食盐水(30mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到三乙基(3aR,6S,6aR)-2,2,6-三甲基-6,6a-二氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)氧基)硅烷(660mg)。
1H NMR(400MHz,DMSO-d 6)δ4.77(dd,J＝6.1,1.5Hz,1H),4.67(d,J＝2.2Hz,1H),4.16(d,J＝6.1Hz,1H),2.53-2.52(m,1H),1.32(s,3H),1.24(s,3H),1.03-0.90(m,12H),0.67(q,6H).
步骤C:在一个单口瓶中，加入三乙基(3aR,6S,6aR)-2,2,6-三甲基-6,6a-二氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)氧基)硅烷(660mg，2.32mmol)，醋酸钯(155mg，0.696mmol)和二甲基亚砜(10mL)，然后体系置换为氧气氛围。反应液被加热到65℃，并在此温度下反应过夜。
TLC监测显示原料消失后，将反应液倒入冰水中。反应液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相依次用水(30mL×3次)和饱和食盐水(30mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,6aR)-2,2,6-三甲基-4H-环戊二烯并[d][1,3]二氧杂环-4(6aH)-酮(268mg)。
1H NMR(400MHz,DMSO-d 6)δ5.98(d,J＝1.4Hz,1H),5.07(d,J＝5.5Hz,1H),4.46(d,J＝5.5Hz,1H),2.13(s,3H),1.33(s,3H),1.28(s,3H).
步骤D:在一个三口瓶中，加入氯化锂(20mg，0.476mmol)，碘化亚铜(45mg，0.238mmol)和四氢呋喃(4mL)，然后体系置换为氮气氛围。反应液被冷却到0℃并在此温度下搅拌10分钟，然后向反应液中逐滴滴加三甲基氯硅烷(308.5mg，2.85mmol)和(3aR,6aR)-2,2,6-三甲基-4H-环戊二烯并[d][1,3]二氧杂环-4(6aH)-酮(400mg，2.38mmol)的四氢呋喃(2mL)溶液。滴加完毕后，反应液继续在0℃下搅拌20分钟。然后逐滴滴加乙烯基溴化镁(3.8mmol)的四氢呋喃溶液(1M，3.8mL)。所得反应液在0℃下继续搅拌30分钟。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相用饱和食盐水(30mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(3aR,6R,6aR)-2,2,6-三甲基-6-乙烯基二氢-4H-环戊二烯并[d][1,3]二氧杂环-4(5H)-酮(300mg)。
1H NMR(400MHz,DMSO-d 6)δ5.85(dd,J＝17.6,10.9Hz,1H),5.05(dd,J＝14.3,12.5Hz,2H),4.52(d,J＝5.0Hz,1H),4.24(d,J＝5.0Hz,1H),2.53-2.48(m,1H),2.30(d,J＝17.7Hz,1H),1.35(s,3H),1.30(s,3H),1.21(s,3H).
步骤E:在一个三口瓶中加入(3aR,6R,6aR)-2,2,6-三甲基-6-乙烯基二氢-4H-环戊二烯并 [d][1,3]二氧杂环-4(5H)-酮(700mg，3.57mmol)的四氢呋喃(10mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后逐滴滴加四氢铝锂(7.14mmol)的四氢呋喃溶液(1M，7.14mL)。滴加完毕后，反应体系继续在0℃下搅拌1小时。
TLC监测显示原料消失后，向反应液中缓慢加入十水合硫酸钠淬灭直至没有气泡冒出。然后加入四氢呋喃搅拌30分钟，过滤反应液。滤液被浓缩得到油状粗产物，用硅胶柱层析纯化得到(3aS,4S,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(400mg)。
1H NMR(400MHz,CDCl 3)δ5.70(dd,J＝17.6,11.0Hz,1H),5.08-4.95(m,2H),4.46(t,J＝5.7Hz,1H),4.33(dd,J＝5.4,0.8Hz,1H),4.09-3.94(m,1H),2.02-1.90(m,1H),1.59-1.52(m,1H),1.51(s,3H),1.36(s,3H),1.12(s,3H).
步骤F：在室温下，将4-氯-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(275mg，1.77mmol)和(3aS,4S,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(440mg，2.22mmol)溶于甲苯(5mL)中，反应体系被置换为氮气氛围。然后将2-(三丁基亚正膦基)乙腈(853mg，3.54mmol)的甲苯(2mL)溶液缓慢加入反应体系中。反应液加热到110℃，并在此温度下搅拌5小时。
TLC监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-((3aS,4R,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(160mg)。
MS(ESI)M/Z:336.0[M+H] +.
步骤G:在室温下，向高压反应釜中加入4-氯-7-((3aS,4R,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(130mg，0.39mmol)的乙醇(3mL)溶液。随后，在室温下向反应体系中缓慢加入氨水(30mL)。反应液在135℃油浴下搅拌2天。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS,4R,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(45mg)。
MS(ESI)M/Z:317.3[M+H] +.
步骤H：在室温下，将7-((3aS,4R,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(80mg，0.25mmol)，3-溴-7-碘喹啉-2-胺(88mg，0.25mmol)和四乙基氯化铵(45.6mg，0.28mmol)溶于N,N-二甲基甲酰胺(1mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(11.2mg，0.05mmol)，N,N-二异丙基乙胺(0.09mL，0.5mmol)。反应液在70℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水 硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2,4-三甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)-3-溴喹啉-2-胺(28mg)。
MS(ESI)M/Z:537.4[M+H] +.
步骤I：在室温下，将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2,4-三甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)-3-溴喹啉-2-胺(28mg，0.05mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化。纯化条件如下，制备柱welch XBC18，21.2×150mm，5μm(粒径)；流动相：水(含有10毫摩尔/升碳酸氢铵)和乙腈；流速：20毫升/分钟；梯度：在10分钟内，乙腈从40vol％升到60vol％；检测波长：214nm。收集产品得到(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-3-甲基环戊烷-1,2-二醇(4.7mg)。
MS(ESI)M/Z:497.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.81(s,1H),7.59(d,J＝9.0Hz,1H),7.41(d,J＝7.6Hz,2H),6.60(s,2H),6.53(d,J＝4.8Hz,2H),5.93(s,2H),4.85(d,J＝5.6Hz,1H),4.63(d,J＝5.6Hz,1H),4.34(t,J＝8.4Hz,1H),4.13(dd,J＝12.6,5.8Hz,1H),3.70(t,J＝5.6Hz,1H),3.52(t,J＝8.8Hz,2H),3.29-2.69(m,2H),1.81(dd,J＝13.2,9.4Hz,1H),1.70(dd,J＝13.2,9.4Hz,1H),1.17(s,3H).
实施例20
(1S,2R,3S,5R)-3-((E)-3-(2-氨基-3-溴喹啉-7-基)烯丙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000107
反应路线
Figure PCTCN2022106401-appb-000108
操作步骤：
步骤A：在一个三口瓶中，加入溴化亚铜二甲硫醚的络合物(411mg，2mmol)和四氢呋喃(80mL)。置换反应体系为氮气体系，然后冷却到-78℃。向反应体系中逐滴滴加乙烯基溴化镁(25mmol)的四氢呋喃溶液(1M，25mL)，滴加完毕后继续在此温度下搅拌20分钟，然后向反应体系中滴加(3aR,6aR)-2,2-二甲基-3a,6a-二氢-4H-环戊二烯并[d][1,3]二氧杂环-4-酮(3.08g，20mmol)，三甲基氯硅烷(4.34mL，40mmol)和六甲基膦酰三胺(9.2g，51.4mmol)的四氢呋喃溶液。滴加完毕后，反应体系继续在-78℃搅拌3小时，然后升到0℃继续搅拌1小时。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(500mL×3次)萃取，合并有机相，有机相用饱和食盐水(300mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到(6S)-6-烯丙基-2,2-二甲基二氢-4H-环戊二烯并[d][1,3]二氧杂环-4(5H)-酮(800mg)。
1H NMR(400MHz,CDCl 3)δ5.78-5.74(m,1H),5.17-5.09(m,2H),4.97(d,J＝5.4Hz,1H),4.37(d,J＝5.4Hz,1H),3.28-3.02(m,1H),2.82-2.48(m,1H),2.46-2.32(m,2H),2.25-2.00(m,1H),1.44(s,3H),1.39(s,3H).
步骤B：在一个三口瓶中加入(6S)-6-烯丙基-2,2-二甲基二氢-4H-环戊二烯并[d][1,3]二氧杂环-4(5H)-酮(1g，5.1mmol)的四氢呋喃(30mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后逐滴滴加四氢铝锂(10mmol)的四氢呋喃溶液(1M，10mL)。滴加完毕后，反应体系继续在0℃下搅拌1小时。
TLC监测显示原料消失后，向反应液中缓慢加入十水合硫酸钠淬灭。然后加入四氢呋喃搅拌30分钟，过滤反应液。滤液被浓缩得到油状粗产物，用硅胶柱层析纯化得到(4S,6S)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(300mg)。
1H NMR(400MHz,DMSO-d 6)δ5.81-5.68(m,1H),5.06-4.96(m,2H),4.37-4.30(m,2H),4.19(d,J＝5.7Hz,1H),3.93(dd,J＝10.8,4.6Hz,1H),2.04-1.93(m,1H),1.93-1.81(m,2H),1.80-1.70(m,1H),1.51-1.42(m,1H),1.37(s,3H),1.23(s,3H).
步骤C:在室温下，将4-氯-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(104mg，0.67mmol)和(4S,6S)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(200g，1.01mmol)溶于甲苯(2mL)中。将2-(三丁基亚正膦基)乙腈(482mg，2.0mmol)溶于甲苯(0.5mL)中。随后，在室温下向反应体系中缓慢加入2-(三丁基亚正膦基)乙腈溶液。反应液在110℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS,4R,6S,6aR)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-氯-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(164mg)。
MS(ESI)M/Z:336.0[M+H] +.
步骤D:在室温下，向高压反应釜中加入7-((3aS,4R,6S,6aR)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-氯-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(244mg，0.73mmol)的乙醇(3mL)溶液。随后，在室温下向反应体系中缓慢加入氨水(25mL)。反应液在135℃油浴下搅拌2天。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS,4R,6S,6aR)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(90mg)。
MS(ESI)M/Z:317.2[M+H] +.
步骤E:在室温下，将7-((3aS,4R,6S,6aR)-6-烯丙基-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(70mg，0.22mmol)，3-溴-7-碘喹啉-2-胺(77mg，0.22mmol)和四乙基氯化铵(40mg，0.24mmol)溶于N,N-二甲基甲酰胺(1mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(5mg，0.02mmol)，N,N-二异丙基乙胺(0.08mL，0.484mmol)。反应液在65℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-3-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)丙-1-烯-1-基)-3-溴喹啉-2-胺(54mg)。
MS(ESI)M/Z:537.0[M+H] +.
步骤F：在室温下，将7-((E)-3-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)丙-1-烯-1-基)-3-溴喹啉-2-胺(62mg，0.12mmol)溶于甲醇(0.5mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8-9。混合液用二氯甲烷(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S,2R,3S,5R)-3-((E)-3-(2-氨基-3-溴喹啉-7-基)烯丙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(14.5mg)。
MS(ESI)M/Z:497.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.31(s,1H),7.78(s,1H),7.58(d,J＝8.8Hz,1H),7.41-7.35(m,2H),6.63-6.56(m,3H),6.47-6.42(m,1H),5.91(s,2H),4.81(d,J＝5.6Hz,1H),4.47(d,J＝4.8Hz,1H),4.16(dd,J＝17.8,7.8Hz,1H),3.91(dd,J＝13.6,5.8Hz,1H),3.59(dd,J＝9.9,4.8Hz,1H),3.54-3.41(m,2H),2.73-2.69(m,1H),2.44-2.38(m,2H),2.25-2.15(m,1H),1.97-1.83(m,2H),1.28-1.19(m,1H).
实施例21
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000109
反应路线:
Figure PCTCN2022106401-appb-000110
操作步骤：
步骤A：在室温下，将4-氯-7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(210mg，0.65mmol)，1,1’-二(二苯膦基)二茂铁二氯化钯(II)(95.1mg，0.13mmol)和碳酸钾(179.6mg，1.3mmol)溶于二氧六环/水(5mL/0.5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入2,4,6-三甲基-1,3,5,2,4,6-三氧杂硼环己烷(246.9mg，1.96mmol)，反应体系封管加热至110℃并搅拌18小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-甲基-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(160.0mg)。
MS(ESI)M/Z:302[M+H] +.
步骤B：在室温下，将7-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-4-甲基-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(40mg，0.13mmol)和3-溴-7-碘喹啉-2-胺(42.1mg，0.12mmol)溶于N,N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(5.8mg，0.026mmol)，N,N-二异丙基乙胺(83.8mg，0.65mmol)。反应液在70℃油浴下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(50mL)淬灭，混合液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到3-溴-7-((E)-2-((3aR,4R,6R,6aS)-2,2-二甲基-6-(4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(23mg)。
MS(ESI)M/Z:522[M+H] +.
步骤C：在室温下，将3-溴-7-((E)-2-((3aR,4R,6R,6aS)-2,2-二甲基-6-(4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙烯基)喹啉-2-胺(36.0mg，0.07mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-)4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(11.7mg)。
MS(ESI)M/Z:482.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.25(s,1H),8.13(s,1H),7.56(d,J＝8.4Hz,1H),7.47(s,1H),7.42(dd,J＝8.4,1.6Hz,1H),6.62(d,J＝15.9Hz,1H),6.49(dd,J＝15.8,7.8Hz,1H),4.56-4.31(m,1H),4.29-4.14(m,1H),3.91(t,J＝6.0Hz,1H),3.77(t,J＝8.7Hz,2H),3.04(t,J＝8.6Hz,2H),2.88-2.71(m,1H),2.21(s,3H),2.19-2.07(m,1H),1.71(dt,J＝12.9,10.3Hz,1H).
实施例22
(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-3-甲基环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000111
反应路线:
Figure PCTCN2022106401-appb-000112
操作步骤：
步骤A：在一个三口瓶中加入7-((3aS,4R,6R,6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-6,7-二氢-5H-吡咯并[2,3-d]嘧啶-4-胺(45mg，0.14mmol)的四氢呋喃(0.5mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(1.4mmol)的四氢呋喃溶液(0.5M，2.8mL)，滴加完毕后，反应液被加热到60℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(297mg，1.4mmol)的水溶液(0.5mL)。反应液在室温下搅拌30分钟后，加入3-溴-7-碘喹啉-2-胺(98mg，0.28mmol)和PdCl 2(dppf)(10mg，0.014mmol)的四氢呋喃(1.2mL)溶液。最后所得的反应混合液被加热到60℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到7-(2-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2,4-三甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙基)-3-溴喹啉-2-胺(45mg)。
MS(ESI)M/Z:539.0[M+H] +.
步骤B：在室温下，将7-(2-((3aR,4S,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2,4-三甲基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)乙基)-3-溴喹啉-2-胺(45mg，0.083mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于3mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8-9。所得溶液经制备型高效液相色谱纯化得到(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6H-二氢-7H-吡咯并[2,3-d]嘧啶-7(6H)-基)-3-甲基环戊烷-1,2-二醇(8.4mg)。
MS(ESI)M/Z:499.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.79(s,1H),7.57(d,J＝8.3Hz,1H),7.30(s,1H),7.10(dd,J＝8.2,1.6Hz,1H),6.55(s,2H),5.91(s,2H),4.81(d,J＝5.7Hz,1H),4.37(d,J＝5.1Hz,1H),4.25(dd,J＝7.8Hz,1H),4.08(dd,J＝7.5Hz,1H),3.58-3.53(m,1H),3.53-3.42(m,2H),2.73-2.69(m,2H),2.65-2.60(m,2H),1.69-1.56(m,2H),1.55-1.48(m,2H),1.04(s,3H).
实施例23
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000113
制备方法参考实施例21，最后得到目标产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-甲基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇。
MS(ESI)M/Z:438.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.14(d,J＝2.2Hz,2H),7.60(d,J＝9.0Hz,1H),7.44-7.38(m,2H),6.70(s,2H),6.50-6.45(m,2H),4.82-4.71(m,2H),4.35(dd,J＝17.2,8.0Hz,1H),4.02(dd,J＝12.4,5.8Hz,1H),3.74(q,J＝5.6Hz,1H),3.65(dd,J＝13.8,8.8Hz,2H),2.96(t,J＝8.5Hz,2H),2.67-2.60(m,1H),2.13(s,3H),1.98(dt,J＝12.8,8.0Hz,1H),1.62-1.51(m,1H).
实施例24
(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000114
反应路线:
Figure PCTCN2022106401-appb-000115
操作步骤：
步骤A:在室温下，将4-氯-9H-嘧啶并[4,5-b]吲哚(450mg，2.2mmol)和(3aS,4S,6S,6aR)-2,2-二甲基-6-烯丙基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(806mg，4.4mmol)的甲苯(5mL)混合物加入到20mL的封管中。反应混合物被置换氮气体系，随后，在室温下向反应体系中缓慢滴加2-(三丁基亚正膦基)乙腈(2.12g，8.8mmol)的甲苯(3mL)溶剂。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，反应液被水淬灭。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-9H-嘧啶并[4,5-b]吲哚(360mg)。
MS(ESI)M/Z:370.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.87(s,1H),8.38(d,J＝7.8Hz,1H),7.97(d,J＝8.4Hz,1H),7.72(dd,J＝11.5,4.1Hz,1H),7.52(t,J＝7.6Hz,1H),6.02(td,J＝10.3,5.2Hz,1H),5.42(d,J＝4.9Hz,1H),5.30(dd,J＝7.3,5.1Hz,1H),5.15(dd,J＝31.7,13.8Hz,2H),4.76(t,J＝7.0Hz,1H),2.90-2.81(m,1H),2.74(d,J＝12.1Hz,1H),2.29(d,J＝6.1Hz,1H),1.53(s,3H),1.22(s,3H).
步骤B:在室温下，向20mL的封管反应器中加入4-氯-9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-9H-嘧啶并[4,5-b]吲哚(360mg，0.98mmol)的异丙醇(3mL)溶液。随后，在室温下向反应体系中缓慢加入氨水(6mL)。密封封管，反应液在120℃油浴下搅拌过夜。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残余物用硅胶柱层析纯化得到9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(300mg)。
MS(ESI)M/Z:351.2[M+H] +.
步骤C：在一个三口瓶中加入9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(50mg，0.14mmol)的四氢呋喃(0.5mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(1.4mmol)的四氢呋喃溶液(0.5M，2.8mL)，滴加完毕后，反应液被加热到60℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(148mg，0.7mmol)的水溶液(0.5mL)。反应液在室温下搅拌30分钟后，加入3-溴-7-碘喹啉-2-胺(50mg，0.14mmol)和PdCl 2(dppf)(10mg，0.014mmol)的四氢呋喃(1.2mL)溶液。最后所得的反应混合液被加热到60℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到9-((3aS,4R,6S,6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧醇-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(70mg，纯度60％)。
MS(ESI)M/Z:573.0[M+H] +，575.0[M+H+2] +.
步骤D：在室温下，将9-((3aS,4R,6S,6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧醇-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(70mg，0.12mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物溶于3mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8-9。所得溶液经制备型高效液相色谱纯化得到最终产物(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)环戊烷-1,2-二醇(9.6mg)。
MS(ESI)M/Z:533.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.36-8.31(m,2H),8.28(s,1H),7.60(t,J＝8.1Hz,2H),7.40(t,J＝7.7Hz,1H),7.35(s,1H),7.25(t,J＝7.5Hz,1H),7.21-7.11(m,3H),6.57(s,2H),5.11-5.00(m,1H),4.73(dd,J＝10.9,4.5Hz,2H),4.63(d,J＝5.4Hz,1H),3.99-3.87(m,1H),2.85-2.71(m,2H),2.14-1.95(m,4H),1.86-1.73(m,1H).
实施例25
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000116
反应路线:
Figure PCTCN2022106401-appb-000117
操作步骤：
步骤A：在室温下，将9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(50mg，0.14mmol)，3-溴-7-碘喹啉-2-胺(60.0mg，0.17mmol)和四乙基氯化铵(26.0mg，0.16mmol)溶于N,N-二甲基甲酰胺(3mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(6.4mg，0.03mmol)，N,N-二异丙基乙胺(0.124mL，0.75mmol)。反应液在70℃下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物9-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(35mg)。
MS(ESI)M/Z:571.0[M+H] +.
步骤B：在室温下，将9-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(35mg，0.06mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)环戊烷-1,2-二醇(15.0mg)。
MS(ESI)M/Z:531.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.39-8.29(m,3H),7.67(d,J＝8.3Hz,1H),7.62(d,J＝8.5Hz,1H),7.51-7.39(m,3H),7.27(t,J＝7.6Hz,1H),7.21(s,2H),6.74-6.59(m,4H),5.24-5.07(m,1H),4.93-4.86(m,2H),4.78-4.64(m,1H),4.25-4.07(m,1H),2.90-2.76(m,1H),2.37-2.28(m,1H),2.23-2.13(m,1H).
实施例26
(1R,2S,3R,5S)-3-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊 烷-1,2-二醇
Figure PCTCN2022106401-appb-000118
反应路线:
Figure PCTCN2022106401-appb-000119
操作步骤：
步骤A：在室温下，将9-((3aS,4R,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(100mg，0.28mmol)，7-碘喹啉-2-胺(91.8mg，0.34mmol)和四乙基氯化铵(51.8mg，0.31mmol)溶于N,N-二甲基甲酰胺(3mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(12.8mg，0.057mmol)，N,N-二异丙基乙胺(0.25mL，1.512mmol)。反应液在70℃下搅拌18小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物9-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(130mg)。
MS(ESI)M/Z:493.2[M+H] +.
步骤B：在室温下，将钯碳(236.0mg，2.2mmol)溶于乙醇(5mL)中。反应体系抽真空 换氢气多次。在氢气氛围下加入9-((3aS,4R,6R,6aR)-6-((E)-2-(2-氨基喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(110mg，0.22mmol)。反应液在室温下搅拌18小时。
LCMS监测显示原料消失后，过滤浓缩，混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到化合物9-((3aS,4R,6S,6aR)-6-(2-(2-氨基喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(90mg)。
MS(ESI)M/Z:495.2[M+H] +.
步骤C：在室温下，将9-((3aS,4R,6S,6aR)-6-(2-(2-氨基喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊二烯并[d][1,3]二氧杂-4-基)-9H-嘧啶并[4,5-b]吲哚-4-胺(70mg，0.14mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。往残余物里加入甲醇(1mL)再向反应液中加入氨水溶液调节pH值约等于8-9。混合液经制备型高效液相色谱纯化得到最终产物(1R,2S,3R,5S)-3-(4-氨基-9H-嘧啶并[4,5-b]吲哚-9-基)-5-(2-(2-氨基喹啉-7-基)乙基)环戊烷-1,2-二醇(48.5mg)。
MS(ESI)M/Z:555.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.33(d,J＝7.7Hz,1H),8.29(s,1H),7.82(d,J＝8.8Hz,1H),7.62(d,J＝8.3Hz,1H),7.53(d,J＝8.1Hz,1H),7.44-7.37(m,1H),7.28(d,J＝6.6Hz,1H),7.25(d,J＝7.3Hz,1H),7.18(s,2H),7.06(dd,J＝8.1,1.5Hz,1H),6.68(d,J＝8.8Hz,1H),6.33(s,2H),5.13-5.01(m,1H),4.81-4.69(m,2H),4.67-4.57(m,1H),3.99-3.88(m,1H),2.87-2.64(m,2H),2.19-1.91(m,4H),1.89-1.72(m,1H).
实施例27
(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000120
反应路线:
Figure PCTCN2022106401-appb-000121
操作步骤：
步骤A:室温下，在一个单口瓶中，加入乙腈(50mL)，4-氨基-6-氯嘧啶-5-醇(437mg，3mmol)和碳酸铯(1955mg，6mmol)，1，2-二溴乙烷(1127mg，6mmol)，然后加热到65℃搅拌16小时。
反应液过滤，滤液减压浓缩。所得粗品用硅胶柱层析纯化得到4-氯-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪(235mg)。
MS(ESI)M/Z:172.0[M+H] +.
步骤B:在封管中，将4-氯-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪(205mg，1.2mmol)和(3aS，4S，6R，6aR)-2,2,6-三甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英-4-醇(442mg，2.4mmol)溶于甲苯(3mL)。将氰基亚甲基三正丁基膦(869mg，3.6mmol)溶于甲苯(1mL)中，在氮气氛围下滴加上述液体，反应液在110℃下搅拌18小时。
反应液减压浓缩，所得粗品用硅胶柱层析纯化得到4-氯-8-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪(295mg)。
MS(ESI)M/Z:338.2[M+H] +.
步骤C:在室温下，将4-氯-8-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪(295mg，0.872mmol)，氨基甲酸叔丁酯(202.5mg，1.74mmol)溶于无水甲苯(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入Brettphos Pd G2(73.6mg，0.087mmol)，碳酸铯(566.8mg，1.74mmol)。反应液在110℃下搅拌18小时。
反应液减压浓缩，所得粗品用硅胶柱层析纯化得到8-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(105mg)。
MS(ESI)M/Z:319.2[M+H] +.
步骤D:在三口瓶中，在氮气保护下加入无水四氢呋喃(5mL)，8-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(115mg，0.35mmol)，然后滴加9-BBN(299mg，2.45mmol)。反应液加热到55℃，搅拌1小时。反应液再冷却室温，加入磷酸钾(520mg，2.45mmol，溶于1.5mL水中)，搅拌20分钟。加入3-溴-7-碘喹啉-2-胺(147mg，0.42mmol，溶于2mL无水四氢呋喃中)，PdCl 2(dppf)(55mg，0.07mmol)。反应加热到55℃，搅拌16小时。反应冷却至室温，过滤，滤液浓缩干燥。所得粗品用硅胶柱层析纯化得到8-((3aS，4R，6S，6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(105mg)。
MS(ESI)M/Z:541.1[M+H] +.
步骤E:室温下，在一个单口瓶中，加入甲醇(2mL)，8-((3aS，4R，6S，6aR)-6-(2-(2-氨基-3-溴喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(81mg，0.15mmol)，滴加4N盐酸甲醇溶液(2mL)，然后搅拌2小时。LCMS监测显示原料消失后，反应液减压浓缩。所得粗品用反相色谱纯化得到(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)环戊烷-1,2-二醇(32mg)。
MS(ESI)M/Z:501.0[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.12(s,1H),7.82(s,1H),7.54–7.42(m,2H),7.17(d,J＝8.3Hz,1H),5.34(s,2H),4.69(s,2H),4.45–4.33(m,1H),4.28–4.18(m,2H),3.92(t,J＝7.3Hz,1H),3.81–3.70(m,1H),3.58–3.42(m,2H),2.92-2.82(m,2H),2.15-1.93(m,3H),1.77-1.70(m,1H),1.40-1.36(m,1H).
实施例28
(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000122
反应路线:
Figure PCTCN2022106401-appb-000123
操作步骤：
步骤A:在单口瓶中，加入无水DMF(2mL)，8-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(63mg，0.2mmol)，3-溴-7-碘喹啉-2-胺(84mg，0.24mmol)，醋酸钯(10mg，0.04mmol)，TEAC(66mg，0.4mmol)，DIPEA(51mg，0.394mmol)。体系转换成氮气，加热到70℃，搅拌4小时。反应冷却至室温，过滤，滤液浓缩干燥。所得粗品用硅胶柱层析纯化得到8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(25mg)。
MS(ESI)M/Z:539.1[M+H] +.
步骤B:室温下，在一个单口瓶中，加入甲醇(2mL)，8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-7,8-二氢-6H-嘧啶[5,4-b][1,4]恶嗪-4-胺(25mg，0.05mmol)，滴加4N盐酸甲醇溶液(2mL)，然后搅拌2小时。LCMS监测显示原料消失后，反应液减压浓缩。粗产物被溶于3mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8～9。所得溶液用反相色谱纯化得到(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)环戊烷-1,2-二醇(6.5mg)。
MS(ESI)M/Z:499.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.25(s,1H),7.69(s,1H),7.56(d,J＝8.4Hz,1H),7.47(s,1H),7.42(dd,J＝8.4,1.5Hz,1H),6.63(d,J＝15.9Hz,1H),6.49(dd,J＝15.8,7.7Hz,1H),4.92-4.84(m,1H),4.20-4.19(m,2H),4.15(dd,J＝16.4,9.8Hz,1H),3.91(t,J＝6.4Hz,1H),3.63–3.52(m,2H),2.79-2.75(m,1H),2.10-2.02(m,1H),1.74-1.58(m,1H).
实施例29
(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)-3-甲基环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000124
制备方法参考实施例27，最后得到目标产物(1S,2R,3S,5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢-8H-嘧啶[5,4-b][1,4]恶嗪-8-基)-3-甲基环戊烷-1,2-二醇。
MS(ESI)M/Z:515.3[M+H] +.
1H NMR(400MHz,MeOD)δ8.26(s,1H),7.67(s,1H),7.55(d,J＝8.2Hz,1H),7.38(s,1H),7.17(dd,J＝8.3,1.6Hz,1H),4.99-4.88(m,1H),4.26-4.22(m,1H),4.22-4.13(m,2H),3.75(d,J＝6.3Hz,1H),3.56–3.43(m,2H),2.85–2.72(m,2H),1.86–1.66(m,3H),1.66–1.56(m,1H),1.18(s,3H).
实施例30
(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000125
反应路线:
Figure PCTCN2022106401-appb-000126
操作步骤：
步骤A:在室温下，将8-((3aS，4R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(53mg，0.156mmol)，7-溴-3-氯喹啉-2-胺(40mg，0.156mmol，参考中间体Int-1的合成方法制备)和四乙基氯化铵(52.2mg，0.315mmol)溶于N，N-二甲基甲酰胺(1mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(7mg，0.03mmol)，N，N-二异丙基乙胺(40mg，0.312mmol)。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(63mg)。
MS(ESI)M/Z:517.2[M+H] +.
步骤B：在室温下，将8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(63mg，0.122mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩，纯化得到最终产物(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇(33.9mg)。
MS(ESI)M/Z:477.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.15(s,1H),7.97(s,1H),7.61(d,J＝8.8Hz,1H),7.43(d,J＝7.6Hz,2H),6.69(s,2H),6.59(s,2H),6.45(s,2H),4.92–4.83(m,2H),4.81–4.75(m,1H),4.29(dd,J＝12.4,6.4Hz,1H),3.93(dd,J＝11.8,5.8Hz,1H),2.95–2.78(m,4H),2.75-2.70(m,1H),2.42-2.39(m,2H),2.23–2.15(m,1H),1.93–1.88(m,1H).
实施例31
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-氟喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000127
反应路线:
Figure PCTCN2022106401-appb-000128
操作步骤：
步骤A:在室温下，将7-溴-3-氟喹啉(900mg，4mmol)溶于二氯甲烷(25mL)中。在0℃下缓慢加入间氯过苯甲酸(2.75g，16mmol)，反应液在室温下搅拌12小时。
LCMS监测显示原料消失后，0℃下向反应液中加入水(50mL)淬灭，然后缓慢加入氯化铵的饱和水溶液调节pH至7，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-溴-3-氟喹啉1-氧化物(850mg)。
MS(ESI)M/Z:242.2[M+H] +.
步骤B:在室温下，将7-溴-3-氟喹啉1-氧化物(850mg，3.54mmol)，溶于三氯甲烷(15mL)中。然后在0℃下缓慢加入三氯氧磷(15mL)，反应体系抽真空换氮气多次。反应液在70℃油浴下搅拌3小时。
LCMS监测显示原料消失后，减压浓缩。加入二氯甲烷(30mL)，然后用饱和碳酸氢钠溶液调节pH至7，混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(30mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-溴-2-氯-3-氟喹啉(700mg)。
MS(ESI)M/Z:259.9[M+H] +.
步骤C:在室温下，将7-溴-2-氯-3-氟喹啉(745mg，2.86mmol)溶于乙醇(10mL)和氨水(20mL)中。随后反应液置于焖罐中，在100℃下搅拌20小时。
LCMS监测显示原料消失后，减压浓缩，混合液用二氯甲烷(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓 缩。所得残余物用硅胶柱层析纯化得到7-溴-3-氯喹啉-2-胺(530mg)。
MS(ESI)M/Z:241.0[M+H] +.
步骤D:在室温下，将8-((3aS，4R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(125mg，0.36mmol)，7-溴-2-氯-3-氟喹啉(86mg，0.36mmol)和四乙基氯化铵(65mg，0.39mmol)溶于N，N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(16mg，0.07mmol)，N,N-二异丙基乙胺(0.13mL，0.786mmol)。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，减压浓缩。混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-氟喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(87mg)。
MS(ESI)M/Z:501.2[M+H] +.
步骤E：在室温下，将8-((3aS，4R，6R，6aR)-6-((E)-2-(2-氨基-3-氟喹啉-7-基)乙烯基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(87mg，0.174mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于2mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8～9。所得溶液经制备型高效液相色谱纯化得到最终产物(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-氟喹啉-7-基)乙烯基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-基)环戊烷-1,2-二醇(23.7mg)。
MS(ESI)M/Z:461.2[M+H] +.
1H NMR(400MHz,MeOD)δ7.98(s,1H),7.77(d,J＝11.8Hz,1H),7.60(d,J＝8.9Hz,1H)7.44–7.40(m,2H),6.74(s,2H),6.59-6.53(m,2H),6.46(s,2H),4.86(dd,J＝5.8,2.9Hz,2H),4.83–4.75(m,1H),4.29(dd,J＝12.5,6.2Hz,1H),3.93(dd,J＝6.0Hz,1H),2.96-2.80(m,4H),2.78-2.65(m,1H),2.45–2.35(m,2H),2.26-2.14(m,1H),1.97-1.85(m,1H).
实施例32
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-氟喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000129
制备方法参考实施例31，最后得到目标产物(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-氟喹啉 -7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇。
MS(ESI)M/Z:423.2[M+H] +.
1H NMR(400MHz,MeOD)δ7.85(s,1H),7.65(d,J＝11.3Hz,1H),7.57(d,J＝8.4Hz,1H),7.49(s,1H),7.41(d,J＝8.3Hz,1H),6.63-6.57(m,1H),6.48-6.38(m,1H),4.33-4.25(m,1H),4.15–4.11(m,1H),3.90–3.86(m,1H),3.72–3.61(m,2H),2.89-2.81(m,2H),2.81-2.74(m,1H),2.16–2.08(m,1H),1.70-1.58(m,1H).
实施例33
(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000130
反应路线:
Figure PCTCN2022106401-appb-000131
操作步骤：
步骤A:在室温下，将7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(60mg，0.2mmol)，7-溴-3-氯喹啉-2-胺(51mg，0.2mmol)和四乙基氯化铵(36.3mg，0.22mmol)溶于N，N-二甲基甲酰胺(1.5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(9mg，0.04 mmol)，N，N-二异丙基乙胺(51.6mg，0.4mmol)。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR，4R，6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)-3-氯喹啉-2-胺(40mg)。
MS(ESI)M/Z:479.0[M+H] +.
步骤B：在室温下，将7-((E)-2-((3aR，4R，6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)-3-氯喹啉-2-胺(40mg，0.084mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S，2R，3R，5R)-3-((E)-2-(2-氨基-3-氯喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(10.8mg)。
MS(ESI)M/Z:220.6[1/2M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.14(s,1H),7.81(s,1H),7.60(d,J＝8.6Hz,1H),7.40(s,2H),6.69(s,2H),6.58–6.42(m,2H),5.94(s,2H),4.83(d,J＝5.2Hz,1H),4.69(d,J＝5.4Hz,1H),4.26–4.15(m,1H),4.05-3.95(m,1H),3.78-3.68(m,1H),3.52(t,J＝8.6Hz,2H),2.79–2.68(m,2H),2.65-2.58(m,1H),1.98–1.90(m,1H),1.57–14.9(m,1H).
实施例34
(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5-甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000132
反应路线:
Figure PCTCN2022106401-appb-000133
操作步骤：
步骤A:在一个三口瓶中，加入2-(4,6-二氯嘧啶-5-基)乙酸甲酯(2g，9.1mmol)的四氢呋喃(100mL)溶液。置换反应体系为氮气体系，然后冷却到-78℃。向反应体系中逐滴滴加双三甲基硅基胺基锂(18.2mmol)的四氢呋喃溶液(1M，18.2mL)，滴加完毕后继续在此温度下搅拌1小时，然后向反应体系中滴加碘甲烷(2.58g，18.2mmol)，滴加完毕后，反应体系升到0℃继续搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(200mL×3次)萃取，合并有机相，有机相用饱和食盐水(200mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到2-(4,6-二氯嘧啶-5-基)丙酸甲酯(1.54g)。
MS(ESI)M/Z:234.9[M+H] +.
步骤B:在一个三口瓶中，加入2-(4,6-二氯嘧啶-5-基)丙酸甲酯(1g，4.27mmol)的四氢呋喃(30mL)溶液。置换反应体系为氮气体系，然后冷却到零下78℃。向反应体系中逐滴滴加二异丁基氢化铝(12.8mmol)的四氢呋喃溶液(1.5M，8.5mL)，滴加完毕后，反应体系缓慢升到室温继续搅拌过夜。
LCMS监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相用饱和食盐水(50mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化得到2-(4,6-二氯嘧啶-5-基)丙烷-1-醇(450mg)。
MS(ESI)M/Z:207.2[M+H] +.
步骤C:在三口瓶中，将2-(4,6-二氯嘧啶-5-基)丙烷-1-醇(540mg，2.62mmol)和4-二甲氨基吡啶(32mg，0.262mmol)溶于二氯甲烷(420mL)中。在0℃下加入甲磺酰氯(750mg，6.55mmol)和三乙胺(1.19g，11.8mmol)，反应液在0℃到室温下搅拌3小时。
LCMS监测显示原料消失后，减压浓缩。混合液用二氯甲烷(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到2-(4,6-二氯嘧啶-5-基)甲基磺酸丙酯(700mg)。
MS(ESI)M/Z:285.0[M+H] +.
步骤D:在室温下，将2-(4,6-二氯嘧啶-5-基)甲基磺酸丙酯(700mg，2.46mmol)和(4-甲氧基苯基)甲酰胺(1.04g，7.4mmol)溶于二氯甲烷(30mL)中，然后加入三乙胺(749mg，7.4mmol)，置换反应体系为氮气体系，反应液在55℃下搅拌过夜。
LCMS监测显示原料消失后，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-(4-甲氧基苄基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(710mg)。
MS(ESI)M/Z:290.1[M+H] +.
步骤E:在室温下，将4-氯-7-(4-甲氧基苄基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(710mg，2.46mmol)溶于三氟乙酸(3mL)和三氟甲磺酸(0.3mL)中，反应液在70℃下搅拌3小时。
LCMS监测显示原料消失后，减压浓缩。混合液用饱和碳酸氢钠溶液调节pH＝8-9，然后用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(280mg)。
MS(ESI)M/Z:170.0[M+H] +.
步骤F:在封管中，将4-氯-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(460mg，2.72mmol)和(3aS,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(763.5mg，4.08mmol，参考文献J.Org.Chem.2004,69,3993-3996方法制备，其全文内容通过引用结合到本申请中)溶于甲苯(2mL)。将氰基亚甲基三正丁基膦(1.31g，5.44mmol)溶于甲苯(2mL)中，在氮气氛围下滴加上述液体，反应液在110℃下搅拌18小时。
LCMS监测显示原料消失后，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯 基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(300mg)。
MS(ESI)M/Z:336.2[M+H] +.
步骤G:在室温下，将4-氯-7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(300mg，0.895mmol)溶于乙醇(20mL)和氨水(100mL)混合液中，反应液在160℃高压反应釜中搅拌60小时。
LCMS监测显示大部分是产物后，减压浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(120mg)。
MS(ESI)M/Z:317.0[M+H] +.
步骤H:在一个三口瓶中加入化合物7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5-甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(38mg，0.11mmol)的四氢呋喃(4mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(1.1mmol)的四氢呋喃溶液(0.5M，2.2mL)，滴加完毕后，反应液被加热到55℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(233mg，1.1mmol)的水溶液(1mL)。反应液在室温下搅拌30分钟后，加入3-溴-7-碘喹啉-2-胺(38.5mg，0.11mmol)和PdCl2(dppf)(8mg，0.01mmol)的四氢呋喃(1mL)溶液。最后所得的反应混合液被加热到55℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到7-(2-((3aR，4S，6R，6aS)-6-(4-氨基-5-甲基-5H-吡咯[2,3-d]嘧啶-7(6H)-基)-2,2-二甲基四氢-3aH-环戊烷[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(30mg)。
MS(ESI)M/Z:539.0[M+H] +.
步骤I:在室温下，将7-(2-((3aR，4S，6R，6aS)-6-(4-氨基-5-甲基-5H-吡咯[2,3-d]嘧啶-7(6H)-基)-2,2-二甲基四氢-3aH-环戊烷[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(30mg，0.055mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于5mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8～9。所得溶液经制备型高效液相色谱纯化得到最终产物(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5-甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(15mg)。
MS(ESI)M/Z:499.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.79(d,J＝6.2Hz,1H),7.57(d,J＝8.3Hz,1H),7.30(d,J＝3.5Hz,1H),7.13–7.06(m,1H),6.56(s,2H),5.89(d,J＝22.1Hz,2H),4.86(d,J＝5.6Hz,1H),4.62(d,J＝5.8Hz,1H),4.40(dd,J＝6.6,4.8Hz,1H),4.18-4.10(m,1H),3.92-3.82(m,1H),3.56–3.49(m,1H),3.23–3.13(m,1H),3.08–2.98(m,1H),2.78–2.62(m,2H),1.95–1.80(m,2H),1.78–1.68(m,1H),1.62–1.50(m,1H),1.21–1.16(m,1H),1.15–1.08(m,3H).
实施例35
(1S，2R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇
Figure PCTCN2022106401-appb-000134
反应路线:
Figure PCTCN2022106401-appb-000135
操作步骤：
步骤A:在一个三口瓶中加入化合物4-氯-7-((3aS，4R，6aR)-2,2-二甲基-6-乙烯基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶(75mg，0.23mmol)的四氢呋喃(1mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(2.35mmol)的四氢呋喃溶液(0.5M，4.7mL)，滴加完毕后，反应液被加热到55℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(498mg，2.35mmol)的水溶液(2mL)。反应液在室温下搅拌30分钟后，加入3-溴-7-碘喹啉-2-胺(80.5mg，0.23mmol)和PdCl2(dppf)(14.6mg，0.02mmol)的四氢呋喃(2mL)溶液。最后所得的反应混合液被加热到55℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到3-溴-7-(2-((3aS，6aR)-4-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)喹啉-2-胺(120mg)。
MS(ESI)M/Z:542.0[M+H] +.
步骤B:在室温下，将3-溴-7-(2-((3aS，6aR)-4-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧 啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)喹啉-2-胺(120mg，0.22mmol)溶于乙醇(5mL)和氨水(30mL)中。反应液在140℃高压反应釜中搅拌2天。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS，6aR)-2,2-二甲基-6-乙烯基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(30mg)。
MS(ESI)M/Z:523.3[M+H] +.
步骤C：在室温下，将7-((3aS，6aR)-2,2-二甲基-6-乙烯基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(30mg，0.057mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S，2R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇(3.3mg)。
MS(ESI)M/Z:483.0[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ8.32(s,1H),7.79(s,1H),7.57(d,J＝8.2Hz,1H),7.31(s,1H),7.11(dd,J＝8.2,1.6Hz,1H),6.56(s,2H),5.93(s,2H),5.28(d,J＝1.4Hz,1H),4.82(s,1H),4.74(dd,J＝6.2,4.8Hz,2H),4.29(t,J＝5.8Hz,1H),3.88(dd,J＝10.6,5.8Hz,1H),3.22(dd,J＝16.8,9.0Hz,1H),2.98(dd,J＝18.8,9.2Hz,1H),2.93–2.80(m,2H),2.57(t,J＝9.0Hz,2H),2.48–2.38(m,2H).
实施例36
(1S,2R,3S)-3-(2-(2-氨基-3-氯喹啉-7-基)乙基)-5-(4-甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-醇
Figure PCTCN2022106401-appb-000136
制备方法参考实施例34，最后得到目标产物(1S,2R,3S)-3-(2-(2-氨基-3-氯喹啉-7-基)乙基)-5-(4-甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-醇。
MS(ESI)M/Z:440.4[M+H] +.
1H NMR(400MHz,CDCl 3)δ8.27(s,1H),7.95(s,1H),7.54–7.47(m,2H),7.18(dd,J＝8.2,1.5Hz,1H),5.38(s,2H),4.12–4.04(m,1H),3.91–3.83(m,2H),3.78–3.70(m,1H),3.51(dd,J＝9.6Hz,1H),3.00(t,J＝7.6Hz,2H),2.85(t,J＝7.7Hz,2H),2.31–2.22(m,4H),2.11–1.94(m,4H),1.75–1.70(m,1H),1.46–1.35(m,1H).
实施例37
(1S,2R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇
Figure PCTCN2022106401-appb-000137
制备方法参考实施例35，最后得到目标产物(1S,2R)-3-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇。
MS(ESI)M/Z:481.0[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.27(s,1H),7.87(s,1H),7.62–7.54(m,2H),7.50(d,J＝8.4Hz,1H),7.11–6.97(m,2H),5.99(s,1H),5.09(d,J＝6.0Hz,1H),4.24–4.18(m,1H),3.71–3.67(m,1H),3.56–3.52(m,2H),2.89–2.85(m,2H).
实施例38
(1S，2R)-3-(2-(2-氨基-3-氯喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇
Figure PCTCN2022106401-appb-000138
反应路线:
Figure PCTCN2022106401-appb-000139
操作步骤：
步骤A:在一个三口瓶中加入化合物4-氯-7-((3aS，4R，6aR)-2,2-二甲基-6-乙烯基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶(110mg，0.34mmol)的四氢呋喃(1mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(3.44mmol)的四氢呋喃溶液(0.5M，6.89mL)，滴加完毕后，反应液被加热到55℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(729mg，3.44mmol)的水溶液(2mL)。反应液在室温下搅拌30分钟后，加入7-溴-3-氯喹啉-2-胺(88mg，0.34mmol)和PdCl 2(dppf)(21.9mg，0.03mmol)的四氢呋喃(2mL)溶液。最后所得的反应混合液被加热到55℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品。粗产品用硅胶板层析纯化得到3-氯-7-(2-((3aS，6aR)-4-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)喹啉-2-胺(100mg)。
MS(ESI)M/Z:498.0[M+H] +.
步骤B:在室温下，将3-氯-7-(2-((3aS，6aR)-4-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)喹啉-2-胺(60mg，0.12mmol)溶于乙醇(5mL)和氨水(30mL)中。反应液在140℃高压反应釜中搅拌2天。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-(2-((3aS，6aR)-4-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)-3-氯喹啉-2-胺(20mg)。
MS(ESI)M/Z:479.0[M+H] +.
步骤C：在室温下，将7-(2-((3aS，6aR)-4-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二氧基-6-基)乙基)-3-氯喹啉-2-胺(20 mg，0.042mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S，2R)-3-(2-(2-氨基-3-氯喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊-3-烯-1,2-二醇(2.7mg)。
MS(ESI)M/Z:439.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.07(s,1H),7.81(s,1H),7.57(d,J＝8.2Hz,1H),7.37(s,1H),7.23-7.15(m,1H),5.35(s,1H),4.58(s,1H),4.42(d,J＝5.8Hz,1H),4.04–3.96(m,1H),3.26–3.20(m,2H),3.02–2.95(m,2H),2.69–2.53(m,4H).
实施例39
(1R，2S，3R，5R)-3-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-5-((E)-2-(2-氨基喹唑啉-7-基)乙烯基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000140
反应路线:
Figure PCTCN2022106401-appb-000141
操作步骤：
步骤A:在室温下，将7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺(100mg，0.33mmol)，7-溴喹唑啉-2-胺(81.2mg，0.33mmol)和四乙基氯化铵(60.15mg，0.363mmol)溶于N，N- 二甲基甲酰胺(1.5mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(15mg，0.066mmol)，N，N-二异丙基乙胺(85mg，0.66mmol)。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aR，4R，6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)喹唑啉-2-胺(130mg)。
MS(ESI)M/Z:446.2[M+H] +.
步骤B：在室温下，将7-((E)-2-((3aR，4R，6R，6aS)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)喹唑啉-2-胺(130mg，0.29mmol)溶于甲醇(2mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(5mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1R，2S，3R，5R)-3-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-5-((E)-2-(2-氨基喹唑啉-7-基)乙烯基)环戊烷-1,2-二醇(7.7mg)。
MS(ESI)M/Z:406.2[M+H] +.
1H NMR(400MHz,DMSO-d 6)δ9.00(s,1H),7.80(s,1H),7.70(d,J＝8.0Hz,1H),7.39(d,J＝8.8Hz,1H),7.31(s,1H),6.76(s,2H),6.56(s,2H),5.94(s,2H),4.83(d,J＝5.2Hz,1H),4.69(d,J＝5.6Hz,1H),4.22(dd,J＝16.0,8.8Hz,1H),3.97(dd,J＝11.2,5.6Hz,1H),3.73(dd,J＝11.6,5.8Hz,1H),3.52(t,J＝8.8Hz,2H),2.78–2.68(m,2H),2.65–2.61(m,1H),1.98-1.95(m,1H),1.58-1.50(m,1H).
实施例40
(1S，2R，3S，5R)-3-(2-(2-氨基-3-氟喹啉-7-基)乙基)-5-(4-氨基-9H-嘧啶[4,5-b]吲哚-9-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000142
反应路线:
Figure PCTCN2022106401-appb-000143
操作步骤：
步骤A:在封管中，将4-氯-9H-嘧啶并[4,5-b]吲哚(660mg，3.24mmol)和(3aS,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(890mg，4.88mmol，参考文献J.Org.Chem.2004,69,3993-3996方法制备，其全文内容通过引用结合到本申请中)溶于干燥甲苯(5mL)。将氰基亚甲基三正丁基膦(1.56g，6.48mmol)溶于甲苯(3mL)中，在氮气氛围下滴加上述液体，反应液在110℃下搅拌18小时。
LCMS监测显示原料消失后，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-9-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚(379mg)。
MS(ESI)M/Z:370.0[M+H] +.
1H NMR(400MHz,DMSO)δ8.87(s,1H),8.38(d,J＝8.0Hz,1H),7.97(d,J＝8.4Hz,1H),7.73(t,J＝7.8Hz,1H),7.51(t,J＝7.6Hz,1H),6.07-5.98(m,1H),5.45–5.38(m,1H),5.30(dd,J＝7.2,5.8Hz,1H),5.19(d,J＝17.2Hz,1H),5.11(d,J＝10.4Hz,1H),4.76(t,J＝7.0Hz,1H),2.88-2.76(m,1H),2.71(dd,J＝24.4,12.0Hz,1H),2.34–2.25(m,1H),1.53(s,3H),1.22(s,3H).
步骤B:在封管中，将4-氯-9-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚(379mg，1.02mmol)溶于乙醇(4mL)和氨水(10mL)混合液中，反应液在120℃封管中搅拌18小时。
LCMS监测显示大部分是产物后，减压浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS，4R，6R，6aR)-9-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚-4-胺(179mg)。
MS(ESI)M/Z:351.2[M+H] +.
步骤C:在一个三口瓶中加入化合物7-((3aS，4R，6R，6aR)-9-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚-4-胺(60mg，0.17mmol)的四氢呋喃(1mL)溶液，然后体系置换为氮气氛围。室温下，向反应液中缓慢滴加9-BBN(1.19mmol)的四氢呋喃溶液(0.5M，2.39mL)，滴加完毕后，反应液被加热到55℃，在此温度下搅拌1小时。反应体系被移出油浴冷却到室温，然后向反应混合液中加入磷酸钾(252mg，1.19mmol)的水溶液(2mL)。反应液在室温下搅拌30分钟后，加入7-溴-3-氟喹啉-2-胺(41mg，0.17mmol，参考实施例31的7-溴-3-氯喹啉-2-胺的合成方法制备)和PdCl 2(dppf)(14.6mg，0.02mmol)的四氢呋喃(2mL)溶液。最后所得的反应混合液被加热到55℃，搅拌过夜。
TLC监测显示原料消失后，反应液被减压浓缩得到粗产品纯化得到9-((3aS，4R，6S，6aR)-6-(2-(2-氨基-3-氟喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚-4-胺(60mg)。
MS(ESI)M/Z:513.4[M+H] +.
步骤D:在室温下，将9-((3aS，4R，6S，6aR)-6-(2-(2-氨基-3-氟喹啉-7-基)乙基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-9H-嘧啶[4,5-b]吲哚-4-胺(60mg，0.12mmol)溶于甲醇(2mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(2mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S，2R，3S，5R)-3-(2-(2-氨基-3-氟喹啉-7-基)乙基)-5-(4-氨基-9H-嘧啶[4,5-b]吲哚-9-基)环戊烷-1,2-二醇(28mg)。
MS(ESI)M/Z:473.2[M+H] +.
1H NMR(400MHz,MeOD)δ8.29(s,1H),8.13(d,J＝7.6Hz,1H),7.69–7.56(m,3H),7.49–7.41(m,2H),7.32(t,J＝7.2Hz,1H),7.22(d,J＝8.2Hz,1H),5.13(dd,J＝18.0,8.2Hz,1H),4.97(t,J＝6.8Hz,1H),4.12(t,J＝5.8Hz,1H),2.95–2.80(m,2H),2.32–2.07(m,4H),1.97–1.90(m,1H).
实施例41
(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,5-二甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000144
反应路线:
Figure PCTCN2022106401-appb-000145
操作步骤：
步骤A:在一个三瓶中加入(3aR，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(550mg，1.72mmol)的N，N-二甲基甲酰胺(10mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后分批加入叔丁醇钾粉末(579mg，5.16mmol)。加入完毕后，反应体系继续在0℃下搅拌1小时。然后在氮气氛围下滴加碘甲烷(732mg，5.16mmol)，反应体系在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中缓慢加入水(10mL)淬灭。然后用乙酸乙酯萃取，(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-5,7-二氢-6H-吡咯[2,3-d]嘧啶-6-酮(560mg)。
MS(ESI)M/Z:348.2[M+H] +.
步骤B:在一个三口瓶中加入4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-5,7-二氢-6H-吡咯[2,3-d]嘧啶-6-酮(460mg，1.33mmol)的四氢呋喃(10mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后逐滴滴加四氢铝锂(2.66mmol)的四氢呋喃溶液(1M，2.66mL)。滴加完毕后，反应体系继续在0℃下搅拌1小时。
TLC监测显示原料消失后，向反应液中缓慢加入十水合硫酸钠淬灭直至没有气泡冒出。然后加入四氢呋喃搅拌30分钟，过滤反应液。滤液被浓缩得到油状粗产物，用硅胶柱层析纯化得到4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-6-醇(370mg)。
MS(ESI)M/Z:350.2[M+H] +.
步骤C:在一个三口瓶中加入4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-6-醇(370mg，1.06mmol)的二氯甲烷(20mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后三氟乙酸(2.3mL)和三乙基硅烷(2.3mL)。加入完毕后，反应体系室温下搅拌1小时。
LCMS监测显示原料消失后，向反应液中缓慢加入饱和碳酸氢钠溶液调节pH＝9-10。然后用二氯甲烷萃取(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(20mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(330mg)。
MS(ESI)M/Z:334.2[M+H] +.
步骤D:在单口瓶中，将4-氯-7-(2,4-二甲氧基苄基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(330mg，1.01mmol)溶于三氟乙酸(15mL)中，反应液在70℃下搅拌过夜。
LCMS监测显示原料消失后，减压浓缩。混合液用饱和碳酸氢钠溶液调节pH＝8-9，然后用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(180mg)。
MS(ESI)M/Z:184.2[M+H] +.
步骤E:在封管中，将4-氯-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(80mg，0.44mmol)和(3aS,4S,6R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊二烯并[d][1,3]二氧杂环-4-醇(123.4mg，0.66mmol，参考文献J.Org.Chem.2004,69,3993-3996方法制备，其全文内容通过引用结合到本申请中)溶于干燥甲苯(1mL)。将氰基亚甲基三正丁基膦(318mg，1.32mmol)溶于甲苯(0.5mL)中，在氮气氛围下滴加上述液体，反应液在110℃下搅拌18小时。
LCMS监测显示原料消失后，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到4-氯-7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(140mg)。
MS(ESI)M/Z:350.2[M+H] +.
步骤F:在室温下，将4-氯-7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶(140mg，0.4mmol)溶于乙醇(50mL)和氨水(150mL)混合液中，反应液在160℃高压反应釜中搅拌60小时。
LCMS监测显示大部分是产物后，减压浓缩。所得残余物用硅胶柱层析纯化得到7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,5-二 甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺)(120mg)。
MS(ESI)M/Z:331.2[M+H +].
步骤G:取一个干燥的三口瓶，将体系置换为氮气氛围，加入7-((3aS，4R，6R，6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,5-二甲基-6,7-二氢-5H-吡咯[2,3-d]嘧啶-4-胺)(60mg，0.182mmol)的四氢呋喃(2mL)溶液，在冰浴下滴加9-BBN(1.274mmol)的四氢呋喃溶液(0.5M，2.55mL)，反应体系在55℃油浴下加热1小时，然后将反应液冷却至室温。将磷酸钾(370mg，1.274mmol)溶于四氢呋喃(1mL)中。向反应体系中滴加上述溶液，加入完毕后，反应体系室温下搅拌20分钟。然后加入3-溴-7-碘喹啉-2-胺(63.25mg，0.182mmol)和1,1'-双二苯基膦二茂铁二氯化钯(13.3mg，0.0182mmol)，反应液在55℃油浴中搅拌过夜。
LCMS监测显示原料消失后，减压浓缩。混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(20mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-(2-((3aR，4S，6R，6aS)-6-(4-氨基-5,5-二甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(30mg)。
MS(ESI)M/Z:553.2[M+H] +.
步骤H:在室温下，将7-(2-((3aR，4S，6R，6aS)-6-(4-氨基-5,5-二甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙基)-3-溴喹啉-2-胺(30mg，0.054mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(1mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，向反应液中加入1M的氢氧化钠溶液调节pH值约等于8～9。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残留物经制备型高效液相色谱纯化得到最终产物(1S，2R，3S，5R)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-5,5-二甲基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(4.2mg)。
MS(ESI)M/Z:513.3[M+H] +.
1H NMR(400MHz,MeOD)δ8.26(s,1H),7.83(s,1H),7.56(d,J＝8.2Hz,1H),7.40(s,1H),7.18(dd,J＝8.2,1.6Hz,1H),4.29–4.22(m,1H),4.00(dd,J＝8.2,6.2Hz,1H),3.71(dd,J＝6.0,4.2Hz,1H),3.54–3.33(m,2H),2.85–2.75(m,2H),2.12–2.05(m,1H),2.02–1.89(m,2H),1.73–1.70(m,1H),1.37(s,6H),1.34–1.25(m,1H).
实施例42
(1S,2R,3S)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-yl)-3-甲基环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000146
制备方法参考实施例5，最后得到目标产物(1S,2R,3S)-3-(2-(2-氨基-3-溴喹啉-7-基)乙基)-5-(4-氨基-6,7-二氢环戊烷[4,5]吡咯[2,3-d]嘧啶-8(5H)-yl)-3-甲基环戊烷-1,2-二醇。
MS(ESI)M/Z:537.6[M+H] +.
1H NMR(400MHz,MeOD)δ8.26(s,1H),7.98(s,1H),7.56(d,J＝8.2Hz,1H),7.41(s,1H),7.19(dd,J＝8.2,1.6Hz,1H),4.99–4.92(m,1H),4.63–4.56(m,1H),3.90(d,J＝6.2Hz,1H),2.97–2.89(m,4H),2.89–2.71(m,2H),2.54–2.48(m,2H),2.08–1.83(m,4H),1.22(s,3H).
实施例43
(1R,2S,3R,5R)-3-((E)-2-(2-氨基-3-(三氟甲基)喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000147
反应路线:
Figure PCTCN2022106401-appb-000148
操作步骤：
步骤A:在室温下，将7-溴-3-(三氟甲基)喹啉(950mg，3.44mmol)溶于二氯甲烷(25mL)中。在0℃下缓慢加入间氯过苯甲酸(2.37g，13.7mmol)，反应液在室温下搅拌12小时。
LCMS监测显示原料消失后，0℃下向反应液中加入水(50mL)淬灭，然后缓慢加入氯化铵的饱和水溶液调节pH至7，减压浓缩。混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-溴-3-(三氟甲基)喹啉1-氧化物(650mg)。
MS(ESI)M/Z:292.0[M+H] +.
步骤B:在室温下，将7-溴-3-(三氟甲基)喹啉1-氧化物(650mg，2.24mmol)，溶于三氯甲烷(15mL)中。然后在0℃下缓慢加入三氯氧磷(15mL)，反应体系抽真空换氮气多次。反应液在70℃油浴下搅拌3小时。
LCMS监测显示原料消失后，减压浓缩。加入二氯甲烷(30mL)，然后用饱和碳酸氢钠溶液调节pH至7，混合液用乙酸乙酯(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(30mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-溴-2-氯-3-(三氟甲基)喹啉(560mg)。
MS(ESI)M/Z:310.0[M+H] +.
步骤C:在室温下，将7-溴-2-氯-3-(三氟甲基)喹啉(560mg，1.8mmol)溶于乙醇(10mL)和氨水(20mL)中。随后反应液置于焖罐中在100℃下搅拌20小时。
LCMS监测显示原料消失后，减压浓缩，混合液用二氯甲烷(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-溴-3-(三氟甲基)喹啉-2-胺(400mg)。
MS(ESI)M/Z:291.4[M+H] +.
步骤D:在室温下，将8-((3aS,4R,6aR)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧醇-4-基)-5,6,7,8-四氢环戊烷[4,5]吡咯[2,3-d]嘧啶-4-胺(80mg，0.26mmol)，7-溴-3-(三氟甲基)喹啉-2-胺(75mg，0.26mmol)和四乙基氯化铵(65mg，0.39mmol)溶于N，N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(6.7mg，0.03mmol)，N,N-二异丙基乙胺(0.09mL，0.54mmol)。反应液在100℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(20mL)淬灭，减压浓缩。混合液用乙酸乙酯(20mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化得到7-((E)-2-((3aS，4R，6R，6aR)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)-3-(三氟甲基)喹啉-2-胺(80mg)。
MS(ESI)M/Z:512.6[M+H] +.
步骤E：在室温下，将7-((E)-2-((3aS，4R，6R，6aR)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二氧基-4-基)乙烯基)-3-(三氟甲基)喹啉-2-胺(80mg，0.156mmol)溶于甲醇(1mL)中。随后，向上述溶液中加入4M的盐酸-甲醇溶液(3 mL)。反应液在室温下搅拌2小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产物。粗产物被溶于2mL甲醇，然后加入7M氨甲醇溶液调节pH值约等于8～9。所得溶液经制备型高效液相色谱纯化得到最终产物(1R，2S，3R，5R)-3-((E)-2-(2-氨基-3-(三氟甲基)喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(4mg)。
MS(ESI)M/Z:472.6[M+H] +.
1H NMR(400MHz,MeOD)δ8.35(s,1H),7.76(s,1H),7.72(d,J＝8.3Hz,1H),7.52-7.46(m,2H),6.67–6.63(m,1H),6.57–6.51(m,1H),4.38-4.24(m,1H),4.16–4.12(m,1H),3.92–3.88(m,1H),3.76-3.61(m,2H),2.93-2.73(m,3H),2.21-2.07(m,1H),1.75-1.59(m,1H).
实施例44
(1R,2S,3S,5R)-3-(2-(2-氨基-3-(三氟甲基)喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000149
制备方法参考实施例43，最后得到目标产物(1R,2S,3S,5R)-3-(2-(2-氨基-3-(三氟甲基)喹啉-7-基)乙基)-5-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇。
MS(ESI)M/Z:474.6[M+H] +.
1H NMR(400MHz,MeOD)δ8.36(s,1H),7.82(s,1H),7.72(d,J＝8.2Hz,1H),7.44(s,1H),7.24(d,J＝8.2Hz,1H),4.25-4.19(m,1H),4.10-3.99(m,1H),3.79-3.55(m,3H),2.96–2.75(m,4H),2.15-2.02(m,1H),2.02-1.87(m,2H),1.83–1.65(m,1H),1.42-1.26(m,1H).
实施例45
2-氨基-7-((E)-2-((1R,2R,3S,4R)-4-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,3-二羟基环戊基)乙烯基)喹啉-3-甲腈
Figure PCTCN2022106401-appb-000150
反应路线:
Figure PCTCN2022106401-appb-000151
操作步骤：
步骤A：将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯[d][1,3]二氧戊环-4-基)乙烯基)-3-溴喹啉-2-胺(参照实施例14步骤B产物的合成方法制备，100.0mg，0.19mmol)，氰化亚铜(70mg，0.77mmol)，三(二亚苄基丙酮)二钯(20mg，0.022mmol)和1,1'-双(二苯基膦)二茂铁(43mg，0.077mmol)加入1，4-二氧六环(6mL)中，氮气置换3次，反应液升温至100℃，搅拌过夜。
TLC监测显示原料消失后，冷却，直接减压浓缩。所得残余物用硅胶柱层析纯化得到2-氨基-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7–基)-2,2-二甲基四氢-4H-环戊二烯[d][1,3]二氧戊环-4-基)乙烯基)喹啉-3-甲腈(50mg)。
MS(ESI)M/Z:470.2[M+H] +.
步骤B：将2-氨基-7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7–基)-2,2-二甲基四氢-4H-环戊二烯[d][1,3]二氧戊环-4-基)乙烯基)喹啉-3-甲腈(50mg，0.106mmol)溶于三氟乙酸(2mL)和水(1mL)中，室温搅拌2小时。
LCMS监测原料反应完，反应液加入乙腈稀释，直接制备提纯得到2-氨基-7-((E)-2-((1R,2R,3S,4R)-4-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,3-二羟基环戊基)乙烯基)喹啉-3-甲腈(4.06mg)。
MS(ESI)M/Z:430.1[M+H] +.
1H NMR(400MHz,CD 3OD)δ8.77(s,1H),8.13(s,1H),7.83(d,J＝8.5Hz,1H),7.68–7.58(m,2H),6.69(d,J＝3.4Hz,2H),4.57–4.48(m,1H),4.20–4.16(m,1H),3.99–3.86(m,3H),3.02–2.98(m,2H),2.85–2.79(m,1H),2.24–2.09(m,1H).
实施例46
(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-(甲基磺酰基)喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇
Figure PCTCN2022106401-appb-000152
反应路线:
Figure PCTCN2022106401-appb-000153
操作步骤：
步骤A：将7-((E)-2-((3aR,4R,6R,6aS)-6-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)-2,2-二甲基四氢-4H-环戊二烯[d][1,3]二氧戊环-4-基)乙烯基)-3-溴喹啉-2-胺(参照实施例14步骤B产物的合成方法制备，150.0mg，0.29mmol)，甲烷亚磺酸钠(138mg，1.35mmol)，脯氨酸(34mg，0.3mmol)，碘化亚铜(36mg，0.19mmol)和碳酸铯(66mg，0.2mmol)加入二甲亚砜(5mL)中，氮气保护下，升温至100℃，搅拌2.5小时。
TLC监测显示原料大部分消失，冷却至室温，向反应体系中加水(20mL)。混合液用乙酸乙酯(30mL×2)萃取，合并有机相，有机相先用饱和食盐水(30mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得粗品直接用于下一步。
MS(ESI)M/Z:523.1[M+H] +.
步骤B：将上步粗品溶于甲醇(5mL)，向其中加入4M盐酸水溶液(2mL)，反应液室温搅拌2小时。
TLC监测原料反应完，向反应体系中入饱和碳酸氢钠水溶液(20mL)淬灭，混合液用二氯甲烷(30mL×3次)萃取，合并有机相，有机相先用饱和食盐水(10mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。粗品制备提纯得到(1S,2R,3R,5R)-3-((E)-2-(2-氨基-3-(甲基磺酰基)喹啉-7-基)乙烯基)-5-(4-氨基-5,6-二氢-7H-吡咯并[2,3-d]嘧啶-7-基)环戊烷-1,2-二醇(14.71mg)。
MS(ESI)M/Z:483.1[M+H] +.
1H NMR(400MHz,CD 3OD)δ9.00(s,1H),8.16(s,1H),8.00(d,J＝8.5Hz,1H),7.78-7.69(m,2H),6.84–6.68(m,2H),4.59–4.53(m,1H),4.24–4.20(m,1H),4.02–3.84(m,3H),3.34(s,3H),3.07–2.97(m,2H),2.88–2.84(m,1H),2.20–2.16(m,1H),1.80–1.76(m,1H).
实施例47
(1S,4R,6R,7R)-4-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2-氧杂二环[2.2.1]环戊烷-7-醇
Figure PCTCN2022106401-appb-000154
反应路线:
Figure PCTCN2022106401-appb-000155
操作步骤：
步骤A:在室温下，将(3aR，6aR)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二恶英-4-酮(924mg，6.0mmol)和二苯甲酮(218mg，1.2mmol)溶于甲醇(400mL)中。反应体系用氮气置换保护，然后将反应瓶置于暗室，在汞灯的照射下搅拌2小时。
TLC监测显示原料消失后，将反应液减压浓缩得到粗产品。所得粗产品用硅胶柱层析纯化得到(3aR,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(686mg)。
1H NMR(400MHz,DMSO-d 6)δ4.92(t,J＝4.6Hz,1H),4.63(d,J＝5.4Hz,1H),4.18(d,J＝5.4Hz,1H),3.64–3.52(m,1H),3.46–3.38(m,1H),2.63–2.52(m,2H),2.41–2.34(m,1H),1.31(s,3H),1.26(s,3H).
步骤B:在室温下，将(3aR,6aR)-6-(羟甲基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(5.58g，30mmol)和咪唑(4.08g，60mmol)溶于二氯甲烷(60mL)中。然后将反应液冷却到0℃，向反应液中滴加叔丁基二苯基氯硅烷(12.37g，450mmol)的二氯甲烷溶液。反应液在室温下搅拌16小时。
TLC监测显示原料消失后，向反应液中加入水(300mL)淬灭，混合液用二氯甲烷(60mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化，得到(3aR，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(10.96g)。
1H NMR(400MHz,DMSO-d 6)δ7.63–7.55(m,4H),7.52–7.42(m,6H),4.73–4.60(m,1H),4.21(d,J＝5.4Hz,1H),3.78(dd,J＝10.1,2.9Hz,1H),3.59(dd,J＝10.1,3.6Hz,1H),2.80–2.61(m,1H),2.54–2.52(m,1H),2.49–2.45(m,1H),1.33(s,3H),1.27(s,3H),0.95(s,9H).
步骤C:在一个三口瓶中，加入(3aR,6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(10.6g，25mmol)和三乙基氯硅烷(7.54g，50mmol)的四氢呋喃(50mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至-78℃，然后逐滴滴加双(三甲基硅基)氨基锂(50mmol)的四氢呋喃溶液(1M，50mL)。滴加完毕后，反应体系继续在0℃下搅拌40分钟。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(100mL×3次)萃取，合并有机相，有机相用饱和食盐水(50mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化，得到叔丁基(((3aR，6aR)-2,2-二甲基-6-(三乙基硅基)氧基)-3a，6a-二氢-4H-环戊烷[d][1,3]二恶英-4-基)甲氧基)二苯基硅烷(6.25g)。
步骤D:在一个单口瓶中，加入叔丁基(((3aR，6aR)-2,2-二甲基-6-(三乙基硅基)氧基)-3a，6a-二氢-4H-环戊烷[d][1,3]二恶英-4-基)甲氧基)二苯基硅烷(6.25g，11.6mmol)，醋酸钯(779mg，3.48mmol)和二甲基亚砜(100mL)，然后体系置换为氧气氛围。反应液被加热到60℃，并在此温度下反应过夜。
TLC监测显示原料消失后，将反应液倒入冰水中。反应液用乙酸乙酯(60mL×3次) 萃取，合并有机相，有机相依次用水(60mL×3次)和饱和食盐水(60mL)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化，得到(3aR，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二恶英-4-酮(3.45g，收率70.5％)。
1H NMR(400MHz,CDCl 3)δ7.71–7.60(m,4H),7.48–7.33(m,6H),6.33(t,J＝1.9Hz,1H),4.98(d,J＝5.6Hz,1H),4.69(dd,J＝18.9,1.9Hz,1H),4.54–4.41(m,2H),1.34(d,J＝8.0Hz,6H),1.12–1.05(m,9H).
步骤E:在一个三口瓶中，加入氯化锂(127mg，3mmol)，碘化亚铜(286mg，1.5mmol)和四氢呋喃(100mL)，然后体系置换为氮气氛围。反应液被冷却到0℃并在此温度下搅拌10分钟，然后向反应液中逐滴滴加三甲基氯硅烷(1.956g，18mmol)和(3aR，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-3a，6a-二氢-4H-环戊烷[d][1,3]二恶英-4-酮(6.34g，15mmol)的四氢呋喃(40mL)溶液。滴加完毕后，反应液继续在0℃下搅拌20分钟。然后逐滴滴加乙烯基溴化镁(24mmol)的四氢呋喃溶液(1M，24mL)。所得反应液在0℃下继续搅拌30分钟。
TLC监测显示原料消失后，向反应液中加入饱和氯化铵水溶液淬灭。反应液用乙酸乙酯(50mL×3次)萃取，合并有机相，有机相用饱和食盐水(30mL×3次)洗涤，无水硫酸钠干燥，过滤，减压浓缩。所得混合物用硅胶柱层析纯化，得到(3aR，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(4.15g)。
1H NMR(400MHz,CDCl 3)δ7.76–7.64(m,4H),7.47–7.33(m,6H),6.21–5.86(m,1H),5.29–5.19(m,1H),5.17–5.04(m,1H),4.94–4.70(m,1H),4.34–4.18(m,1H),3.96(d,J＝9.4Hz,1H),3.72(d,J＝9.4Hz,1H),2.46–2.27(m,2H),1.42–1.31(m,6H),1.13–1.03(m,9H).
步骤F:在一个三口瓶中加入(3aR，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英-4-酮(4.15g，9.2mmol)的四氢呋喃(50mL)溶液，然后体系置换为氮气氛围。反应瓶置于冰浴中降温至0℃，然后逐滴滴加四氢铝锂(18.4mmol)的四氢呋喃溶液(1M，18.4mL)。滴加完毕后，反应体系继续在0℃下搅拌2小时。
TLC监测显示原料消失后，向反应液中缓慢加入十水合硫酸钠淬灭直至没有气泡冒出。然后加入四氢呋喃搅拌30分钟，过滤反应液。滤液被浓缩得到油状粗产物，用硅胶柱层析纯化，得到(3aS，4S，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英-4-醇(2.95g)。
1H NMR(400MHz,CDCl 3)δ7.76–7.60(m,4H),7.46–7.30(m,6H),5.94(dd,J＝17.8,11.1Hz,1H),5.18(dd,J＝39.0,14.5Hz,2H),4.60(d,J＝5.4Hz,1H),4.46(t,J＝5.7Hz,1H),4.08–4.00(m,1H),3.82(d,J＝9.3Hz,1H),3.57(d,J＝9.3Hz,1H),1.98(dd,J＝12.2,6.4Hz,1H),1.43(s,3H),1.43–1.38(m,1H),1.36(s,3H),1.03(s,9H).
步骤G：在室温下，将4-氯-6,7-二氢-5H-吡咯并[2,3-d]嘧啶(467mg，3mmol)和(3aS，4S，6aR)-6-(叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二恶英 -4-醇(905mg，2mmol)溶于甲苯(20mL)中，反应体系被置换为氮气氛围。然后将2-(三丁基亚正膦基)乙腈(1.45g，6mmol)的甲苯(2mL)溶液缓慢加入反应体系中。反应液加热到110℃，并在此温度下搅拌16小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产品。粗产品用硅胶柱层析纯化，得到7-(3aS，4R，6R，6aR)-6-((叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-4-氯-6,7-二氢-5H-吡咯[2,3-d]嘧啶(300mg)。
MS(ESI)M/Z:590.1[M+H] +.
步骤H：在室温下，将7-(3aS，4R，6R，6aR)-6-((叔丁基二苯基硅氧基)甲基)-2,2-二甲基-6-乙烯基四氢-4H-环戊烷[d][1,3]二氧基-4-基)-4-氯-6,7-二氢-5H-吡咯[2,3-d]嘧啶(590mg，1mmol)溶于甲醇(3mL)中。向反应混合物加入4N HCl甲醇溶液(2.5mL)。反应液在室温下搅拌16小时。
LCMS监测显示原料消失后，反应液减压浓缩得到粗产品。粗产品用硅胶柱层析纯化，得到(1S，2R，3R，5R)-5-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-3-(羟甲基)-3-乙烯基环戊烷-1,2-二醇(220mg)。
MS(ESI)M/Z:312.1[M+H] +.
步骤I：在室温下，将(1S，2R，3R，5R)-5-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-3-(羟甲基)-3-乙烯基环戊烷-1,2-二醇(218mg，0.7mmol)溶于甲苯(2mL)中，反应体系被置换为氮气氛围。然后将2-(三丁基亚正膦基)乙腈(338mg，1.4mmol)的甲苯(1mL)溶液缓慢加入反应体系中。反应液加热到110℃，并在此温度下搅拌16小时。
LCMS监测显示原料消失后，向反应液中加入水(30mL)淬灭，减压浓缩。混合液用乙酸乙酯(15mL×3次)萃取，合并有机相，有机相先用饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。粗产品用硅胶柱层析纯化，得到(1S，4R，6R，7R)-6-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-4-乙烯基-2-氧杂环[2.2.1]庚烷-7-醇(65mg)。
MS(ESI)M/Z:294.0[M+H] +.
步骤J：在室温下，将(1S，4R，6R，7R)-6-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-4-乙烯基-2-氧杂环[2.2.1]庚烷-7-醇(65mg，0.22mmol)，3-溴-7-碘喹啉-2-胺(92mg，0.264mmol)和四乙基氯化铵(40mg，0.24mmol)溶于N，N-二甲基甲酰胺(2mL)中。反应体系抽真空换氮气多次。在氮气氛围下加入醋酸钯(10mg，0.04mmol)，N，N-二异丙基乙胺(142mg，1.1mmol)。反应液在70℃油浴下搅拌12小时。
LCMS监测显示原料消失后，向反应液中加入水(10mL)淬灭，减压浓缩。混合液用乙酸乙酯(10mL×3次)萃取，合并有机相，有机相先用饱和食盐水(15mL)洗涤，然后用无水硫酸钠干燥，过滤，最后减压浓缩。所得残余物用硅胶柱层析纯化，得到(1S，4R，6R，7R)-4-(E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-6-(4-氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2-氧杂环[2.2.1]庚烷-7-醇(45mg)。
MS(ESI)M/Z:514.1[M+H] +.
步骤K:在室温下，将1S，4R，6R，7R)-4-(E)-2-(2-氨基-3-溴喹啉-7-基)乙烯基)-6-(4- 氯-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2-氧杂环[2.2.1]庚烷-7-醇(45mg，0.29mmol)的乙醇(1mL)溶液加入封管中。随后，在室温下向反应体系中缓慢加入氨水(3mL)。反应液在135℃油浴下搅拌4天。
LCMS监测显示原料消失后，将反应液减压蒸馏浓缩。所得残留物纯化得到最终产物(1S,4R,6R,7R)-4-((E)-2-(2-氨基-3-溴喹啉-7-基)乙烯)-6-(4-氨基-5,6-二氢-7H-吡咯[2,3-d]嘧啶-7-基)-2-氧杂二环[2.2.1]环戊烷-7-醇(2.5mg)。
MS(ESI)M/Z:495.0[M+H] +.
1H NMR(400MHz,MeOD)δ8.26(s,1H),7.88(d,J＝7.0Hz,1H),7.59(d,J＝8.4Hz,1H),7.51(s,1H),7.46(dd,J＝8.5,1.6Hz,1H),6.76(d,J＝16.2Hz,1H),6.63(d,J＝16.2Hz,1H),4.92-4.90(m,1H),4.68(d,J＝6.3Hz,1H)，4.15(dd,J＝9.9,3.9Hz,1H),3.84-3.69(m,2H),2.92-2.88(m,2H),2.12-1.98(m,2H),1.37-1.25(m,2H).
生物学测试评价
测试例1：评价本发明化合物对PRMT5甲基化酶的活性抑制作用
本实验采用TR-FRET(时间分辨荧光共振能量转移)方法测定本发明化合物对PRMT5蛋白甲基化H4组蛋白的短肽底物的抑制活性，并得出本发明化合物对PRMT5甲基化活性抑制的半数抑制浓度(IC 50)。
1.实验材料
PRMT5/MEP50蛋白预混液购自BPS Bioscience公司，组蛋白H4肽购自GL Biochem公司，底物SAM(S-(5’-腺苷基)-L-蛋氨酸氯化物)购自NEB(诺扬生物)公司，TR-FRET reagent购自Perkinelmer公司，缓冲液成分(Bicine(N,N-双(2-羟乙基)甘氨酸)，NaCl，DTT(二硫苏糖醇)，Porcine skin gelatin(明胶)，Tween-20(吐温-20))购自Sigma公司。
2.实验方法
1)采用缓冲液成分以1：3的体积比连续稀释化合物，得到10个不同浓度点的化合物溶液，滴定曲线确定每种化合物溶液的效价。
2)向白色Greiner 384孔板的每个孔中分配100nL化合物溶液，然后分别加入5μL PRMT5/MEP50蛋白预混液和组蛋白H4肽以1：1的体积比混合得到的混合液，室温孵育30分钟。
3)分别加入SAM来引发每个反应，将板密封并置于25℃的恒温箱中，与化合物进行预孵育90分钟。
4)预孵育结束后，每个孔中分别加入10μLTR-FRET reagent检测液，继续孵育60分钟后，在Envision酶标仪(PerkinElmer)上检测TR-FRET信号(激发光320nm或340nm，发射光波长665nm)。
5)使用GraphPad Prism 6软件进行数据分析，计算化合物的IC 50。
本发明化合物对PRMT5甲基化酶抑制作用结果见表1，活性数据分为A、B、C、D四个区间，IC 50≤10nM的化合物用A标识，10nM＜IC 50≤100nM的化合物用B标识，100nM＜IC 50≤500nM的化合物用C标识，500nM＜IC 50＜1000nM用D标识。
表1本发明化合物对PRMT5甲基化酶活性的抑制结果
Figure PCTCN2022106401-appb-000156
结论：从表1中可以看出，本发明化合物对PRMT5甲基化酶具有较好的抑制作用。
测试例2：评价本发明化合物对人胰腺癌细胞株MIA PaCa-2的增殖抑制作用
本实验通过采用化学发光法测定细胞内ATP(腺嘌呤核苷三磷酸)含量来检测本发明化合物对人胰腺癌细胞株MIA PaCa-2的增殖抑制作用，并得出本发明化合物对人胰腺癌细胞株MIA PaCa-2的抑制增殖的半数抑制浓度IC 50。
1.实验材料
DMEM培养基、胎牛血清(FBS)、100×青链霉素混合溶液(Pen/Strep)、GlutaMAX-I Supplement购自GIBCO公司。Cell Titer-Glo发光法细胞活力检测试剂购自Promega公司。人胰腺癌细胞株MIA PaCa-2购自ATCC。
2.实验方法
1)采用加有青链霉素混合溶液的含FBS的DMEM培养基培养人胰腺癌细胞株MIA PaCa-2，于37℃，5vol％CO 2条件下培养至对数期，用细胞计数仪计数，按照每孔800个细胞的密度将细胞接种于96孔培养板，每孔150μL。置于培养箱(37℃，5vol％CO 2)中孵育过夜。
2)第0天：使用D300e(TECAN)超微量加样仪向培养板细胞中加入750nL梯度稀释的待测化合物(9个浓度，1:3比例稀释)，DMSO(二甲基亚砜)终浓度为0.5w/v％，将培养板置于细胞培养箱中孵育6天(37℃，5vol％CO 2)。空白对照加入每孔750nL的DMSO。
3)第6天：每孔加入100μL Cell Titer-Glo试剂，500rpm振荡2分钟，室温避光静置孵育10分钟稳定发光信号。
4)Envision酶标仪(PerkinElmer)检测发光信号。
5)使用GraphPad Prism 6软件进行数据分析，计算化合物的IC 50。
本发明化合物对人胰腺癌细胞株MIA PaCa-2的抑制增殖作用结果见表2，活性数据分为A、B、C、D四个区间，IC 50≤10nM的化合物用A标识，10nM＜IC 50≤100nM的化合物用B标识，100nM＜IC 50≤500nM的化合物用C标识，500nM＜IC 50＜1000nM用D标识。
表2对人胰腺癌细胞株MIA PaCa-2的抑制结果
Figure PCTCN2022106401-appb-000157
结论：从表2中可以看出，本发明化合物对人胰腺癌细胞株MIA PaCa-2具有较好的抑制作用。
测试例3：本发明化合物对稳定表达在中国仓鼠卵巢细胞的hERG钾离子通道的影响
人心室快速激活延迟整流钾电流(I Kr)主要由心脏hERG钾离子通道介导。对I Kr的抑制是非心脏病类药物引起心室肌动作电位时程延长的最常见机理。动作电位时程的延长将导致临床心电图上出现QT间期延长，这与危险的室性心律失常、扭转性心动过速相关。
本试验采用手动膜片钳的方法评估本发明化合物对稳定表达在体外CHO细胞系的hERG(人类ether-à-go-go相关基因)钾通道(I Kr)电流的浓度-反应关系，进而确定本发明化合物是否具有对hERG钾离子通道的抑制作用。
1.实验材料
细胞外液(ECS)：NaCl，145mM；KCl，4mM；CaCl 2，2mM；MgCl 2，1mM；HEPES(4-羟乙基哌嗪乙磺酸)，10mM；葡萄糖，10mM，用纯水溶解，再调节pH至7.3-7.4。中国仓鼠卵巢细胞(CHO-hERG)来源于Sophion生物科学公司。
2.实验方法
1)将本发明部分化合物、溶媒对照、阳性对照(JNJ-64619178)的母液制剂用ECS混合液稀释成浓度为0.3μM、1μM、3μM、10μM和30μM的工作溶液；ECS混合液是ECS和DMSO的混合溶液，其中ECS比例为0.1％(v/v)；空白对照为DMSO。每个浓度重复测定两个细胞。
2)将处于指数生长期的CHO-hERG细胞收集并重悬在ECS中备用。
3)全细胞膜片钳技术下记录hERG电流，记录温度为室温。膜片钳放大器输出信号通过数模转换以及2.9KHz低通滤波。数据记录用Patchmaster Pro软件采集。
4)将hERG电流值转录到Excel表格中，计算化合物对hERG的IC 50。
本发明化合物和JNJ-64619178对hERG钾离子通道的抑制效应见表3。
表3.本发明化合物和JNJ-64619178对hERG电流抑制率
Figure PCTCN2022106401-appb-000158
备注：JNJ-64619178为WO2017032840A1的compound 80，参考专利方法制备。
Figure PCTCN2022106401-appb-000159
结果：从表3可以看出，本发明实施例22和实施例15化合物对hERG的半数抑制浓度(IC 50)值分别为11.34μM和14.81μM，优于对照化合物JNJ-64619178的半数抑制浓度(IC 50)值9.17μM，表明具有心脏安全性更高的潜在优势。
测试例4：本发明化合物对体外人肝微粒体CYP450酶的抑制作用考察评价DDI风险
本实验中建立化合物对人肝微粒体CYP450亚酶CYP3A4的抑制模型，测试化合物设置7个浓度，测定本发明化合物对CYP酶的半数抑制浓度(IC 50)，以表示本发明化合物对CYP酶的抑制作用情况，进而评价化合物的DDI风险。
1.试验方案
1.1试验药品：
本发明部分化合物。
1.2试验材料：
人肝微粒购买自Corning，浓度为0.127mg/mL；磷酸钾缓冲液，100mM；氯化镁，33mM；NADPH(还原型烟酰胺腺嘌呤二核苷酸磷酸)，10mM；终止液，乙腈(含内标100ng/mL甲苯磺丁脲)。
1.3阳性对照、待测化合物和底物：
阳性对照、待测化合物和底物如表4所示。
表4阳性对照、待测化合物和底物
Figure PCTCN2022106401-appb-000160
1.4反应体系：
肝微粒体蛋白浓度，0.127mg/mL；磷酸钾缓冲液，100mM；阳性对照、底物、测试化合物和底物见1.3部分；NADPH，10mM；氯化镁，33mM。
1.5反应过程：
反应体系中加入底物、阳性对照、待测化合物于37℃预孵育10min，然后加入NADPH(还原型烟酰胺腺嘌呤二核苷酸磷酸)辅因子启动反应，于37℃共孵育，其中3A4(美达唑仑)孵育3min，3A4(睾酮)孵育10min，在孵育终点时加入400μM含内标的冷终止液混合终止反应，并于4000rpm离心20min，分离上清液，稀释后进行LC-MS/MS分析。
通过比较测试物对酶的半数抑制浓度(ΙC 50)，评价本发明化合物是否具有DDI(drug-drug interaction，DDI，即药物相互作用)的风险。
测试例5：本发明化合物对体外人肝微粒体CYP450酶的抑制作用考察评价TDI的风险
化合物对CYP酶活性的影响与作用时间相关，一些化合物对酶活性的抑制作用随时间 延长而降低，而有的化合物对酶活性则呈现时间依赖性的不可逆抑制，这对化合物之间的相互作用、化合物的体内代谢及安全性造成影响。本实验中建立化合物对人肝微粒体CYP450亚酶CYP3A4的抑制模型，测试化合物设置7个浓度，测定本发明化合物对CYP酶的时间依赖半数抑制浓度(IC 50)，以评价本发明化合物是否具有TDI(time-dependent inhibition，即时间依赖性抑制)的风险。
1.试验方案
1.1试验药品：
本发明部分化合物。
1.2试验材料：
人肝微粒购买自Corning，浓度为20mg/mL；磷酸钾缓冲液，100mM；氯化镁，300mM；NADP(烟酰胺腺嘌呤二核苷酸磷酸)，65mM；G6P(6-磷酸葡萄糖)，330mM；G6PDH(6-磷酸葡萄糖脱氢酶)，250U/mL；终止液，乙腈(含内标100ng/mL甲苯磺丁脲)。
1.3阳性对照、待测化合物和底物：
阳性对照、待测化合物和底物如表5所示。
表5阳性对照、待测化合物和底物
Figure PCTCN2022106401-appb-000161
1.4反应体系：
肝微粒体蛋白浓度，0.1mg/mL；磷酸钾缓冲液，100mM；阳性对照、底物、测试化合物和底物见1.3部分；NADP，1mM；G6P，5.53mM；G6PDH，1.2U/mL；氯化镁，3.3mM。
1.5反应过程：
反应体系中加入阳性对照、待测化合物于37℃预孵育10min，然后加入NADPH(还原型烟酰胺腺嘌呤二核苷酸磷酸)辅因子启动反应，于37℃共孵育30min，后加入底物混合继续37℃共孵育10min，在孵育终点时加入250μM含内标的冷终止液混合终止反应，并于4000rpm离心20min，分离上清液，稀释后进行LC-MS/MS分析。
通过比较测试物对酶时间依赖的半数抑制浓度(ΙC 50)，评价本发明化合物是否具有TDI(time-dependent inhibition，即时间依赖性抑制)的风险。
2.实验结果
表6.本发明化合物和JNJ-64619178对CYP3A4酶的抑制活性
Figure PCTCN2022106401-appb-000162
结果：从表6可以看出，本发明实施例22和实施例15化合物对CYP3A4酶的IC 50明显高于JNJ-64619178，表明本发明化合物具有显著时间依赖性抑制(time-dependent inhibition，TDI)的风险更低。
测试例6：本发明化合物在比格犬体内药代动力学测定
以比格犬为受试动物，研究本发明化合物经静脉推注、口服或腹腔注射给药后，在特定时间点收集血浆样品，LC-MS/MS检测血浆中化合物浓度，计算PK参数，体现本发明化合物的在犬体内血浆的药代动力学行为。
1.试验方案
1.1试验药品：
本发明部分化合物。
1.2试验动物
比格犬，雄性，供货商为Marshall。
1.3给药
实施例15化合物给药信息：IV(静脉推注)、PO(口服)实验组都为3只犬，IV给药剂量为1mg/kg，给药体积为0.2mL/kg；PO给药剂量为3mg/kg，给药体积为1mL/kg。给药溶媒为5vol％DMSO/10vol％Solutol(增溶剂)/85vol％Saline(生理盐水)。
实施例22化合物给药信息：比格犬：IV、PO实验组都为3只犬，IV给药剂量为1mg/kg，给药体积为0.2mL/kg；PO给药剂量为3mg/kg，给药体积为1mL/kg。给药溶媒为5vol％DMSO/10vol％Solutol/85vol％Saline。
1.4实验器材
离心机购自Eppendorf公司，移液器购自Eppendorf公司。
1.5样品采集
犬给药后，在0.0833(IV)、0.25、0.5、1、2、4、8和24小时，静脉采血各0.5mL，置于EDTA-K2试管中，于4℃、2000g离心10min分离血浆，于-80℃保存。
1.6样品处理
犬血浆样品处理：
1)55μL血浆样品加入200μL乙腈沉淀，涡旋混合后离心15分钟。
2)取处理后上清液用水稀释后通过LC/MS/MS分析待测化合物的浓度。
2.实验结果
药代动力学参数用WinNonlin 6.1计算得到，犬静脉注射、口服和腹腔注射药物的药代 动力学参数见表7。其中，C max表示最大血药浓度，CL表示清除率，Vss表示稳态分布容积，T 1/2表示末端消除半衰期，MRT Inf表示平均驻留时间，AUC表示药时曲线下面积，F表示生物利用度。
表7比格犬静脉注射、口服或腹腔注射本发明部分化合物的药代动力学参数
Figure PCTCN2022106401-appb-000163
注：“/”表示未测定
结果：从表7可以看出，本发明化合物在比格犬体内具有良好的药代动力学性质。
测试例7：本发明化合物在H520异种型移植瘤模型上的体内药效学研究
1.实验动物：BALB/c裸小鼠，雌性，6-8周龄，体重18-22克，由北京维通利华实验动物有限公司提供。
2.实验方法
模型建立：将5×10 6个人肺鳞癌细胞NCI-H520(ATCC，货号HTB-182 TM)接种于BALB/c裸小鼠右侧皮下建立异种移植肿瘤模型。接种后第17天，当平均肿瘤体积达到134mm 3时，挑选肿瘤体积适宜的荷瘤小鼠，根据肿瘤体积进行随机区组化分组，然后开始治疗。第一次和最后一次给药后采集PK血浆以评估药效和暴露量之间的关系。
动物实验分组和给药方案如下表8所示；其中，N表示小鼠数量，PO表示口服给药，QD表示一天一次，4W表示4周。
表8动物实验分组和给药方案
Figure PCTCN2022106401-appb-000164
溶媒：5vol％DMSO+10vol％Solutol+85vol％Saline。
每周两次用游标卡尺测量肿瘤直径。肿瘤体积的计算公式为：V＝0.5a×b 2，a和b分别表示肿瘤的长径和短径。
化合物的抑瘤疗效用肿瘤生长抑制率TGI(％)或相对肿瘤增殖率T/C(％)的评价。
肿瘤生长抑制率TGI(％)＝[(1-(某一天给药组的平均瘤体积－该给药组分组给药时的平均瘤体积))/(某一天溶媒对照组的平均瘤体积－该溶媒对照组分组给药时的平均瘤体积)]×100％。

Claims (23)

  1. 式(I')所示的化合物或其药学上可接受的盐，
    Figure PCTCN2022106401-appb-100001
    其中，R 1选自H或NR 1aR 1b；
    R 1a和R 1b各自独立地选自H或C 1-4烷基；所述的C 1-4烷基可任选被一个或多个R 1c基团所取代，所述R 1c基团为C 3-6环烷基；
    Y选自N或CR 2；
    R 2选自H、卤素、卤素取代的C 1-4烷基、CN或-SO 2-C 1-4烷基；
    L 1选自化学键、-CH 2-、-X-CH 2、-(CH 2) n-CH＝CH-、-CH＝CH-(CH 2) n-、C 3-6环烷基或5-6元杂芳基；
    X选自O、S、NH或CH 2；
    环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基；
    R 3选自羟基或C 1-4烷基；
    m选自0、1、2或者3；
    n选自0或1；
    环B选自
    Figure PCTCN2022106401-appb-100002
    R 4选自NR 4aR 4b或C 1-4烷基；
    R 4a和R 4b各自独立地选自H或C 1-4烷基；
    Figure PCTCN2022106401-appb-100003
    代表单键或双键；
    当
    Figure PCTCN2022106401-appb-100004
    为单键时，R 5和R 6各自独立地选自H、氘、卤素或C 1-4烷基；R 7和R 8各自独立地为H；
    当
    Figure PCTCN2022106401-appb-100005
    为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或C 5-6环烷基。
  2. 根据权利要求1所述的化合物或其药学上可接受的盐，其中所述化合物为式(I)所示的化合物，
    Figure PCTCN2022106401-appb-100006
    其中R 1、R 2、R 3、R 4、R 5、R 6、R 7、R 8、L 1和m如式(I’)所定义。
  3. 根据权利要求1或2所述的化合物或其药学上可接受的盐，其中所述化合物为式(I)所示的化合物，
    Figure PCTCN2022106401-appb-100007
    其中，R 1选自H或NR 1aR 1b；
    R 1a和R 1b各自独立地选自H或C 1-4烷基；所述的C 1-4烷基可任选被一个或多个R 1c基团所取代，所述的R 1c基团为C 3-6环烷基；
    R 2选自H或卤素；
    R 3选自羟基或C 1-4烷基；
    R 4选自NR 4aR 4b或C 1-4烷基；
    R 4a和R 4b各自独立地选自H或C 1-4烷基；
    L 1选自化学键、-CH 2-、-X-CH 2、-(CH 2) n-CH＝CH-、C 3-6环烷基或5-6元杂芳基；
    X选自O、S、NH或CH 2；
    环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基；
    m选自0、1、2或者3；
    n选自0或1；
    Figure PCTCN2022106401-appb-100008
    代表单键或双键；
    当
    Figure PCTCN2022106401-appb-100009
    为单键时，R 5和R 6各自独立地选自H、氘或卤素；R 7和R 8各自独立地为H；
    当
    Figure PCTCN2022106401-appb-100010
    为双键时，R 5和R 8不存在，R 6和R 7与其相连的C原子一起形成苯基或C 5-6元环烷基。
  4. 根据权利要求1-3任一项所述的化合物或其药学上可接受的盐，其中R 1选自H、氨基或
    Figure PCTCN2022106401-appb-100011
  5. 根据权利要求1-4任一项所述的化合物或其药学上可接受的盐，其中R 2选自H、F、Cl、Br、CF 3、CN或-SO 2CH 3。
  6. 根据权利要求1-5中任一项所述的化合物或其药学上可接受的盐，其中L 1选自化学 键、-CH 2-、-O-CH 2、-NH-CH 2、-CH 2-CH 2、-CH＝CH-、-CH 2-CH＝CH-、-CH＝CH-CH 2-、
    Figure PCTCN2022106401-appb-100012
    Figure PCTCN2022106401-appb-100013
  7. 根据权利要求1-6中任一项所述的化合物或其药学上可接受的盐，其中环A选自C 5-6环烷基、C 5-6环烯基或5-10元杂环基，所述的杂环基含有1个或多个选自N或者O的杂原子。
  8. 根据权利要求7所述的化合物或其药学上可接受的盐，其中环A选自
    Figure PCTCN2022106401-appb-100014
    Figure PCTCN2022106401-appb-100015
  9. 根据权利要求1-8中任一项所述的化合物或其药学上可接受的盐，其中R 3选自羟基或甲基；m选自0、1、2或者3。
  10. 根据权利要求8或9所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2022106401-appb-100016
    选自
    Figure PCTCN2022106401-appb-100017
    Figure PCTCN2022106401-appb-100018
  11. 根据权利要求10所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2022106401-appb-100019
    选自
    Figure PCTCN2022106401-appb-100020
    Figure PCTCN2022106401-appb-100021
  12. 根据权利要求1-11中任一项所述的化合物或其药学上可接受的盐，其中R 4选自氨基、甲基或-NHCH 3。
  13. 根据权利要求1-12中任一项所述的化合物或其药学上可接受的盐，其中
    Figure PCTCN2022106401-appb-100022
    代表单键或双键；当
    Figure PCTCN2022106401-appb-100023
    为单键时，R 5和R 6各自独立地选自H、氘、F或甲基；R 7和R 8各自独立地为H；当
    Figure PCTCN2022106401-appb-100024
    为双键时，R 5和R 8不存在，R 6和R 7和与其相连的C原子一起形成苯基或环戊基。
  14. 根据权利要求1-13中任一项所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2022106401-appb-100025
    选自
    Figure PCTCN2022106401-appb-100026
    Figure PCTCN2022106401-appb-100027
  15. 根据权利要求1-14中任一项所述的化合物或其药学上可接受的盐，其中结构单元
    Figure PCTCN2022106401-appb-100028
    为
    Figure PCTCN2022106401-appb-100029
  16. 根据权利要求1-15中任一项所述的化合物或其药学上可接受的盐，其中所述的化合物具有如下式(II)所示的结构：
    Figure PCTCN2022106401-appb-100030
    其中，
    R 9选自H或C 1-4烷基；优选地，R 9选自H或甲基；m选自1或2；
    R 1、R 2、R 3、R 4、R 5、R 6、R 7、R 8和L 1如式(I’)或式(I)所定义。
  17. 根据权利要求16所述的化合物或其药学上可接受的盐，其选自：
    Figure PCTCN2022106401-appb-100031
    其中R 1、R 2、R 4、R 5、R 6、R 7、R 8和R 9如式(II)所定义。
  18. 根据权利要求16所述的化合物或其药学上可接受的盐其选自：
    Figure PCTCN2022106401-appb-100032
    其中R 1、R 2、R 4和R 9如式(II)所定义。
  19. 化合物或其药学上可接受的盐，其选自：
    Figure PCTCN2022106401-appb-100033
    Figure PCTCN2022106401-appb-100034
    Figure PCTCN2022106401-appb-100035
    Figure PCTCN2022106401-appb-100036
  20. 一种药物组合物，其包含权利要求1-19中任一项所述的化合物或其药学上可接受的盐，以及药学上可接受的载体、稀释剂或赋形剂。
  21. 权利要求1-19中任一项所述的化合物或其药学上可接受的盐，或权利要求20所述的药物组合物在制备治疗癌症药物中的应用。
  22. 权利要求1-19中任一项所述的化合物或其药学上可接受的盐，或权利要求20所述的药物组合物在治疗癌症中的用途。
  23. 一种治疗癌症的方法，其包括向有需要的对象施以有效剂量的权利要求1-19中任一项所述的化合物或其药学上可接受的盐，或权利要求20所述的药物组合物。
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