WO2017193924A1 - 螺三元环、螺五元环类肽脱甲酰基酶抑制剂及其在抗菌和抗肿瘤中的应用 - Google Patents
螺三元环、螺五元环类肽脱甲酰基酶抑制剂及其在抗菌和抗肿瘤中的应用 Download PDFInfo
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- GXSZPDMUTYURMZ-AEFFLSMTSA-N CCCC[C@H](CC(NO)=O)C(N(CC1(CC1)C1)[C@@H]1C(Nc1c[n](CCN(C)C)nc1)=O)=O Chemical compound CCCC[C@H](CC(NO)=O)C(N(CC1(CC1)C1)[C@@H]1C(Nc1c[n](CCN(C)C)nc1)=O)=O GXSZPDMUTYURMZ-AEFFLSMTSA-N 0.000 description 1
- LPVILFRMPNHDLT-CABCVRRESA-N CCCC[C@H](CN(C=O)O)C(N(CC1(CC1)C1)[C@@H]1C(Nc1ccncn1)=O)=O Chemical compound CCCC[C@H](CN(C=O)O)C(N(CC1(CC1)C1)[C@@H]1C(Nc1ccncn1)=O)=O LPVILFRMPNHDLT-CABCVRRESA-N 0.000 description 1
- WGVXSSNVGIKLEP-SJORKVTESA-N Cc1cc(NC([C@H](CC2(CC2)C2)N2C([C@H](CC2CCCC2)CN(C=O)O)=O)=O)n[o]1 Chemical compound Cc1cc(NC([C@H](CC2(CC2)C2)N2C([C@H](CC2CCCC2)CN(C=O)O)=O)=O)n[o]1 WGVXSSNVGIKLEP-SJORKVTESA-N 0.000 description 1
- VLIMEPJTWYSGNA-CVEARBPZSA-N Cc1cnc(NC([C@H](CC2(CC2)C2)N2C([C@H](CC2CCCC2)CC(NO)=O)=O)=O)[s]1 Chemical compound Cc1cnc(NC([C@H](CC2(CC2)C2)N2C([C@H](CC2CCCC2)CC(NO)=O)=O)=O)[s]1 VLIMEPJTWYSGNA-CVEARBPZSA-N 0.000 description 1
- YEKKIILUXRITTP-LWKPJOBUSA-N ONC(CC(CC1CCCC1)C(N(CC1(CC1)C1)[C@@H]1C(Nc(cc1)ncc1F)=O)=O)=O Chemical compound ONC(CC(CC1CCCC1)C(N(CC1(CC1)C1)[C@@H]1C(Nc(cc1)ncc1F)=O)=O)=O YEKKIILUXRITTP-LWKPJOBUSA-N 0.000 description 1
- ZMRNIOGLTVRWRE-LRTDYKAYSA-N ONC(CC(CC1CCCC1)C(N(CC1(CC1)C1)[C@H]1c1nc(cccc2)c2[nH]1)=O)=O Chemical compound ONC(CC(CC1CCCC1)C(N(CC1(CC1)C1)[C@H]1c1nc(cccc2)c2[nH]1)=O)=O ZMRNIOGLTVRWRE-LRTDYKAYSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
- C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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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
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/415—1,2-Diazoles
- A61K31/4155—1,2-Diazoles non condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/54—Spiro-condensed
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- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/56—Ring systems containing three or more rings
- C07D209/96—Spiro-condensed ring systems
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
- C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
- C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
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- C—CHEMISTRY; METALLURGY
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
- C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
- C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C—CHEMISTRY; METALLURGY
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- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- the invention belongs to the research field of antibacterial and anticancer drug technology, and relates to a kind of peptide deformylase inhibitor, in particular to a novel spiro three-membered ring and a spiro five-membered cyclic peptide deformylase inhibitor.
- Antibiotics are a series of chemicals that inhibit and kill pathogens at a certain concentration, including metabolites produced by microorganisms, animals and plants, and chemically synthesized or semi-synthetic compounds. Antibiotics not only refer to antibacterial substances, but also anti-tumor, anti-viral, anti-parasitic and other substances are also antibiotics. Antibiotics are an important pillar that enables us to prolong life, live healthier and benefit from modern medicine.
- Chemotherapy is one of the most important means of clinical treatment of malignant tumors.
- tumor cells often become resistant to chemotherapeutic drugs, patients are no longer sensitive to treatment, which ultimately leads to chemotherapy failure.
- EGFR epidermal growth factor receptor
- Iressa and Tarceva have achieved great success in the treatment of non-small cell lung cancer, the drug resistance problem has become increasingly prominent, and half of the drug resistance is derived from the EGFR T790M mutation.
- Hepatocellular carcinoma is one of the most common malignant cancers in the world and is generally highly tolerant to chemotherapeutic drugs. Nearly half of breast cancers are resistant to Herceptin from the start.
- Peptide deformylase is a metalloproteinase that is not only widely found in bacteria but also in Plasmodium falciparum and humans.
- PDF can remove formyl groups on methionine, allowing bacteria to synthesize functionalized proteins.
- the process of removing formyl is a must for bacterial synthesis of proteins. Cheng.
- the main protein synthesis process of human cells does not depend on the process of formyl removal. This difference between bacteria and human synthetic protein makes bacterial PDF a new target for antibacterial drugs.
- the PDF inhibitor prevents the bacteria from undergoing the deformylation step in the process of synthesizing the protein, thereby selectively inhibiting the growth of the bacteria.
- cancer cells such as colorectal cancer, lung cancer, prostate cancer and other human PDF genes are over-expressed, which can affect the energy balance of cells by inhibiting peptide deformylase in cancer cell mitochondria. : thereby depolarizing the mitochondrial membrane of cancer cells, depleting ATP and promoting apoptosis.
- the PDF inhibitor can be developed into a broad-spectrum antibacterial agent with good antibacterial activity and an anticancer drug with anticancer activity.
- PDF inhibitors Metal Binding Group (MBG) and peptidomimetic or non-peptide backbones containing P1, P2, P3 moieties (general structure of the PDF inhibitor of formula (a)).
- MBG Metal Binding Group
- peptidomimetic or non-peptide backbones containing P1, P2, P3 moieties general structure of the PDF inhibitor of formula (a)
- the invention provides a novel structure, a better activity, a less toxic small tricyclic ring, a snail five-membered cyclic peptide deformylase inhibitor, for inhibiting bacteria resistant to existing antibiotics, including Gram Positive bacteria, such as Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, and Gram-negative bacteria that seriously affect human health, such as Moraxella catarrhalis, etc.;
- the spiro-tricyclic and spiro five-membered cyclic peptide deformylase inhibitors selectively inhibit the proliferation of cancer cells such as colorectal cancer, lung cancer, gastric cancer, and liver cancer.
- the spirotricyclic and spiro five-membered cyclic deformylase inhibitors of the present invention belong to antibiotics and comprise four types of structures of the following formulas (1) to (4):
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 1H-pyrazol-3-yl, 5-fluoropyridine 1-oxide-2-yl, 5-(uncle Butyl)isoxazol-3-yl, 6-methyl-N-(4-(pyridin-3-yl)pyrimidin-2-yl)benzene-1-amino-3-yl, 3-fluoropyridine-2 -yl, 5-methylthiazol-2-yl, 3-(pyridin-3-yl)phenyl, N-(pyrimidin-2-yl)benzene-1-amino-3-yl, 4-(pyridine-3 -yl)pyrimidin-2-yl, 4-morpholinephenyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyrimidinyl, 1-methyl-1H-pyrazol-4-yl, 5-iso Oxazolyl, cyclopropyl, 4-methylthiazol
- the spiro three-membered ring, spiro five-membered ring-type peptide deformylase inhibitor is the compound of Examples 1-9, 14-23, 25-41.
- the second category is a first category:
- n 2-4
- R 1 is n-butyl group, cyclopentylmethyl group
- R 2 is an aromatic ring or an aromatic heterocyclic ring.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 2-benzimidazolyl, 1,3,4-oxadioxazole, 1,2,4-oxygen Dioxazole, 1,3,4-triazole.
- the spiro three-membered ring, spiro five-membered ring-type peptide deformylase inhibitor is the compound of Examples 10-13, 24, and 42.
- the third category is a first category:
- n 2-4
- R 1 is n-butyl, cyclopentylmethyl
- R 2 is an aromatic ring or an aromatic heterocyclic ring, a heterocyclic ring, an alkyl group
- R 3 is hydrogen or an alkyl group.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 1H-pyrazol-3-yl, 5-fluoropyridine 1-oxide-2-yl, 5-(uncle Butyl)isoxazol-3-yl, 6-methyl-N-(4-(pyridin-3-yl)pyrimidin-2-yl)benzene-1-amino-3-yl, 3-fluoropyridine-2 -yl, 5-methylthiazol-2-yl, 3-(pyridin-3-yl)phenyl, N-(pyrimidin-2-yl)benzene-1-amino-3-yl, 4-(pyridine-3 -yl)pyrimidin-2-yl, 4-morpholinephenyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyrimidinyl, 1-methyl-1H-pyrazol-4-yl, 5-iso Oxazolyl, cyclopropyl, 4-methylthiazol
- the spiro three-membered ring, spiro five-membered ring-type peptide deformylase inhibitor is a compound of 43-57, 61-74, 77-80.
- n 2-4
- R 1 is n-butyl group, cyclopentylmethyl group
- R 2 is an aromatic ring or an aromatic heterocyclic ring.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 2-benzimidazolyl, 1,3,4-oxadioxazole, 1,2,4-oxygen Dioxazole, 1,3,4-triazole.
- the spirotricyclic, spiro five-membered cyclic peptide deformase inhibitor is a compound of 58-60, 75, 76.
- the molecular weights of the formula (1) to (4) spiro three-membered ring and the spiro five-membered cyclic peptide deformylase inhibitor of the present invention have a molecular weight of 300-600; and are soluble in dichloromethane, acetone, acetonitrile, methanol, Ethanol, N, N-dimethylformamide, dimethyl sulfoxide and other solvents; slightly soluble in ether, water, etc., insoluble in petroleum ether; usually colorless or yellowish powder or foam.
- the present invention also provides the use of the spiro three-membered ring of the formulas (1) to (4) and the spiro five-membered ring-type peptide deformylase inhibitor in inhibiting bacterial peptide deformylase.
- the bacteria include Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, and Moraxella catarrhalis.
- levofloxacin and LBM415 were used as control compounds, and in vitro antibacterial activity screening of forty-six clinical isolates (see Table 1 for strains) was performed on 80 new compounds synthesized (see Table 2 for a list of 80 compounds).
- the present invention also provides the use of the spiro three-membered ring and the spiro five-membered cyclic peptide deformylase inhibitor represented by the formulas (1) to (4) in the preparation of an antitumor drug.
- the tumor refers to colorectal cancer, leukemia, lung cancer, gastric cancer, cervical cancer, breast cancer, prostate cancer, liver cancer, and osteosarcoma.
- the invention adopts the cck-8 method to determine the proliferation inhibition effect of the compound on nine tumor cells such as colorectal cancer, leukemia, lung cancer, gastric cancer, cervical cancer, breast cancer, prostate cancer, liver cancer and osteosarcoma.
- the present invention also provides the use of the spiro three-membered ring and the spiro five-membered cyclic peptide deformylase inhibitor represented by the formulas (1) to (4) for preparing an antiparasitic drug.
- the antiparasitic means that the spiro three-membered ring and the spiro five-membered ring-type peptide deformylase inhibitor inhibit the growth of parasites by inhibiting the parasitic peptide deformylase.
- the present invention also proposes a pharmaceutical composition
- a pharmaceutical composition comprising a spiro three-membered ring, a spiro five-membered ring-type peptide deformylase inhibitor as described above, and a pharmaceutically acceptable carrier.
- the pharmaceutically acceptable carrier refers to a conventional pharmaceutical carrier in the pharmaceutical field, such as a diluent, an excipient, water, etc., a filler such as starch, sucrose, lactose, microcrystalline cellulose, etc.; Cellulose derivatives, alginates, gelatin and polyvinylpyrrolidone; wetting agents such as glycerin; disintegrating agents such as sodium carboxymethyl starch, hydroxypropyl cellulose, croscarmellose, agar, calcium carbonate and carbonic acid Sodium hydride; absorption enhancers such as quaternary ammonium compounds; surfactants such as cetyl alcohol, sodium lauryl sulfate; adsorption carriers such as aged soil and soap clay; lubricants such as talc, calcium and magnesium stearate, fine powder Silica gel and polyethylene glycol.
- flavoring agents, sweeteners and the like can also be added.
- the administration mode of the inhibitor or the pharmaceutical composition includes oral administration, injection, implantation, and the like.
- the inhibitor or the pharmaceutical composition may be administered in the form of a tablet, a liquid or a capsule or the like.
- the novel spiro-containing three-membered ring and spiro five-membered ring-type peptide deformylase inhibitor have antibacterial activity and antitumor activity.
- the mechanism of action of the spiro-like peptide deformylase inhibitor of the present invention is to selectively kill bacteria by inhibiting the activity of the desired peptide deformylase in bacterial protein synthesis without affecting the main protein synthesis process of the human body.
- the energy balance of the cells can be affected: thereby depolarizing the mitochondrial membrane, depleting ATP and promoting apoptosis.
- the inhibitor of the present invention is less susceptible to drug resistance such as commercially available antibiotics due to the novel mechanism of action.
- the peptide-type deformylase inhibitor can be used not only as a new type of antibacterial agent, but also against a variety of antibiotic-resistant Gram-positive strains, such as Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus, Streptococcus pneumoniae; and effective against Gram-negative bacteria such as Moraxella catarrhalis. It can also be used as a new class of anticancer drugs with low micromolar inhibitory activity against various cancer cell strains such as colorectal cancer, lung cancer, gastric cancer and liver cancer.
- the minimum inhibitory concentration MIC of the compound in Example 2 against methicillin-resistant Staphylococcus (MRSA) is 0.125-0.25 ⁇ g/mL
- the MIC of LBM415 is 0.5-1 ⁇ g/mL.
- the compound in the second embodiment of the present invention has been improved four times, and has been significantly improved (see Table 8 in Control Experiment 1); it has been found through experiments that the introduction of the snail five-membered ring increases the activity of the snail five-membered ring inhibitor. Has a significant impact, such as embodiment 77, embodiment 78, embodiment 79, etc.;
- the present invention employs an aromatic amide modification strategy such that some compounds are modified (such as Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, Example 15, Example 16). 19, Example 21, Example 22, Example 23, Example 25, Example 26, Example 27, Example 28, Example 29, Example 30, Example 33, Example 38, etc.
- the activity of the tested strain has been substantially improved.
- Example 4 Example 6, Example 33, Example 38, Example 64, Example 72, Example 73, etc.; and showed very good Gram-negative bacteria Moraxella catarrhalis which seriously affects human health. Good activity, minimum inhibitory concentration as low as 0.06 ⁇ g / mL, as in Example 22, Example 33, etc. (see Control Experiment 3 Table 10).
- the spirotricyclic and spiro five-membered cyclic deformylase inhibitors of the present invention can effectively inhibit the proliferation of various cancer cells at a concentration of 30 ⁇ M, such as colorectal cancer, lung cancer, and gastric cancer. And liver cancer, etc., highlighting the selective inhibition of cancer cells in the digestive tract (see Tables 4 and 5). Meanwhile, the spirotrienoid and spiro five-membered cyclic peptide deformylase inhibitor of the present invention can also effectively inhibit the proliferation of other various cancer cells such as: Leukemia, cervical cancer, breast cancer, prostate cancer, and osteosarcoma highlight the selective inhibition of leukemia and osteosarcoma cells (see Tables 6 and 7).
- the present invention uses the agar double dilution method recommended by the American College of Clinical and Laboratory Standards Institute (CLSI) antibiotic susceptibility test protocol to determine the minimum inhibitory concentration of the test sample on the tested strain ( Minimal Inhibitory Concentration (MIC), compared with the in vitro antibacterial activity of the control drug LBM415 and levofloxacin, examined 46 synthetic Gram-positive pathogens and Gram-negative bacteria isolated from clinically isolated compounds in the past three years. In vitro antibacterial activity of the pathogen.
- CCSI American College of Clinical and Laboratory Standards Institute
- the present invention uses the CCK-8 method to determine the spirotricyclic and spiro five-membered cyclic deformylase inhibitors described in Examples 1-80 for colorectal cancer cells, lung cancer cells, gastric cancer cells, and liver cancer.
- the proliferation inhibitory effect of four tumor cells such as cells. Firstly, 80 compounds were tested for anti-colorectal cancer cell proliferation activity, and then 30 anti-colorectal cancer cells with good proliferative activity were selected. Four kinds of compounds such as colorectal cancer cells, lung cancer cells, gastric cancer cells and liver cancer cells were examined. The inhibitory activity of tumor cells (see Table 5); the half effect inhibitory concentration (IC 50 ) of 6 of the compounds with better activity was further determined (see Table 6). The compound having the best activity of Example 64 was further tested for its inhibitory activity against other tumor cells including T cell leukemia, acute myeloid leukemia, cervical cancer, breast cancer, prostate cancer and osteosarcoma (see Table 7).
- HCT-116 colorectal cancer cell line A549 lung cancer cell line, MGC-803 gastric cancer cell line, BEL-7402 liver cancer cell line, Jurkat T cell leukemia cell line, HL60 acute myeloid leukemia cell line, Hela cervical cancer Cell line, MCF-7 breast cancer cell line, PC3 prostate cancer cell line and SAOS-2 osteosarcoma cell line were prepared as single cell suspension, 180 ⁇ L inoculated in 96-well culture plate, CO 2 incubator (37 ° C, 5%) CO 2 , 95% air) overnight culture;
- the concentration of the single cell suspension is 2000 cells/well
- HCT-116 cells were treated with McCoy's 5A medium (10% containing newborn bovine serum, 1% double antibody), and A549 and PC3 cells were treated with F12 medium (10% containing newborn bovine serum, 1% double).
- Anti-), MGC-803, BEL-7402, SAOS-2, Jurkat and HL60 cells were cultured in 1640 medium (10% containing newborn bovine serum, 1% double antibody), and MCF-7 and Hela cells were cultured in DMEM high glucose medium ( 10% contains newborn bovine serum, 1% double antibody);
- the calculation method of the inhibition rate in the step (2) is 1-(A drug treatment group- A blank control )/(A drug-free treatment group- A blank control ), and A is absorbance.
- the compounds are respectively formulated into 30 ⁇ M, 10 ⁇ M, 3.3 ⁇ M, 1.1 ⁇ M, 0.37 ⁇ M, 0.12 ⁇ M, 0.04 ⁇ M, and 0.013 ⁇ M, and added to the cells, and after 72 hours, the steps are followed (3).
- the absorbance was measured, and the inhibition rate of tumor cell growth by different concentrations of the compound was calculated.
- the data was analyzed by GraphPad Prism 5, and the IC 50 value (the IC 50 value is the corresponding compound concentration when the inhibition rate reached 50%) was calculated.
- Step 1 After adding DMF to the acid, add 2.1 equivalents of N-methylimidazole under ice bath, then slowly add 1 equivalent of MsCl, stir for 15 min, then add 1 equivalent of Boc-protected amine, and the reaction is detected by TLC. After completion of the reaction, the EA was diluted, washed with 10% citric acid, and the aqueous phase was extracted twice with 50 mL of EA. The organic phase was combined and washed sequentially with saturated sodium hydrogen carbonate and brine, and the organic phase was concentrated.
- Step 2 A solution of HCl/MeOH is added to the product obtained in the above step. After the reaction is completed, the product is obtained.
- R 1 is n-butyl, cyclopentylmethyl
- R 2 is an aromatic or aromatic heterocyclic ring, a heterocyclic ring, an alkyl group.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 1H-pyrazol-3-yl, 5-fluoropyridine 1-oxide-2-yl, 5-(uncle Butyl)isoxazol-3-yl, 6-methyl-N-(4-(pyridin-3-yl)pyrimidin-2-yl)benzene-1-amino-3-yl, 3-fluoropyridine-2 -yl, 5-methylthiazol-2-yl, 3-(pyridin-3-yl)phenyl, N-(pyrimidin-2-yl)benzene-1-amino-3-yl, 4-(pyridine-3 -yl)pyrimidin-2-yl, 4-morpholinephenyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyrimidinyl, 1-methyl-1H-pyrazol-4-yl, 5-iso Oxazolyl, cyclopropyl, 4-methylthiazol
- n 2 or 4
- R 1 is n-butyl, cyclopentylmethyl
- R 2 is an aromatic ring or an aromatic heterocyclic ring.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 2-benzimidazolyl, 1,3,4-oxadioxazole, 1,2,4-oxo Oxazole, 1,3,4-triazole.
- Step 3 First dissolve the acid in DMF, add 5 times equivalent of DIPEA, HATU (1.05 equivalents) in the ice bath, stir for 15 minutes, then add the synthesized amine (1.0 equivalents), the reaction is completed, the reaction After the solution was diluted with ethyl acetate, the mixture was washed twice with 10% aqueous citric acid, and the aqueous phase was extracted twice with ethyl acetate. The organic phase was combined and washed with saturated sodium hydrogen carbonate and saturated sodium chloride. The sodium was dried, filtered and concentrated to give a crude material which was taken directly to the next step.
- Step 4 The crude material was dissolved in dichloromethane (2 mL)EtOAc After the reaction was stirred at room temperature for 2 hours, it was neutralized with a saturated aqueous solution of sodium carbonate, and the organic phase was obtained. The aqueous phase was extracted once with methylene chloride.
- R 1 is n-butyl, cyclopentylmethyl
- R 2 is an aromatic or aromatic heterocyclic ring, a heterocyclic ring, an alkyl group.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 1H-pyrazol-3-yl, 5-fluoropyridine 1-oxide-2-yl, 5-(uncle Butyl)isoxazol-3-yl, 6-methyl-N-(4-(pyridin-3-yl)pyrimidin-2-yl)benzene-1-amino-3-yl, 3-fluoropyridine-2 -yl, 5-methylthiazol-2-yl, 3-(pyridin-3-yl)phenyl, N-(pyrimidin-2-yl)benzene-1-amino-3-yl, 4-(pyridine-3 -yl)pyrimidin-2-yl, 4-morpholinephenyl, 2-pyrazinyl, 3-pyridazinyl, 4-pyrimidinyl, 1-methyl-1H-pyrazol-4-yl, 5-iso Oxazolyl, cyclopropyl, 4-methylthiazol
- n 2 or 4
- R 1 is n-butyl, cyclopentylmethyl
- R 2 is an aromatic ring or an aromatic heterocyclic ring.
- n 2 and 4;
- R 1 is n-butyl, cyclopentylmethyl;
- R 2 is 2-benzimidazolyl, 1,3,4-oxadioxazole, 1,2,4-oxo Oxazole, 1,3,4-triazole.
- Step 3 As in step 3 of the hydroxamic acid series.
- Step 4 As in step 4 of the hydroxamic acid series.
- Step 5 The acid obtained in the previous step was dissolved in DMF, and 5 times equivalent of DIPEA, HATU (1.05 equivalents) was added to the ice bath, and after stirring for 15 minutes, 3 times of hydroxylamine hydrochloride was added. The reaction was completed and preparative HPLC purification was carried out. The final product is obtained.
- Embodiments 6, 22, 25, 29, 32, 33, 34, 36, 38, 40, 46, 51, 55, 57, 61, 62, 63, 64, 65, 66, 67, 70, 71, 72 were selected. 30, 73, 75, 76, 77, 78, 79, 30 compounds that have a good inhibitory effect on the proliferation of colorectal cancer cells HCT116 (inhibition rate ⁇ 30%), and examined them for other cancer cells such as lung cancer cells, gastric cancer cells, Proliferative inhibition rate activity of liver cancer cells. The results showed that 13 compounds in Examples 46, 57, 62, 63, 64, 65, 66, 70, 72, 73, 76, 78, 79 were at 30 ⁇ M.
- the inhibition rate of proliferation of lung cancer cells A549 was more than 50%, and the compound of Example 76 had the highest proliferation inhibition rate of 94%;
- Examples 6, 29, 38, 40, 62, 63, 64, 65, 66 The inhibition rate of proliferation of gastric cancer cells MGC-803 by the 14 compounds of 72, 73, 75, 76, 79 was more than 50% at a concentration of 30 ⁇ M, and the compound of Example 29 had the highest inhibition rate of 78%;
- Examples 6, 22, 25, 29, 33, 34, 36, 38, 40, 46, 51, 55, 57, 62, 63, 64, 65, 66, 67, 70, 72, 73, 75, 76, 77 The inhibition rate of 27 compounds of 78, 79 to the liver cancer cell BEL-7402 at a concentration of 30 ⁇ M exceeded 50%, and the compounds of Examples 64, 65 and 73 had the highest proliferation inhibition rate of 81%.
- Step 2 The operation is as follows: Step 1 in the synthesis of the general formula (X1). After adding 20 mL of DMF to the acid (2.05 g, 8.5 mmol), N-methylimidazole (1.54 g, 18.7 mmol) was added under ice-cooling, then MsCl (1.07 g, 9.4 mmol) was slowly added dropwise, and the mixture was stirred for 15 min.
- Step 3 20 mL and 10 mL of a 5 M HCl/MeOH solution were added to the oil, and the mixture was reacted at room temperature overnight. The reaction was completed and dried to give an oil (2.0 g, yield of 100% in two steps).
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows, Step 4 in the synthesis of the general formula (X2). The yield in two steps was 18%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation was as follows: Step 2 (2.1 g, white solid, yield of 68% in two steps) of the formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The obtained oil (242 mg, 0.46 mmol) was dissolved in ethyl acetate (3mL), EtOAc (EtOAc, EtOAc, EtOAc Mixture Stir at room temperature for 2 hours. After completion of the reaction, the reaction was quenched with sodium thio sulfate and ethyl acetate. The organic phase was dried and concentrated to give a crude material.
- Step 5 The operation is as follows, Step 4 in the synthesis of the general formula (X2). The three-step yield is 45%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The procedure was as follows: Step 2 (1.4 g, white solid, yield 47%) in the formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation was as follows: Step 2 (3.1 g, yellow solid, yield of 66% in two steps) of formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation was carried out as in Step 2 (1.7 g, white solid, two-step yield 45%) of formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The procedure is as follows: Step 2 (680 mg, white solid, 100% yield in two steps) of the formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 Add ethylene glycol to the mixture of 3-bromopyridine (2.37 g, 15 mmol), 3-aminophenylboronic acid monohydrate (2.32 g, 15 mmol) and Pd(PPh 3 ) 4 (520 mg, 0.45 mmol). Ether (30 mL) and water (15 mL). After completion of the reaction, the solvent was evaporated, and the residue was evaporated,jjjjjjjjjjjjjjjjjj
- Step 2 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- (S)-5-(tert-Butyl)-5-azaspiro[2.4]heptane-6-carboxylic acid 950 mg, 3.94 mmol
- DMF 5 mL
- NMI 0.7 mL, 8.67
- MsCl 451 mg, 3.94 mmol
- 3-(pyridin-3-yl)aniline (670 mg, 3.94 mmol) was added.
- TLC monitors the progress of the reaction. After completion of the reaction, the reaction mixture was diluted with EtOAc EtOAc EtOAc. The organic phases were combined and washed with a saturated aqueous solution of sodium carbonate and then brine.
- the crude product was obtained by concentrating under reduced pressure under vacuo.
- Step 3 The crude was dissolved in 10 mL of ethyl ether. EtOAc (EtOAc) The precipitate was collected and dried to give a brown solid (yield: EtOAc, EtOAc,
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 Xantphos (347 mg, 3 mol%), Pd 2 (dba) 3 (366 mg, 2 mol%) 2-aminopyrimidine (1.92 g, 20 mmol) and sodium tert-butoxide (1.92 g, 20 mmol) were added to the reaction flask. The system was replaced three times with nitrogen. Toluene solution (20 mL) of 3-bromo-1-nitrobenzene (4.04 g, 20 mmol) was added to the reaction. Heat to 95 ° C and react for 18 hours.
- Step 2 Trifluoroethanol (20 mL) containing 10% Pd/C (50 mg), N-(3-nitrophenyl)pyrimidin-2-amine (1.3 g, 6.0 mmol) EtOAc (1 mL). The solution was heated to 80 ° C for 5 hours. After completion of the reaction, the catalyst was filtered off, and the obtained filtrate was concentrated to give the product (1.1 g, 100% yield).
- Step 3 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- (S)-5-(tert-Butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (965 mg, 4.0 mmol) was dissolved in DMF (8 mL).
- MsCl 460 mg, 4.0 mmol
- N-(2-pyrimidinyl)benzene-1,3-diamine 745 mg, 4.0 mmol
- the mixture was diluted with EtOAc EtOAc.
- the combined organic phases were washed with saturated sodium carbonate and brine. Concentration under reduced pressure gave a yellow solid.
- Step 4 The solid was dissolved in a 2:1 DCM/TFA (8 mL / 4 mL) solution and allowed to react at room temperature for 1 hour. After the reaction was completed, a few drops of water were added and neutralized with solid sodium carbonate solid. After drying over anhydrous Na 2 SO 4 , the insoluble solid was filtered, and then the filtrate was concentrated to give a free base (500 mg, 50% yield in two steps).
- Step 5 The operation is as follows, Step 3 in the synthesis of the general formula (X2).
- Step 1 Suspension of 3-dimethylamino-1-(3-pyridyl)-2-propenyl-1-one (1.76 g, 10.0 mmol) in n-butanol (20 mL) followed by hydrazine hydrochloride (1.15 g) , 12.0 mmol) and NaOH (480 mg, 12.0 mmol). The mixture was heated to 120 ° C and allowed to react overnight. The precipitated solid was collected, washed with water (50 mL) and evaporated.
- Step 2 Dissolve (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (2.0 g, 8.5 mmol) in DMF (20 mL). g, 25.5 mmol), cooled to 0 ° C, EtOAc (EtOAc, EtOAc) Then 4-(3-pyridyl)pyrimidin-2-amine (980 mg, 5.7 mmol) and lithium chloride (721 mg, 17.0 mmol) were added. After stirring at room temperature for 48 hours, the reaction mixture was diluted with EtOAc EtOAc. The organic phase was washed with saturated sodium carbonate and brine. After concentrating under reduced pressure, EtOAc m.
- Step 3 The product obtained was separated and dissolved in 10 mL of diethyl ether. 5 mL HCl/MeOH (5M). The precipitate was collected and dried to give a white solid ( ⁇ /RTI> ⁇ RTIgt;
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 Place (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (2.4 g, 10 mmol) in a flask and add N,N-dimethyl
- the formamide DMF 25 mL
- N-methylimidazole NMI 1.8 mL, 22 mmol
- methanesulfonyl chloride 0.78 mL, 10 mmol
- o-phenylenediamine 2.8 g, 20 mmol
- reaction solution was diluted with ethyl acetate, and the organic phase was washed three times with 10% aqueous citric acid to remove excess o-phenylenediamine. Finally, the organic phase was dried and concentrated to give a crude pink product as a foamy solid.
- Step 2 The obtained solid was dissolved in 20 mL of methyl tert-butyl ether, and 4 mL of acetic acid was added. The solution was heated to reflux for 3 hours, and after completion of the reaction, it was neutralized with a saturated Na 2 CO 3 solution to neutral. The organic layer was isolated and concentrated to give the product.
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 3-cyanopyridine (5.2g, 50mmol) was dissolved in 50mL of ethanol, followed by addition of hydroxylamine hydrochloride (4.2g, 60mmol), NaHCO 3 (5.0g, 60mmol) and water (3mL). The mixture was heated to reflux for 12 hours. Anhydrous Na 2 SO 4 was added , filtered, and diluted with diethyl ether (50 mL) to give white crystals (6.6 g, 96% yield).
- Step 2 Dissolve (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (2.49 g, 10.3 mmol) in DMF (25 mL) then N , N'-carbonyldiimidazole (CDI, 1.75 g, 10.3 mmol). After the mixture was stirred at room temperature for 8 hours, N- hydroxy-(3-pyridyl)carbazide (1.41 g, 10.3 mmol) was added, and then another equivalent of CDI (1.75 g, 10.3 mmol) was added. The reaction was heated to 90 ° C under a nitrogen atmosphere for 12 hours. After cooling, it was diluted with EtOAc (EtOAc)EtOAc. The collected organic phase was washed with aq. EtOAc (EtOAc) EtOAc.
- EtOAc EtOAc
- Step 3 An oil (2.1 g, 5.67 mmol) was dissolved in 10 mL diethyl ether. A small amount of water was added to the reaction mixture, and the acid was neutralized by adding sodium carbonate solid. After the bubbles disappeared, the solid was filtered off, and the filtrate was concentrated to give a free base (1.2 g, 80% yield).
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 (S)-5-(tert-Butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (1.0 g, 4.14 mmol) was dissolved in DMF (10 mL). M, Cl (475 mg, 4.14 mmol) was added dropwise at 0 ° C and stirred for 15 min. Then 2-pyridineformylhydrazide (569 mg, 4.14 mmo) was added. After 6 hours of reaction, it was diluted with EtOAc EtOAc (EtOAc) The combined organic phases were washed with EtOAc EtOAc (EtOAc)
- Step 3 An oil (1.4 g, 4.13 mmol) was dissolved in 2:1 DCM/TFA (6mL / 3mL), and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, a few drops of water were added to the reaction solution, followed by neutralization with a sodium carbonate solid. It was dried over anhydrous sodium sulfate, and the insoluble material was filtered. The filtrate was concentrated to dryness to dryness (1.0 g, 100% yield).
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 Dissolve (S)-5-(tert-butoxycarbonyl)-5-azaspiro[2.4]heptane-6-carboxylic acid (2.4 g, 10 mmol) in DMF (25 mL). , 22 mmol). After cooling to 0 ° C, MsCl (0.78 mL, 10 mmol) was added dropwise and stirred for 15 min. 2-Pyridinium (1.1 g, 10 mmol) was then added to the solution. The reaction was quenched with EtOAc (EtOAc)EtOAc. The organic phase was washed with saturated brine. The crude product was obtained by EtOAc EtOAc (EtOAc)
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 22%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation was as in Step 1 of Example 1.
- Step 2 The operation is as in step 2 of the embodiment 1.
- Step 3 The operation is as in Step 3 of Example 1.
- Step 4 The product in step 3 is dissolved in THF, then 2.5 times equivalent of NaH is added. After 30 minutes, 1.5 times equivalent of dimethylaminochloroethane is added, and the reaction is refluxed at 50 ° C. After completion of the reaction, water, EA, and extraction are added. After drying over anhydrous sodium sulfate, the column was concentrated to give the product.
- Step 5 The operation is as follows, Step 3 in the synthesis of the general formula (X2).
- Step 6 Operation As in Step 4 of the synthesis of the formula (X2), a white solid obtained by lyophilization was obtained in a two-step yield of 27%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 Operation as in the synthesis of step 1 in the general formula (X1)
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation was as in Step 1 of Example 1.
- Step 2 The operation is as in step 2 of the embodiment 1.
- Step 3 The operation is as in Step 3 of Example 1.
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 36%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 25%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 22%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 25%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 29%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 23%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 23%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 29%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 32%.
- Step 1 The operation was as in Step 1 of Example 1.
- Step 2 The operation is as in step 2 of the embodiment 1.
- Step 3 The operation is as in Step 3 of Example 1.
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 6 The operation is as follows: Step 5 in the synthesis of the general formula (X2), a three-step yield of 17%.
- Step 1 The operation was as in Step 1 of Example 1.
- Step 2 The operation is as in step 2 of the embodiment 1.
- Step 3 The operation is as in Step 3 of Example 1.
- Step 1 The operation was as in Step 1 of Example 1.
- Step 2 The operation is as in step 2 of the embodiment 1.
- Step 3 The operation is as in Step 3 of Example 1.
- Step 4 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 6 The operation is as follows: Step 5 in the synthesis of the general formula (X2), a three-step yield of 17%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 23%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows, Step 5 in the synthesis of the general formula (X2), a three-step yield of 27%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 24%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 18%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 Operation As in the synthesis of Step 5 in the formula (X2), the yield in three steps was 19%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
- Step 2 The operation is as follows: Step 2 in the synthesis of the general formula (X1).
- Step 3 The operation is as follows: Step 3 in the synthesis of the general formula (X2).
- Step 4 The operation is as follows, Step 4 in the synthesis of the general formula (X2).
- Step 5 The operation is as follows: Step 5 in the synthesis of the general formula (X2), the yield in three steps is 20%.
- Step 1 The operation is as follows: Step 1 in the synthesis of the general formula (X1).
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Abstract
本发明公开了一类新型的含螺三元环、螺五元环类肽脱甲酰基酶抑制剂的抗菌活性和抗肿瘤活性。本发明的含螺三元环、螺五元环类螺环类肽脱甲酰基酶抑制剂可作为一类新型的抗菌剂，通过抑制细菌蛋白质合成中所需肽脱甲酰基酶的活性对多种抗生素耐药的革兰氏阳性菌菌株都有效，且不影响人体的主要蛋白质合成过程，从而选择性杀死细菌；本发明的含螺三元环、螺五元环类螺环类肽脱甲酰基酶抑制剂还可作为一类新型的抗癌药物，通过抑制癌症细胞线粒体中的肽脱甲酰基酶，可以影响细胞的能量平衡：从而使得线粒体膜去极化，ATP耗竭和促使细胞凋亡，在较低浓度下对多种癌细胞菌株如结直肠癌、白血病、肺癌、胃癌、***、乳腺癌、***癌、肝癌和骨肉瘤等具有较好的抑制活性。
Description
本发明属于抗菌和抗癌药物技术研究领域，涉及一类肽脱甲酰基酶抑制剂，具体涉及新型螺三元环、螺五元环类肽脱甲酰基酶抑制剂。
抗生素(antibiotics)是指一系列能在一定浓度下对病原体有抑制和杀灭作用的化学物质，其包括由微生物、动植物产生的代谢物，也包括用化学方法合成或半合成的化合物。抗生素不仅指抗菌物质，抗肿瘤、抗病毒、抗寄生虫等物质亦属抗生素范畴。抗生素是使我们能够延长寿命、更健康地生活和受益于现代医学的重要支柱。
随着抗生素的大量出现和广泛使用，抗生素耐药性问题日益凸显，产生耐药性的速度越来越快，时间之快，耐药谱之广，令人乍舌。
全球超过95％金黄色葡萄球菌几乎对所有的盘尼西林都耐药，对90％的甲氧西林耐药；超过80％的大肠杆菌已对三分之一的第三代头孢菌素不敏感，对90％的氟喹諾酮耐药。不仅如此，很多细菌开始产生了多重耐药。在临床中，甲氧西林耐药的金黄色葡萄球菌对阿莫西林、左氧氟沙星、利福平、甚至是万古霉素都产生了耐药；大肠杆菌和克雷伯氏肺炎菌形成了新的耐药机制，产生新德里金属-β-内酰胺酶1(NewDelhi metallo-β-lactamase 1,NDM-1)，几乎对所有的β-内酰胺类药物都耐药。
化疗是目前临床上治疗恶性肿瘤的最重要手段之一，然而由于肿瘤细胞常常会对化疗药物产生耐药而导致患者对治疗不再敏感，最终导致化疗失败。尽管表皮生长因子受体(EGFR)抑制剂易瑞沙和特罗凯等在治疗非小细胞肺癌中取得了很大的成功，但其耐药问题日益突出，耐药一半源自EGFR T790M突变。肝细胞癌是世界上最常见的恶性癌症之一，对化疗药物也通常表现出高度耐受性。近一半的乳腺癌从一开始治疗就对赫赛汀产生了耐药。
鉴于抗生素耐药情况日益严峻，开发新的作用机制的抗生素迫在眉睫，而肽脱甲酰基酶抑制剂则是近年来研究比较热的新作用靶点之一。肽脱甲酰基酶(peptide deformylase,PDF)是一种金属蛋白酶，不仅广泛存在于细菌体内，也存在于恶性疟原虫和人体中。在细菌合成蛋白的过程中，PDF可以脱除甲硫氨酸上的甲酰基，从而使细菌能够合成出功能化的蛋白质。脱除甲酰基的过程是细菌合成蛋白的必须过
程。而人体细胞主要蛋白合成过程却不依赖甲酰基脱除的过程，细菌与人体合成蛋白过程的这种差异使细菌PDF有可能成为抗菌药物作用的新靶点。PDF抑制剂通过与PDF酶的螯合，使细菌在合成蛋白过程中脱甲酰基步骤受阻，从而起到选择性抑制细菌生长的目的。与正常人体细胞比，多种癌症细胞如：结直肠癌、肺癌、***癌等细胞的人PDF基因是过度表达的，通过抑制癌细胞线粒体中的肽脱甲酰基酶，可以影响细胞的能量平衡：从而使得癌细胞线粒体膜去极化，ATP耗竭和促使细胞凋亡。PDF抑制剂可发展成为具有较好抗菌活性的广谱抗菌剂和抗癌活性的抗癌药物。
PDF抑制剂所包含的结构：金属鳌合基团(Metal Binding Group,MBG)和含P1、P2、P3结构部分的拟肽或非肽类骨架(式(a)PDF抑制剂一般结构)。
尽管有诸多PDF抑制剂被开发出来，用于临床前的研究，甚至有些化合物进入了临床(如式(b)～(d))，但是由于化合物本身性质的原因导致活性不佳，或表现出临床毒性，最终都未能成功上市。比如：放线菌素Actinonin是第一个被发现的PDF抑制剂，其表现了良好的抗革兰氏阳性菌和阴性菌的活性，但是由于体内代谢的不稳定，最终导致无体内的活性。LBM415，进入抗菌活性的一期临床(Phase I)试验，具广谱活性，但是发现高剂量时会引起高铁血红蛋白症(Clin.Pharmacol.Ther.2011,90,256)；GSK1322322，也被FDA终止抗菌活性的一期临床的研究，原因类似，出现代谢活性化合物，造成对机体的毒性(临床研究终止原因见ClinicalTrials.gov中的项目编号NCT01818011)。
发明内容
本发明提供了一类结构新颖、活性较佳、毒性小的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂，用于抑制对现有抗生素耐药的细菌，包括革兰氏阳性菌，如金黄色葡萄球菌、表皮葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌，和严重影响人类健康的革兰氏阴性菌，如卡他莫拉菌等；本发明还报道了所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂选择性抑制癌细胞如结直肠癌、肺癌、胃癌和肝癌等增殖的作用。
本发明的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂，属于抗生素，含以下式(1)～(4)四类结构：
第一类：
式(1)中，n＝2-4；R1为正丁基，环戊甲基；R2为芳香环，芳香杂环，杂环，烷基；R3为氢或烷基。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为1H-吡唑-3-基、5-氟吡啶1-氧化物-2-基、5-(叔丁基)异恶唑-3-基、6-甲基-N-(4-(吡啶-3-基)嘧啶-2-基)苯-1-氨基-3-基、3-氟吡啶-2-基、5-甲基噻唑-2-基、3-(吡啶-3-基)苯基、N-(嘧啶-2-基)苯-1-氨基-3-基、4-(吡啶-3-基)嘧啶-2-基、4-***啉苯基、2-吡嗪基、3-哒嗪基、4-嘧啶基、1-甲基-1H-吡唑-4-基、5-异恶唑基、环丙基、4-甲基噻唑-2-基、2-恶唑基、5-甲基异恶唑-3-基、2-噻唑基、1,3,4-硫二唑-2-基、5-(三氟甲基)噻唑-2-基、双乙基砜基、苯并噻唑-2-基、3-甲酸甲酯-2-噻吩基；R3为氢。
进一步优选地，所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂为实施例1-9、14-23、25-41中的化合物。
第二类：
式(2)中，n＝2-4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为芳香杂环。
进一步优选地，n＝2和4；R1为正丁基，环戊甲基；R2为2-苯并咪唑基、1,3,4-氧二恶唑、1,2,4-氧二恶唑、1,3,4-三氮唑。
进一步优选地，所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂为实施例10-13、24、42中的化合物。
第三类：
式(3)中，n＝2-4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环，杂环，烷基；R3为氢或烷基。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为芳香杂环；R3为氢。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为1H-吡唑-3-基、5-氟吡啶1-氧化物-2-基、5-(叔丁基)异恶唑-3-基、6-甲基-N-(4-(吡啶-3-基)嘧啶-2-基)苯-1-氨基-3-基、3-氟吡啶-2-基、5-甲基噻唑-2-基、3-(吡啶-3-基)苯基、N-(嘧啶-2-基)苯-1-氨基-3-基、4-(吡啶-3-基)嘧啶-2-基、4-***啉苯基、2-吡嗪基、3-哒嗪基、4-嘧啶基、1-甲基-1H-吡唑-4-基、5-异恶唑基、环丙基、4-甲基噻唑-2-基、2-恶唑基、5-甲基异恶唑-3-基、2-噻唑基、1,3,4-硫二唑-2-基、5-(三氟甲基)噻唑-2-基、双乙基砜基、苯并噻唑-2-基、3-甲酸甲酯-2-噻吩基；R3为氢。
进一步优选地，所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂为43-57、61-74、77-80中的化合物。
第四类：
式(4)中，n＝2-4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为芳香杂环。
进一步优选地，n＝2和4；R1为正丁基，环戊甲基；R2为2-苯并咪唑基、1,3,4-氧二恶唑、1,2,4-氧二恶唑、1,3,4-三氮唑。
进一步优选地，所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂为58-60、75、76中的化合物。
本发明的式(1)～(4)螺三元环类、螺五元环类肽脱甲酰基酶抑制剂的分子量介于300-600；可溶于二氯甲烷、丙酮、乙腈、甲醇、乙醇、N,N‐二甲基甲酰胺、二甲基亚砜等溶剂；微溶于***、水等，不溶于石油醚；通常为无色或微黄色粉末或泡沫。
本发明还提出了将所述式(1)～(4)所示螺三元环类、螺五元环类肽脱甲酰基酶抑制剂在抑制细菌肽脱甲酰基酶中的应用。
其中，所述细菌包括金黄色葡萄球菌，表皮葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌、卡他莫拉菌。
本发明采用左氧氟沙星和LBM415作为对照化合物，对所合成的80个新化合物进行了四十六株临床分离菌株(菌株见表1)的体外抗菌活性筛选(80个化合物一览表见表2)。
本发明还提出了将所述式(1)～(4)所示螺三元环类、螺五元环类肽脱甲酰基酶抑制剂在制备抗肿瘤药物中的应用。
其中，所述肿瘤是指结直肠癌、白血病、肺癌、胃癌、***、乳腺癌、***癌、肝癌和骨肉瘤等。
本发明采用cck-8法测定了化合物对结直肠癌、白血病、肺癌、胃癌、***、乳腺癌、***癌、肝癌和骨肉瘤等九种肿瘤细胞的增殖抑制效果。
本发明还提出了所述式(1)～(4)所示螺三元环类、螺五元环类肽脱甲酰基酶抑制剂在制备抗寄生虫的药物中的应用。
其中，所述抗寄生虫是指所述螺三元环、螺五元环类肽脱甲酰基酶抑制剂通过抑制寄生虫肽脱甲酰基酶来抑制寄生虫的生长。
本发明还提出了一种药物组合物，所述药物组合物包括如上所述的螺三元环、螺五元环类肽脱甲酰基酶抑制剂以及药学上可接受的载体。
其中，所述药学上可接受的载体是指药学领域常规的药物载体，例如：稀释剂、赋形剂和水等，填充剂如淀粉、蔗糖，乳糖、微晶纤维素等；粘合剂如纤维素衍生物、藻酸盐、明胶和聚乙烯吡咯烷酮；润湿剂如甘油；崩解剂如羧甲基淀粉钠，羟丙纤维素，交联羧甲基纤维素，琼脂、碳酸钙和碳酸氢钠；吸收促进剂如季铵化合物；表面活性剂如十六烷醇，十二烷基硫酸钠；吸附载体如高龄土和皂粘土；润滑剂如滑石粉、硬脂酸钙和镁、微粉硅胶和聚乙二醇等。另外还可以加入如香味剂、甜味剂等。
本发明中，所述抑制剂或药物组合物的给药方式包括口服、注射、植入的方式等。
本发明中，所述抑制剂或药物组合物可以以药片、液体或胶囊等的形式给药。
本发明新型的含螺三元环、螺五元环类肽脱甲酰基酶抑制剂具有抗菌活性和抗肿瘤活性。本发明的螺环类肽脱甲酰基酶抑制剂的作用机制是：通过抑制细菌蛋白质合成中所需肽脱甲酰基酶的活性，且不影响人体的主要蛋白质合成过程，从而选择性杀死细菌；通过抑制癌症细胞线粒体中的人肽脱甲酰基酶，可以影响细胞的能量平衡：从而使得线粒体膜去极化，ATP耗竭和促使细胞凋亡。本发明的抑制剂由于新颖的作用机制，使得其不易产生如市售抗生素的耐药性。该类肽脱甲酰基酶抑制剂不仅可作为一类新型的抗菌剂，对多种抗生素耐药的革兰氏阳性菌菌株都有效，如金黄色葡萄球菌，表皮葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌；并对革兰氏阴性菌如卡他莫拉菌等有效。还可作为一类新型的抗癌药物，对多种癌细胞菌株如结直肠癌、肺癌、胃癌和肝癌具有低微摩尔的抑制活性。
与现有技术化合物相比，本发明具有明显的技术优势：
(1)本发明的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂通过分子对接研究
和实验抗菌活性筛选等方法发现的(80个化合物抗菌活性见表3所示)。通过分子对接研究，发现了用于活性提高的配体与靶蛋白间的额外的疏水作用力，即抑制剂的螺三元环可以和蛋白上的精氨酸残基形成较强的作用力，这一结论得到实验的支持，如实施例2中的化合物对耐甲氧西林葡萄球菌(MRSA)的最低抑菌浓度MIC达到0.125-0.25μg/mL，而LBM415的MIC＝0.5-1μg/mL，本发明实施例2中的化合物提高了四倍，有了显著的提高(见对照实验一中表8)；通过实验发现，螺五元环的引入对螺五元环类抑制剂活性的提高也起到了显著影响，如实施例77、实施例78、实施例79等等；
(2)本发明采用芳香酰胺修饰的策略，使得改造后一些化合物(如实施例3、实施例4、实施例5、实施例6、实施例7、实施例8、实施例15、实施例16、实施例19、实施例21、实施例22、实施例23、实施例25、实施例26、实施例27、实施例28、实施例29、实施例30、实施例33、实施例38等等)对受试菌株的活性有了实质性的提高，如对耐甲氧西林葡萄球菌(MRSA)的最低抑菌浓度MIC达到<0.008-0.06μg/mL，远远优于先导化合物LBM415(MIC＝0.5-1μg/mL)和已上市抗生素药物左氧沙星(MIC＝16>128μg/mL)；以唑类化合物为酰胺生物电子等排体的策略的应用，不仅使化合物活性得到了保持，大部分化合物MIC＝0.5-2μg/mL，如：实施例10、实施例11、实施例12、实施例13、实施例75等；关键是本发明增加了化合物的稳定性，也降低了化合物的代谢毒性(见对照实验二表9)；
(3)本发明的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂体外抗菌实验表明，其不仅对革兰氏阳性菌如金黄色葡萄球菌、表皮葡萄球菌、耐药金黄色葡萄球菌、耐药的表皮葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌有效，最低抑制浓度仅有<0.008μg/mL，其活性比先导化合物LBM415高60倍，如实施例3、实施例4、实施例6、实施例33、实施例38、实施例64、实施例72、实施例73等；而且对严重影响人类健康的革兰氏阴性菌卡他莫拉菌等表现出很好的活性，最低抑菌浓度低至0.06μg/mL，如实施例22、实施例33等(见对照实验三表10)。
(4)本发明的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂在30μM的浓度条件下可以很有效地抑制多种癌细胞的增殖如：结直肠癌、肺癌、胃癌和肝癌等，突出表现出对消化道癌细胞的选择性抑制(见表4和5)。同时，本发明的螺三元环类、螺五元环类肽脱甲酰基酶抑制剂也可以很有效地抑制其他多种癌细胞的增殖如：
白血病、***、乳腺癌、***癌和骨肉瘤等，突出表现出对白血病、骨肉瘤细胞的选择性抑制(见表6和7)。
具体实施方法
结合以下具体实施例，对本发明作进一步的详细说明。实施本发明的过程、条件、实验方法等，除以下专门提及的内容之外，均为本领域的普遍知识和公知常识，本发明没有特别限制内容。
实验方法
1.本发明采用美国临床和实验室标准协会(Clinical and Laboratory Standards Institute,CLSI)抗菌药物敏感性试验操作规程推荐的琼脂二倍稀释法测定受试样品对所试菌株的最低抑菌浓度(Minimal Inhibitory Concentration，MIC)，通过与对照药LBM415、左氧沙星的体外抗菌活性比较，考察了全部合成化合物对近三年临床分离的四十六株革兰阳性致病菌和革兰阴性致病菌的体外抗菌活性。
表1.体外抗菌活性筛选的受试临床分离菌株
2.本发明采用CCK-8法测定了实施例1-80中所述螺三元环类、螺五元环类肽脱甲酰基酶抑制剂对结直肠癌细胞、肺癌细胞、胃癌细胞、肝癌细胞等四种肿瘤细胞的增殖抑制效果。首先测定了80个化合物抗结直肠癌细胞增殖的活性，然后优选30个抗结直肠癌细胞增殖活性好的化合物，考察了其对结直肠癌细胞、肺癌细胞、
胃癌细胞、肝癌细胞等四种肿瘤细胞的抑制活性(见表5)；进一步测定了其中6个活性较好的化合物的半效应抑制浓度(IC50)(见表6)。对活性最好的实施例64制备的化合物，进一步检测了其对包括T细胞白血病、急性髓系白血病、***、乳腺癌、***癌及骨肉瘤等在内的其他肿瘤细胞的抑制活性(见表7)。
(1)将HCT-116结直肠癌细胞株、A549肺癌细胞株、MGC-803胃癌细胞株、BEL-7402肝癌细胞株、Jurkat T细胞白血病细胞株、HL60急性髓系白血病细胞株、Hela***细胞株、MCF-7乳腺癌细胞株、PC3***癌细胞株和SAOS-2骨肉瘤细胞株制成单细胞悬液，180μL接种于96孔培养板中，CO2培养箱(37℃,5％CO2，95％空气)过夜培养；
(2)各螺三元环类、螺五元环类肽脱甲酰基酶抑制剂用DMSO溶解，用相应的细胞培养基配制成终浓度为所需的不同浓度的药物溶液，分别加入到上述细胞(20μL/孔)中，对照组加入1‰DMSO，CO2培养箱中培养72小时；
(3)培养72h后倾去培养液，加入100μL 1:10稀释的CCK-8溶液，37℃孵育2小时后，使用酶标仪SpectraMax M5测450nm处吸光度A，参考波长620nm，计算对肿瘤细胞生长的抑制率。
其中，步骤(1)中，所述单细胞悬液的浓度为2000细胞/孔；
其中，步骤(2)中，HCT-116细胞采用McCoy’s 5A培养基(10％含新生牛血清、1％双抗)，A549和PC3细胞采用F12培养基(10％含新生牛血清、1％双抗)，MGC-803、BEL-7402、SAOS-2、Jurkat和HL60细胞采用1640培养基(10％含新生牛血清、1％双抗)，MCF-7和Hela细胞采用DMEM高糖培养基(10％含新生牛血清、1％双抗)；
步骤(2)中所述抑制率的计算方法为1-(A药物处理组-A空白对照)/(A无药物处理组-A空白对照)，A为吸光度。
测定IC50值时，步骤(2)中，化合物分别配成30μM、10μM、3.3μM、1.1μM、0.37μM、0.12μM、0.04μM、0.013μM，加入到细胞中，处理72h后按步骤(3)测定吸光度，计算不同浓度的化合物对肿瘤细胞生长的抑制率，用GraphPad Prism5分析数据，计算IC50值(IC50值为抑制率达到50％时对应的化合物浓度)。
化合物的制备方法
氨基化合物的合成通式(X1)
步骤1：在酸中加入DMF溶解后，冰浴下加入2.1个当量N-甲基咪唑，然后缓慢滴加1个当量MsCl，搅拌15min后加入1个当量Boc保护的胺，反应用TLC检测，反应完毕，EA稀释，用10％的柠檬酸洗涤，水相用50mLEA萃取2次，合并有机相后依次用饱和碳酸氢钠、饱和食盐水洗涤，有机相浓缩得产物。
步骤2：向上步所得产物中加入HCl/MeOH溶液，待反应完毕后，得产物。
最终化合物的合成通式(X2)
异羟肟酸系列：
式中n＝2或4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环，杂环，烷基。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为1H-吡唑-3-基、5-氟吡啶1-氧化物-2-基、5-(叔丁基)异恶唑-3-基、6-甲基-N-(4-(吡啶-3-基)嘧啶-2-基)苯-1-氨基-3-基、3-氟吡啶-2-基、5-甲基噻唑-2-基、3-(吡啶-3-基)苯基、N-(嘧啶-2-基)苯-1-氨基-3-基、4-(吡啶-3-基)嘧啶-2-基、4-***啉苯基、2-吡嗪基、3-哒嗪基、4-嘧啶基、1-甲基-1H-吡唑-4-基、5-异恶唑基、环丙基、4-甲基噻唑-2-基、2-恶唑基、5-甲基异恶唑-3-基、2-噻唑基、1,3,4-硫二唑-2-基、5-(三氟甲基)噻唑-2-基、双乙基砜基、苯并噻唑-2-基、3-甲酸甲酯-2-噻吩基。
式中n＝2或4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为2-苯并咪唑基、1,3,4-氧二恶唑、1,2,4-氧二恶唑、1,3,4-三氮唑。
步骤3:首先将酸溶解在DMF中，冰浴下依次加入5倍当量的DIPEA，缩合剂HATU(1.05个当量)，搅拌15分钟后加入所合成的胺(1.0个当量)，反应完毕，反应
液用乙酸乙酯稀释后，加10％的柠檬酸水溶液洗涤2次，水相用乙酸乙酯萃取2次，合并有机相，用饱和碳酸氢钠溶液，饱和氯化钠洗涤，经无水硫酸钠干燥，过滤，浓缩后得粗品，直接用于下一步。
步骤4:将粗品溶于二氯甲烷(2mL)，滴加三氟醋酸(1mL)。在室温搅拌反应2小时后，加饱和碳酸钠水溶液中和，分得有机相。水相用二氯甲烷萃取一次，合并有机相，无水硫酸钠干燥、旋干得粗品。
羟肟酸系列：
式中n＝2或4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环，杂环，烷基。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为1H-吡唑-3-基、5-氟吡啶1-氧化物-2-基、5-(叔丁基)异恶唑-3-基、6-甲基-N-(4-(吡啶-3-基)嘧啶-2-基)苯-1-氨基-3-基、3-氟吡啶-2-基、5-甲基噻唑-2-基、3-(吡啶-3-基)苯基、N-(嘧啶-2-基)苯-1-氨基-3-基、4-(吡啶-3-基)嘧啶-2-基、4-***啉苯基、2-吡嗪基、3-哒嗪基、4-嘧啶基、1-甲基-1H-吡唑-4-基、5-异恶唑基、环丙基、4-甲基噻唑-2-基、2-恶唑基、5-甲基异恶唑-3-基、2-噻唑基、1,3,4-硫二唑-2-基、5-(三氟甲基)噻唑-2-基、双乙基砜基、苯并噻唑-2-基、3-甲酸甲酯-2-噻吩基。
式中n＝2或4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。
优选地，n＝2和4；R1为正丁基，环戊甲基；R2为2-苯并咪唑基、1,3,4-氧二恶唑、1,2,4-氧二恶唑、1,3,4-三氮唑。
步骤3:如同异羟肟酸系列的操作步骤3。
步骤4:如同异羟肟酸系列的操作步骤4。
步骤5:取上一步所得酸溶于DMF中，冰浴下依次加入5倍当量的DIPEA，缩合剂HATU(1.05个当量)，搅拌15分钟后加入3倍盐酸羟胺，反应完毕，制备型HPLC纯化得最终产物。
表2.合成最终化合物一览表
表3.化合物抗菌活性一览表
表4.化合物在30μM的浓度条件下抗结直肠癌细胞增殖抑制率活性
结果表明：实施例6、22、29、36、38、40、46、51、57、62、63、64、65、66、70、72、73、75、76、78、79中有21个化合物在30μM的浓度下对结直肠癌细胞HCT116的增殖抑制率超过了50％，其中实施例29化合物的增殖抑制率最高，为94％。说明所合成的化合物在30μM的浓度下具有明显的抑制结直肠癌细胞HCT116增殖的作用。
表5.优选化合物在30μM的浓度条件下抗结直肠癌细胞、肺癌细胞、胃癌细胞、肝癌细胞等增殖抑制率活性
选择实施例6、22、25、29、32、33、34、36、38、40、46、51、55、57、61、62、63、64、65、66、67、70、71、72、73、75、76、77、78、79中，对结直肠癌细胞HCT116增殖抑制效果好(抑制率≥30％)的30个化合物，考察它们对其它癌症细胞如：肺癌细胞、胃癌细胞、肝癌细胞等增殖抑制率活性。结果表明：实施例46、57、62、63、64、65、66、70、72、73、76、78、79中13个化合物在30μM
的浓度下对肺癌细胞A549的增殖抑制率超过了50％，其中实施例76化合物的增殖抑制率最高，为94％；实施例6、29、38、40、62、63、64、65、66、72、73、75、76、79中14个化合物在30μM的浓度下对胃癌细胞MGC-803的增殖抑制率超过了50％，其中实施例29化合物的增殖抑制率最高，为78％；实施例6、22、25、29、33、34、36、38、40、46、51、55、57、62、63、64、65、66、67、70、72、73、75、76、77、78、79中27个化合物在30μM的浓度下对肝癌细胞BEL-7402的增殖抑制率超过了50％，其中实施例64、65、73化合物的增殖抑制率最高，为81％。
表6.优选的化合物抗结直肠癌细胞、肺癌细胞、胃癌细胞、肝癌细胞的IC50值(μM)
表7.实施例64对不同肿瘤细胞的生长抑制活性(IC50，μM)
| 细胞组织来源 | 细胞名称 | IC50(μM) |
| 直肠癌 | HCT-116 | 4.88 |
| T细胞白血病 | Jurkat | 7.61 |
| 急性髓性白血病 | HL60 | 1.21 |
| 肝癌 | BEL7402 | 5.00 |
| 肺癌 | A549 | 4.86 |
| *** | Hela | 11.45 |
| 胃癌 | MGC-803 | 6.11 |
| 乳腺癌 | MCF-7 | 6.62 |
| ***癌 | PC3 | 6.17 |
| 骨肉瘤 | SAOS-2 | 2 |
实施例1
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己酰胺基)-N-(1H-吡唑-3-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1:在60mL二氧六环中加入3-氨基吡唑(1.50g,18.0mmol)，三甲胺(4.5g,20.6mmol)，4-(二甲氨基)吡啶(0.15g,1.2mmol)，搅拌溶解后加入Boc2O，加热回流8h，反应完毕，旋出溶剂，然后用EA稀释并萃取，依次用10％的柠檬酸、饱和食盐水洗涤，有机相浓缩后得油状物，过柱(PE/DCM＝2/1)得白色固体状产物(1.6g,产率48％)。
步骤2:操作如合成通式(X1)中的步骤1。在酸(2.05g,8.5mmol)中加入20mLDMF溶解后，冰浴下加入N-甲基咪唑(1.54g,18.7mmol)，然后缓慢滴加MsCl(1.07g,9.4mmol)，搅拌15min后加入Boc保护的胺(1.56g,8.5mmol)，反应用TLC检测，反应完毕，EA稀释，用10％的柠檬酸洗涤，水相用50mL EA萃取2次，合并有机相后依次用饱和碳酸氢钠、饱和食盐水洗涤，有机相浓缩得油状产物。
步骤3:向油状物中加入20mL和10mL的5M的HCl/MeOH溶液，室温下反应过夜，反应完毕，旋干得油状物(2.0g,两步产率100％)。
1H NMR(400MHz,DMSO)δ11.53(s,1H),10.66(d,J＝4.0Hz,1H),8.94(s,1H),7.79(s,1H),6.52(s,1H),4.70-4.51(m,1H),3.31-3.24(m,1H),3.21-3.15(m,1H),2.43-2.27(m,1H),2.01(dd,J＝12.7,7.5Hz,1H),0.80-0.57(m,4H).
13C NMR(101MHz,DMSO)δ165.97,144.77,96.15,55.94,51.54,37.61,18.42,10.22,9.83.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5：操作如合成通式(X2)中的步骤4。两步收率为18％。
LC-MS(ESI):[M+1]+＝378.15,tR＝1.88min.
1H NMR(400MHz,CDCl3)δ8.36-7.98(m,1H),7.80-7.75(m,1H),6.00-5.84(m,1H),5.67-5.41(m,1H),5.15(s,1H),4.40-3.87(m,2H),3.86-3.31(m,2H),3.26-2.72(m,1H),2.44-2.36(m,1H),2.07-1.83(m,1H),1.84-1.65(m,1H),1.67-1.21(m,5H),0.97-0.82(m,3H),0.76-0.42(m,4H).
13C NMR(101MHz,CDCl3)δ172.19,167.99,158.69,157.56,129.84,101.83,58.87,54.75,51.31,40.60,37.93,29.81,28.93,22.74,20.77,13.96,12.89,8.80.
HRMS(ESI):calculated for C18H27N5O4Na[M+Na]+＝400.1961；found 400.1949.
实施例2
5-氟-2-((S)-5-((R)-2-((N-羟基甲酰胺基)甲基)己羰基)-5-氮杂螺[2.4]庚烷-6-酰胺基)吡啶N-氮氧化合物的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2(2.1g,白色固体，两步收率68％)。
步骤3：操作如合成通式(X2)中的步骤3。
步骤4：所得油状物(242mg,0.46mmol)溶于乙酸乙酯(3mL)，加入过氧化氢尿素复合物(133mg,1.40mmol)和邻苯二甲酸酐(207mg，1.40mmol)。混合物
在室温搅拌2小时。反应结束后，用硫代硫酸钠淬灭反应，用乙酸乙酯萃取。干燥浓缩有机相得到粗品。
步骤5：操作如合成通式(X2)中的步骤4。三步收率为45％。
LC-MS(ESI):[M+1]+＝423.02,tR＝1.80min.
1H NMR(400MHz,CDCl3)δ10.55(s,1H),8.49(dd,J＝9.4,6.6Hz,1H),8.21-8.18(m,1H),7.79(s,1H),7.19-7.09(m,1H),4.97(dd,J＝8.5,3.6Hz,1H),4.00-3.78(m,2H),3.50-3.26(m,2H),3.17-2.80(m,1H),2.40-2.25(m,1H),2.11-1.92(m,1H),1.80-1.46(m,2H),1.44-1.26(m,4H),0.89(t,J＝6.8Hz,3H),0.76-0.61(m,4H).
13C NMR(101MHz,CDCl3)δ173.35,170.54,157.91,154.73(d,J＝248.1Hz),141.69,127.21(d,J＝36.5Hz),115.69(d,J＝20.1Hz),114.77(d,J＝7.8Hz),61.81,54.81,51.31,40.55,36.87,29.81,28.96,22.67,21.25,13.87,12.94,8.46.
HRMS(ESI):calculated for C20H27N4O5NaF[M+Na]+＝445.1863；found 445.1845.
实施例3
(S)-N-(5-(特丁基)异恶唑-3-基)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2(1.4g,白色固体，两步收率47％)。
1H NMR(400MHz,D2O)δ6.14(s,1H),4.44-4.41(m,1H),3.12-2.93(m,2H),2.19(dd,J＝13.4,8.9Hz,1H),1.86(dd,J＝13.4,6.1Hz,1H),0.52-0.29(m,4H).
13C NMR(101MHz,D2O)δ183.22,167.90,157.06,93.24,60.23,52.67,37.13,32.51,27.67,20.09,9.86,8.49.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率28％。
LC-MS(ESI):[M+1]+＝435.24,tR＝2.25min.
1H NMR(400MHz,CDCl3)δ9.94(s,1H),7.81(s,1H),6.69(s,1H),4.84(dd,J＝8.2,4.2Hz,1H),4.07-3.68(m,2H),3.68-3.30(m,2H),3.20-2.80(m,1H),2.36-1.95(m,2H),1.74-1.43(m,2H),1.39-1.23(m,13H),0.91-0.82(m,3H),0.76-0.60(m,4H).
13C NMR(101MHz,CDCl3)δ181.38,173.47,169.95,157.67,157.62,93.55,61.29,55.06,51.36,40.77,36.21,32.98,29.85,28.92,28.64,22.70,21.28,13.85,12.41,9.18.
HRMS(ESI):calculated for C22H34N4O5Na[M+Na]+＝457.2427；found 457.2426.
实施例4
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(4-甲基-3-((4-(吡啶-3-基)嘧啶-2-基)氨基)苯基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2(3.1g,黄色固体，两步收率66％)。
1H NMR(400MHz,D2O)δ9.39(s,1H),9.10(d,J＝8.1Hz,1H),8.87(d,J＝5.5Hz,1H),8.39(d,J＝6.1Hz,1H),8.16-8.11(m,1H),7.63(s,1H),7.55(d,J＝6.2Hz,1H),7.25(d,J＝8.2Hz,1H),7.16(d,J＝7.9Hz,1H),4.66-4.60(m,1H),3.27(d,J＝3.3Hz,2H),2.39(dd,J＝13.1,8.9Hz,1H),2.12(s,3H),2.10-2.00(m,1H),0.73-0.57(m,4H).13C NMR(101MHz,D2O)δ167.92,164.58,155.40,151.87,145.43,143.57,141.24,135.16,134.34,133.74,131.92,131.34,127.79,120.66,118.45,108.22,60.22,52.71,37.45,20.18,16.50,9.61,8.85.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得淡黄色固体，两步收率38％。
LC-MS(ESI):[M+1]+＝572.27,tR＝2.11min.
1H NMR(400MHz,CDCl3)δ9.38(s,1H),9.24(s,1H),8.67(d,J＝4.0Hz,1H),8.51-8.25(m,3H),7.87(s,1H),7.49-7.39(m,1H),7.38-7.05(m,4H),5.06-4.61(m,1H),4.13-3.61(m,2H),3.57-3.22(m,2H),3.19-2.84(m,1H),2.32-2.24(m,3H),2.21-2.09(m,2H),1.85-1.38(m,2H),1.36-1.12(m,4H),0.94-0.69(m,3H),0.71-0.54(m,4H).
13C NMR(101MHz,CDCl3)δ173.98,168.95,162.54,160.62,159.01,157.23,151.09,148.33,137.43,136.89,135.19,132.84,130.57,124.48,123.84,115.34,113.32,108.02,61.63,55.45,51.49,41.14,35.16,30.12,28.89,22.66,21.11,18.40,17.66,13.70,8.32.
HRMS(ESI):calculated for C31H38N7O4[M+H]+＝572.2985；found 572.2980.
实施例5
(S)-N-(3-氟吡啶-2-基)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2(1.7g,白色固体，两步收率45％)。
1H NMR(400MHz,D2O)δ8.20(d,J＝5.5Hz,1H),8.06(t,J＝9.0Hz,1H),7.56-7.50(m,1H),4.90-4.79(m,1H),3.31(s,2H),2.61-2.39(m,1H),2.15(dd,J＝13.4,6.1Hz,1H),0.76-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.94,150.12(d,J＝255.5Hz),138.18,137.92(d,J＝13.4Hz),130.12(d,J＝16.9Hz),123.10(d,J＝5.8Hz),60.39,52.83,37.21,20.06,9.96,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到淡黄色固体，两步收率37％。
LC-MS(ESI):[M+1]+＝406.70,tR＝2.35min.
1H NMR(400MHz,CDCl3)δ8.40-8.15(m,1H),8.01-7.79(m,1H),7.65-7.35(m,1H),7.33-7.00(m,1H),5.05-4.66(m,1H),4.30-3.64(m,2H),3.62-3.21(m,2H),3.05(m,1H),2.78-1.90(m,2H),1.84-1.40(m,2H),1.41-1.14(m,4H),1.00-0.51(m,7H).
13C NMR(101MHz,CDCl3)δ173.98,168.98,157.19,150.39(d,J＝258.5Hz),143.63(d,J＝5.6Hz),140.45(d,J＝12.1Hz),123.73(d,J＝17.4Hz),121.04,61.31,55.35,51.28,40.97,35.11,30.08,28.90,22.67,21.10,13.85,13.71,8.13.
HRMS(ESI):calculated for C20H28FN4O4[M+H]+＝407.2110；found 407.2126.
实施例6
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(5-甲基噻唑-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2(680mg,白色固体，两步收率100％)。
1H NMR(400MHz,D2O)δ7.24(s,1H),4.77(dd,J＝8.9,6.5Hz,1H),3.24(s,2H),2.38(dd,J＝13.4,9.1Hz,1H),2.28(s,3H),2.05(dd,J＝13.4,6.3Hz,1H),0.72-0.45(m,4H).
13C NMR(101MHz,D2O)δ168.01,159.21,129.86,124.06,60.16,52.75,36.68,20.02,11.03,9.78,8.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到类白色固体，两步收率42％。
LC-MS(ESI):[M+1]+＝408.72,tR＝2.67min.
1H NMR(400MHz,CDCl3)δ7.70(s,1H),7.64(s,1H),4.68(d,J＝7.6Hz,1H),4.24(t,J＝10.5Hz,1H),3.93(d,J＝11.5Hz,1H),3.41(d,J＝12.5Hz,1H),3.02(d,J＝11.5Hz,1H),2.62(s,1H),2.40(dd,J＝12.2,8.0Hz,1H),2.30(s,3H),2.13(d,J＝12.3Hz,
1H),1.73-1.41(m,2H),1.39-1.16(m,4H),0.88(t,J＝6.6Hz,3H),0.72-0.50(m,2H),0.30-0.05(m,2H).
13C NMR(101MHz,CDCl3)δ174.04,171.12,158.73,153.63,135.01,126.02,61.53,53.99,53.60,43.50,39.25,31.29,29.17,22.90,19.23,13.84,13.26,11.72,5.98.
HRMS(ESI):calculated for C19H29N4O4S[M+H]+＝409.1910；found 409.1922.
实施例7
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(3-(吡啶-3-基)苯基)-5-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：在混合物3-溴吡啶(2.37g,15mmol)、3-氨基苯硼酸单水合物(2.32g,15mmol)和Pd(PPh3)4(520mg,0.45mmol)中加入乙二醇二甲醚(30mL)和水(15mL)，体系用氮气置换三次，加热回流12小时。反应结束后，冷却，除去溶剂，剩余物经过柱分离(DCM/EA：5/1到2/1)得到黄色半固体产物(680mg,27％收率)。
1H NMR(400MHz,CDCl3)δ8.81(d,J＝2.0Hz,1H),8.57(,J＝4.7Hz,1H),7.84(d,J＝7.9Hz,1H),7.34(dd,J＝7.9,4.8Hz,1H),7.26(dd,J＝8.8,6.8Hz,1H),6.96(d,J＝7.6Hz,1H),6.88(s,1H),6.73(dd,J＝8.0,2.1Hz,1H),3.83(brs,2H).
13C NMR(101MHz,CDCl3)δ148.31,148.21,147.06,138.96,136.88,134.41,130.05,123.52,117.48,114.82,113.63.
步骤2：操作如合成通式(X1)中的步骤1。将(S)-5-(叔丁基)-5-氮杂螺[2.4]庚烷-6-羧酸(950mg,3.94mmol)溶于DMF(5mL)中，然后加入NMI(0.7mL,8.67mmol)。冷至0℃，滴加MsCl(451mg,3.94mmol)，搅拌15分钟。然后加入3-(吡啶-3-基)苯胺(670mg,3.94mmol)。TLC监控反应进程。待反应结束后，反应液用乙酸乙酯稀释，10％柠檬酸水溶液洗涤，水相用乙酸乙酯萃取(2x20mL)。有机相合并后先用饱和碳酸钠水溶液洗，然后用饱和食盐水洗。于旋蒸下减压浓缩后得粗产物。
步骤3:将粗品溶于10mL***中，加入5mL HCl/MeOH溶液(5M)，室温搅拌过夜。收集并干燥沉淀得棕色固体(900mg,两步69％收率)。
1H NMR(400MHz,D2O)δ8.54(s,1H),8.35(d,J＝5.7Hz,1H),8.31(d,J＝8.2Hz,1H),7.69(dd,J＝7.8,6.2Hz,1H),7.38(s,1H),7.15-7.05(m,2H),4.44-4.38(m,1H),3.08(d,J＝11.4Hz,1H),3.03(d,J＝11.4Hz,1H),2.16(dd,J＝13.2,8.8Hz,1H),1.80(dd,J＝13.3,6.6Hz,1H),0.54-0.29(m,4H).
13C NMR(101MHz,D2O)δ167.95,144.27,139.57,138.97,138.77,137.47,133.84,130.50,127.45,124.14,122.41,119.26,60.25,52.75,37.49,20.24,9.55,9.03.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到类白色固体，两步收率19％。
LC-MS(ESI):[M+1]+＝464.85,tR＝2.33min.
1H NMR(400MHz,CDCl3)δ9.68(s,1H),8.65(d,J＝1.6Hz,1H),8.43(d,J＝3.8Hz,1H),7.84-7.71(m,2H),7.66(s,1H),7.39(d,J＝7.6Hz,1H),7.23(dd,J＝7.8,4.7Hz,1H),7.16(t,J＝7.9Hz,1H),7.05(d,J＝7.6Hz,1H),4.81(t,5.2Hz),3.92-3.58(m,2H),3.56-3.32(m,2H),3.15-3.02(m,1H),2.14-1.98(m,1H),1.72-1.34(m,2H),1.36-1.11(m,4H),0.80-0.64(m,3H),0.63-0.48(s,4H).
13C NMR(101MHz,CDCl3)δ173.77,169.72,157.20,147.90,147.76,139.28,137.91,136.55,134.85,129.36,123.65,122.19,119.26,118.10,61.64,55.59,51.68,50.53,41.17,36.00,30.12,28.87,22.66,21.12,13.73,9.05.
HRMS(ESI):calculated for C26H32N4O4Na[M+Na]+＝487.2321；found 487.2318.
实施例8
(S)-5-((R)-2-((N-羟酰胺基)甲基)己羰基)-N-(3-(2-吡啶氨基)苯基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1:在反应瓶中加入xantphos(347mg,3mol％)、Pd2(dba)3(366mg,2mol％)2-氨基嘧啶(1.92g,20mmol)和叔丁醇钠(1.92g,20mmol)。将体系用氮气置换三次。3-溴-1-硝基苯(4.04g,20mmol)的甲苯溶液(20mL)加入到反应中。加热至95℃，反应18小时。冷却后，硅藻土过滤(滤饼先用200mL甲苯洗以除去杂质，而后用10％MeOH的乙酸乙酯混合液(500mL)洗脱得粗品，经乙酸乙酯打浆得黄色固体纯品(2.6g,60％收率)。
LC-MS(ESI):[M+1]+＝217.00,tR＝2.45min.
1H NMR(400MHz,CDCl3)δ8.74(s,1H),8.51(d,J＝4.7Hz,2H),7.90-7.83(m,2H),7.70(s,1H),7.47(t,J＝8.1Hz,1H),6.85(t,J＝4.7Hz,1H).
13C NMR(101MHz,CDCl3)δ159.57,158.12,148.86,140.74,129.51,124.47,116.94,113.74,113.61.
步骤2:将含10％Pd/C(50mg)，N-(3-硝基苯基)嘧啶基-2-胺(1.3g,6.0mmol)80％水合肼(1mL)的三氟乙醇(20mL)溶液，加热到80℃反应5小时。反应结束后，过滤掉催化剂，所得滤液浓缩可得产物(1.1g,100％收率)。
LC-MS(ESI):[M+1]+＝186.76,tR＝1.17min.
1H NMR(400MHz,CDCl3)δ8.39(d,J＝4.8Hz,2H),7.86(brs,1H),7.16(t,J＝1.9Hz,1H),7.09(t,J＝8.0Hz,1H),6.94-6.83(m,1H),6.66(t,J＝4.8Hz,1H),6.37(dd,J＝7.9,2.0Hz,1H),3.67(brs,2H).
13C NMR(101MHz,CDCl3)δ160.29,157.96,147.26,140.52,129.71,112.29,109.95,109.75,106.28.
步骤3:操作如合成通式(X1)中的步骤1。(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-羧酸(965mg,4.0mmol)溶于DMF(8mL)，加入NMI(722mg,8.8mmol)冷至0℃,滴加MsCl(460mg,4.0mmol)，搅拌15分钟。而后加入N-(2-嘧啶基)苯-1,3-二胺(745mg,4.0mmol)。反应结束后，用乙酸乙酯稀释，10％柠檬酸水溶液洗涤，有机相用乙酸乙酯萃取(2x50mL)。合并的有机相用饱和碳酸钠和食盐水洗涤。减压浓缩得黄色固体。
步骤4:固体溶于2:1DCM/TFA(8mL/4mL)溶液中，在室温下反应1小时。反应完毕后加入几滴水，用固体碳酸钠固体中和。经无水Na2SO4干燥，滤除不溶固体后浓缩滤液得游离碱(500mg,两步收率50％)。
1H NMR(400MHz,CDCl3)δ9.81(brs,1H),8.34(m,2H),8.00-7.90(m,1H),7.72-7.50(m,1H),7.46-7.25(m,1H),7.26-7.09(m,1H),6.71-6.47(m,1H),4.19-3.76(m,1H),3.16-2.85(m,1H),2.91-2.52(m,1H),2.28-2.05(m,1H),2.00-1.77(m,1H),0.82-0.21(m,4H).
13C NMR(101MHz,CDCl3)δ172.97,160.08,157.97,140.14,138.46,129.36,115.19,113.66,112.55,110.39,61.41,54.68,39.12,22.40,10.88,9.55.
步骤5：操作如合成通式(X2)中的步骤3。
步骤6:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到类白色固体，两步收率37％。
1H NMR(400MHz,CDCl3)δ9.81(brs,1H),8.34(m,2H),8.00-7.90(m,1H),7.72-7.50(m,1H),7.46-7.25(m,1H),7.26-7.09(m,1H),6.71-6.47(m,1H),4.19-3.76(m,1H),3.16-2.85(m,1H),2.91-2.52(m,1H),2.28-2.05(m,1H),2.00-1.77(m,1H),0.82-0.21(m,4H).
13C NMR(101MHz,CDCl3)δ172.97,160.08,157.97,140.14,138.46,129.36,115.19,113.66,112.55,110.39,61.41,54.68,39.12,22.40,10.88,9.55.
LC-MS(ESI):[M+1]+＝481.36,tR＝2.35min.
1H NMR(400MHz,CDCl3)δ9.30(s,1H),8.37(d,J＝4.7Hz,2H),8.12(s,1H),7.87(s,1H),7.77(s,1H),7.47(d,J＝7.5Hz,1H),7.24-7.07(m,2H),6.66(t,J＝4.8Hz,1H),5.06-4.47(m,1H),4.08-3.56(m,2H),3.55-3.30(m,2H),3.23-2.76(m,1H),2.29-1.88(m,2H),1.75-1.41(m,2H),1.40-1.19(m,4H),1.05-0.70(m,3H),0.74-0.45(m,4H).
13C NMR(101MHz,CDCl3)δ173.78,169.36,159.92,157.93,157.38,139.95,138.86,129.19,115.26,114.12,112.26,110.94,61.66,55.48,51.60,41.14,35.63,30.14,28.92,22.68,21.13,13.78,12.95,8.90.
HRMS(ESI):calculated for C25H33N6O4[M+1]+＝481.2563；found 481.2566.
实施例9
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(4-(3-吡啶基)嘧啶-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1:将3-二甲氨基-1-(3-吡啶基)-2-丙烯基-1-酮(1.76g,10.0mmol)悬于正丁醇(20mL)，依次加入盐酸胍(1.15g,12.0mmol)和NaOH(480mg,12.0mmol)。混合物加热到120℃，反应过夜。收集析出的固体，用水(50mL)洗涤后，真空干燥得淡黄色晶体(1.51g,88％收率)。
1H NMR(400MHz,DMSO)δ9.23(d,J＝2.0Hz,1H),8.67(dd,J＝4.7,1.3Hz,1H),8.38(dt,J＝8.0,1.8Hz,1H),8.36(d,J＝5.1Hz,1H),7.52(dd,J＝8.0,4.8Hz,1H),7.20(d,J＝5.1Hz,1H),6.81(s,2H).
13C NMR(101MHz,DMSO)δ163.78,161.53,159.36,151.11,147.93,134.11,132.44,123.72,105.99.
步骤2:将(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-羧酸(2.0g,8.5mmo)溶于DMF(20mL)，加入NMI(2.1g,25.5mmol)，冷至0℃，滴加MsCl(978mg,8.5mmol)，搅拌15分钟。然后加入4-(3-吡啶基)嘧啶-2-胺(980mg,5.7mmol)和氯化锂(721mg,17.0mmol)。室温搅拌反应48小时后，反应液用乙酸乙酯稀释，用10％柠檬酸水溶液洗涤，水相用乙酸乙酯萃取(2x50mL)。有机相用饱和碳酸钠和食盐水洗。减压浓缩后，粗品用硅胶柱纯化(DCM/MeOH＝100/1-30/1)得纯品(1.0g,44％收率)。
1H NMR(400MHz,CDCl3)(two rotomers were observed)δ9.98(brs,0.5H),9.28(d,J＝2.0Hz,1H),9.06(brs,0.5H),8.83-8.70(m,2H),8.42(d,J＝8.0Hz,1H),7.56-7.40(m,2H),4.96-4.60(m,1H),3.65-3.50(m,1H),3.45-3.10m,1H),2.50-2.25(m,1H),2.20-2.05(m,1H),1.51(s,9H),0.78-0.54(s,4H).
步骤3:分离所得的产物溶于10mL***中，加入5mL HCl/MeOH溶液(5M)，室温搅拌反应过夜。收集沉淀，干燥后得白色固体(584mg,63％收率)。
1H NMR(400MHz,D2O)δ9.09(s,1H),8.65(d,J＝5.4Hz,1H),8.62(d,J＝3.9Hz,1H),8.47(d,J＝8.1Hz,1H),7.68(d,J＝5.3Hz,1H),7.61(dd,J＝8.0,5.2Hz,1H),4.79-4.76(m,1H),3.42-3.27(m,2H),2.50(dd,J＝13.4,9.0Hz,1H),2.15(dd,J＝13.2,6.1Hz,1H),0.94-0.60(m,4H).
13C NMR(101MHz,D2O)δ162.87,159.54,149.77,146.43,138.16,137.44,134.43,132.17,124.98,114.33,60.83,52.79,37.24,26.35,20.10,18.44,9.85,8.57.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到类白色固体，两步收率19％。
LC-MS(ESI):[M+1]+＝466.74,tR＝2.20min.
1H NMR(400MHz,CDCl3)δ9.91(s,1H),9.37-9.08(m,1H),8.85-8.53(m,1H),8.37(s,2H),7.87(s,1H),7.65-7.12(m,2H),5.55-5.00(m,1H),4.14-3.64(m,2H),3.55-3.10(m,2H),3.02-2.58(m,1H),2.45-2.10(m,2H),2.05-1.46(m,2H),1.45-1.10(m,4H),1.00-0.30(m,7H).
13C NMR(101MHz,CDCl3)δ180.38,173.69,162.72,159.32,157.91,157.76,151.70,148.31,134.89,131.89,123.83,111.79,61.70,55.39,51.35,42.84,41.02,35.71,30.09,28.87,22.71,21.02,13.75,8.41.
HRMS(ESI):calculated for C24H31N6O4[M+1]+＝467.2407；found 467.2399.
实施例10
N-((R)-2-((S)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-羰基)己基)-N-羟甲酰胺的合成
步骤1:将(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-甲酸(2.4g,10mmol)放在烧瓶中，加入N,N-二甲基甲酰胺DMF(25mL)使之溶解，冰浴下加入N-甲基咪唑NMI(1.8mL,22mmol)，然后滴加甲磺酰氯(0.78mL,10mmol)，0℃下搅拌15分钟。然后加入邻苯二胺(2.8g,20mmol)。反应体系于室温下搅拌6小时。待反应结束，反应液用乙酸乙酯稀释，有机相用10％柠檬酸水溶液洗涤三次除去过量的邻苯二胺。最后有机相经干燥、浓缩后得粉红色粗品，为泡沫状固体。
步骤2:将所得固体溶解于20mL甲基叔丁基醚中，加入4mL醋酸。将该溶液加热回流3小时，反应结束后用饱和Na2CO3溶液中和至中性。分离得到有机层，浓缩得到关环产物。
步骤3:将油状关环产物溶解在20mL***中，加入5M的HCl/MeOH溶液(10mL)，搅拌过夜。收集析出的白色固体并真空干燥之，得1.3g产品，三步总收率45％。
1H NMR(400MHz,D2O)δ7.67-7.58(m,2H),7.49-7.39(m,2H),5.41(t,J＝8.6Hz,1H),3.55-3.46(m,1H),3.25-3.15(m,1H),2.68-2.57(m,1H),2.39-2.28(m,1H),0.85-0.60(m,4H).
13C NMR(101MHz,D2O)δ144.90,130.95,127.05,114.21,36.52,20.20,11.92,7.25.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到半固体，两步收率为32％。
LC-MS(ESI):[M+1]+＝385.06,tR＝1.51min.
1H NMR(400MHz,CDCl3)δ12.10(s,1H),8.30-7.79(m,1H),7.74-7.34(m,2H),7.29-7.16(m,2H),5.58-5.42(m,1H),4.21-4.01(m,1H),3.97-3.87(m,1H),3.84-3.68(m,1H),3.55-3.40(m,1H),3.31-3.11(m,1H),2.76-2.57(m,1H),2.40-2.07(m,1H),1.70-1.45(m,2H),1.39-1.22(m,4H),0.94-0.83(m,3H),0.73-0.49(m,2H),0.32-0.15(m,2H).
13C NMR(101MHz,CDCl3)δ174.59,161.90,154.44,123.20,122.92,56.75,53.81,48.09,43.15,41.24,29.69,29.11,22.86,19.00,14.11,13.86,5.90.
HRMS(ESI):calculated for C21H28N4O3[M+1]+＝385.2060；found 385.2013.
实施例11
N-羟基-N-((R)-2-((S)-6-(3-(吡啶-3-基)-1,2,4-恶二唑-5-基)-5-氮杂螺[2.4]庚烷-5-羰基)己基)甲酰胺的合成
步骤1:将3-氰基吡啶(5.2g,50mmol)溶于50mL乙醇中,然后加入羟胺盐酸盐(4.2g,60mmol),NaHCO3(5.0g,60mmol)和水(3mL)。混合物加热回流12小时。加入无水Na2SO4，过滤后用***(50mL)稀释后析出白色晶体(6.6g,96％收率)。
1H NMR(400MHz,DMSO)δ9.87(s,1H),8.87(d,J＝2.1Hz,1H),8.57(dd,J＝4.8,1.5Hz,1H),8.03(dt,J＝8.0,1.9Hz,1H),7.41(dd,J＝8.0,4.8Hz,1H),6.01(s,2H).
13C NMR(101MHz,DMSO)δ149.72,148.92,146.56,132.82,129.01,123.22.
步骤2:将(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-羧酸(2.49g,10.3mmol)溶于DMF(25mL)中，然后加入N,N'-羰基二咪唑(CDI,1.75g,10.3mmol)。混合物室温搅拌8小时后，加入N-羟基-(3-吡啶基)甲脒(1.41g,10.3mmol)，然后再加入另一当量的CDI(1.75g,10.3mmol)。于氮氛下，将反应加热到90℃，反应12小时。冷却后，用乙酸乙酯(50mL)稀释，经10％柠檬酸水溶液洗涤，乙酸乙酯(50mL)萃取。收集的有机相依次用10％柠檬酸水溶液,饱和碳酸氢钠水溶液和食盐水洗涤，浓缩后得粗品(3.2g,84％收率)。
LC-MS(ESI):[M+1]+＝343.19,tR＝2.32min.
1H NMR(400MHz,CDCl3)(two rotomers were observed)δ9.35-9.29(m,1H),8.79-8.73(m,1H),8.56-8.27(m,1H),7.55-7.37(m,1H),5.38-5.20(m,1H),3.58-3.38(m,2H),2.64-2.41(m,1H),1.47(s,4H),1.32(s,5H),0.81-0.47(m,4H).
步骤3:油状物(2.1g,5.67mmol)溶于10mL***，加入10mL HCl/MeOH溶液(5M)，反应搅拌过夜。反应液加少量水，加碳酸钠固体中和酸，待气泡消失后，滤除固体，滤液浓缩干后即得游离碱(1.2g,80％收率)。
LC-MS(ESI):[M+1]+＝243.89,tR＝1.13min.
1H NMR(400MHz,CDCl3)δ9.33(d,J＝1.7Hz,1H),8.74(dd,J＝4.8,1.4Hz,1H),8.37(dt,J＝7.9,1.7Hz,1H),7.43(dd,J＝7.9,4.9Hz,1H),4.76(dd,J＝7.7,6.1Hz,1H),3.14(d,J＝10.0Hz,1H),3.01(d,J＝10.0Hz,1H),2.30(dd,J＝12.6,7.8Hz,1H),2.19(dd,J＝12.6,5.9Hz,1H),0.65(s,4H).
13C NMR(101MHz,CDCl3)δ182.26,166.29,151.99,148.70,134.72,123.59,123.11,55.07,54.87,40.42,22.22,11.54,11.38.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到无色油状物，两步收率33％。
LC-MS(ESI):[M+1]+＝413.69,tR＝2.33min.
1H NMR(400MHz,CDCl3)δ9.20(d,J＝1.2Hz,1H),8.79-8.57(dd,J＝3.6Hz,1H),8.28(dd,J＝6.2,1.6Hz,1H),7.73(s,1H),7.35(dd,J＝7.9,4.9Hz,1H),5.51(dd,J＝8.1,4.0Hz,1H),3.94-3.67(m,2H),3.66-3.47(m,1H),3.42-3.26(m,1H),3.16-2.82(m,
1H),2.40(dd,J＝12.9,8.2Hz,1H),2.03-1.78(m,1H),1.64-1.39(m,2H),1.37-1.16(m,4H),0.84(t,J＝7.0Hz,3H),0.74-0.42(m,4H).
13C NMR(101MHz,CDCl3)δ180.16,172.50,166.33,157.43,151.85,148.55,134.84,123.63,123.28,54.52,54.10,51.66,40.86,38.86,29.99,28.76,22.77,21.17,13.87,12.48,8.58.
HRMS(ESI):calculated for C21H28N5O4[M+1]+＝414.2141；found 414.2146.
实施例12
N-羟基-N-((R)-2-((S)-6-(5-(吡啶-2-基)-1,3,4-恶二唑-2-基)-5-氮杂螺[2.4]庚烷-5-羰基)己基)甲酰胺的合成
步骤1:(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-羧酸(1.0g,4.14mmol)溶于DMF(10mL)中，加入NMI(1.8mL,22mmol)，在0℃下，滴加MsCl(475mg,4.14mmol)，搅拌15分钟。然后加入2-吡啶甲酰肼(569mg,4.14mmo)。反应6小时后，用乙酸乙酯稀释，10％柠檬酸水溶液洗涤，水相用乙酸乙酯萃取(2x50mL)。合并的有机相依次用饱和碳酸氢钠和食盐水洗涤，浓缩后即得无色油状产物(1.5g,100％收率)。
LC-MS(ESI):[M+1]+＝361.09,tR＝2.10min.
步骤2:油状物(1.5g,4.14mmol)溶于THF(20mmol)中，然后分批加入Burgess试剂(2.9g,10.35mmol)，混合物在室温下搅拌3小时。待反应完成后，反应液用***稀释，然后加入水。分离得到的有机相经饱和食盐水洗涤两次，无水Na2SO4干燥，过滤，浓缩后得粗产物(1.4g,100％收率)。
LC-MS(ESI):[M+1]+＝343.13,tR＝2.53min.
步骤3:油状物(1.4g,4.13mmol)溶于2:1DCM/TFA(6mL/3mL)中，混合物室温搅拌反应1小时。反应结束后，反应液中加入几滴水，然后用碳酸钠固体中和。经无水硫酸钠干燥，滤去不溶物。滤液浓缩至干得游离碱(1.0g,100％收率)。
LC-MS(ESI):[M+1]+＝243.08,tR＝1.34min.
1H NMR(400MHz,CDCl3)δ8.78(d,J＝4.6Hz,1H),8.25(d,J＝7.9Hz,1H),7.89(td,J＝7.8,1.6Hz,1H),7.46(ddd,J＝7.6,4.9,0.9Hz,1H),4.78(dd,J＝7.4,6.4Hz,1H),3.14(d,J＝10.0Hz,1H),2.98(d,J＝10.0Hz,1H),2.37-2.15(m,2H),0.74-0.52(m,4H).
13C NMR(101MHz,CDCl3)δ169.54,164.25,150.23,143.57,137.21,125.79,123.10,54.79,54.16,39.87,22.21,11.59,11.32.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到无色油状物，两步收率25％。
LC-MS(ESI):[M+1]+＝413.76,tR＝2.47min.
1H NMR(400MHz,CDCl3)δ8.45(d,J＝4.0Hz,1H),8.19-7.85(m,1H),7.75-7.25(m,2H),7.22-7.00(m,1H),5.50-4.92(m,1H),4.04-3.44(m,2H),3.42-2.96(m,2H),2.98-2.60(m,1H),2.58-2.07(m,1H),2.06-1.57(m,1H),1.45-1.17(m,2H),1.16-0.92(m,4H),0.72-0.53(m,3H),0.47-0.16(m,4H).
13C NMR(101MHz,CDCl3)δ175.27,172.56,167.63,166.49,164.20,163.74,161.76,157.54,150.29,150.15,143.56,141.99,138.38,137.11,126.75,125.69,123.48,123.00,54.59,53.87,53.56,53.09,51.04,47.66,41.11,40.99,38.69,38.64,30.74,30.04,29.13,28.77,22.91,22.74,21.02,19.07,15.71,13.86,13.78,12.41,8.83,8.68,6.36.
HRMS(ESI):calculated for C21H28N5O4[M+1]+＝414.2141；found 414.2157.
实施例13
N-((R)-2-((S)-6-([1,2,4]三氮唑[4,3-a]吡啶-3-基)-5-氮杂螺[2.4]庚烷-5-羰基)己基)-N-羟基酰胺的合成
步骤1:将(S)-5-(叔丁氧羰基)-5-氮杂螺[2.4]庚烷-6-羧酸(2.4g,10mmol)溶于DMF(25mL)，加入NMI(1.8mL,22mmol)。冷却至0℃，滴加MsCl(0.78mL,10mmol)，搅拌15分钟。然后2-吡啶肼(1.1g,10mmol)加入到溶液中。反应6小时，用乙酸乙酯稀释，加入水(100mL)，乙酸乙酯萃取(2x50mL)。有机相用饱和食盐水洗涤。旋蒸浓缩得粗品，***打浆后得类白色固体(2.35g,71％收率)。
LC-MS(ESI):[M+1]+＝332.86,tR＝1.74min.
步骤2:将固体(2.35g,7.1mmol)溶于二氯乙烷DCE(30mL)，加入Lawesson试剂(2.9g,7.1mmol)。于氮氛条件下，反应液加热到80℃，反应3小时。结束后，依次用饱和碳酸钠溶液、10％柠檬酸水溶液和饱和食盐水洗涤。浓缩反应液用硅胶柱纯化(洗脱剂：DCM/MeOH＝50/1到30/1)得无色油状物(2.0g,90％收率)。LC-MS(ESI):[M+1]+＝315.22,tR＝2.19min.
步骤3:油状物(2.0g,6.36mmol)溶于20mL***，加入10mLHCl/MeOH溶液(5M)，室温下搅拌过夜。收集得到的沉淀干燥后得白色固体(820mg,45％收率)。LC-MS(ESI):[M+1]+＝214.79,tR＝1.17min.
1H NMR(400MHz,D2O)δ8.68(d,J＝11.1Hz,1H),8.10-7.95(m,2H),7.55(d,J＝3.7Hz,1H),5.76(dd,J＝6.8Hz,1H),3.61(d,J＝11.4Hz,1H),3.48(d,J＝11.4Hz,1H),2.70-2.53(m,2H),0.95-0.71(m,4H).
13C NMR(101MHz,D2O)δ145.96,143.31,137.51,124.76,118.88,111.94,52.75,52.36,36.80,19.88,10.36,9.63.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。柱层析(DCM:MeOH＝10:1)得到无色油状物，两步收率23％。
LC-MS(ESI):[M+1]+＝386.03,tR＝2.00min.
1H NMR(400MHz,CDCl3)δ8.49-8.35(m,1H),7.85(s,1H),7.77-7.71(m,1H),7.28-7.21(m,1H),7.03-6.76(m,1H),5.79-5.39(m,1H),4.04-3.77(m,2H),3.78-3.37(m,2H),3.24-2.90(m,1H),2.87-2.59(m,1H),2.40-2.16(m,1H),1.62-1.30(m,2H),1.22-0.99(m,4H),0.93-0.47(m,7H).
13C NMR(101MHz,CDCl3)δ172.93,157.21,149.67,147.19,127.20,123.46,116.01,113.42,54.93,51.40,48.34,41.16,39.13,29.91,28.77,22.45,21.39,13.72,11.62,10.36.HRMS(ESI):calculated for C20H27N5O3Na[M+Na]+＝408.2012；found 408.2029.
实施例14
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(4-吗啉苯基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝302.05,tR＝1.06min.
1H NMR(400MHz,DMSO)δ10.82(s,1H),7.52(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),4.55(t,J＝7.9Hz,1H),3.78-3.67(m,4H),3.19(dd,J＝37.9,11.2Hz,2H),3.08-3.02(m,4H),2.33(dd,J＝12.9,8.2Hz,1H),2.01(dd,J＝12.9,7.6Hz,1H),0.78-0.53(m,4H).
13C NMR(101MHz,DMSO)δ165.87,147.72,130.32,120.50,115.35,66.04,59.59,51.66,48.71,37.81,20.35,10.34,9.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率30％。
LC-MS(ESI):[M+1]+＝473.29,tR＝1.79min.
1H NMR(400MHz,CDCl3)δ7.80(s,1H),7.40(d,J＝7.4Hz,2H),6.82(d,J＝6.8Hz,2H),4.83(s,1H),4.17-3.64(m,7H),3.53-3.27(m,2H),3.08(s,5H),2.16(s,2H),1.28(d,J＝20.0Hz,6H),0.84(s,3H),0.80-0.50(m,4H).
实施例15
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(吡嗪-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.57min.
1H NMR(400MHz,D2O)δ8.98(d,J＝96.9Hz,1H),8.31(t,J＝81.8Hz,2H),4.66-4.49(m,1H),3.26-2.81(m,2H),2.29-1.62(m,2H),0.63-0.09(m,4H).
13C NMR(101MHz,D2O)δ148.55,144.82,137.12,133.80,60.43,52.76,37.11,20.13,9.77,8.95,8.64.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率26％。
LC-MS(ESI):[M+1]+＝390.19,tR＝1.56min.
1H NMR(400MHz,CDCl3)δ8.48-8.12(m,3H),7.82(s,1H),5.31(s,1H),4.96(s 1H),3.89(d,J＝9.2Hz,1H),3.76-2.54(m,4H),2.30-2.20(m,1H),1.77-1.28(m,7H),0.86(s,3H),0.81-0.58(m,4H).
实施例16
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己酰基)-N-(嘧啶-4-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.43min.
1H NMR(400MHz,D2O)δ9.08(s,1H),8.78(d,J＝6.8Hz,1H),8.42(t,J＝31.2Hz,1H),4.82(dd,J＝8.7,6.4Hz,1H),3.50-3.15(m,2H),2.57-2.00(m,2H),0.88-0.58(m,4H).
13C NMR(101MHz,D2O)δ169.77,152.74,150.17,111.15,60.98,52.82,36.76,20.08,9.91,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率28％。
LC-MS(ESI):[M+1]+＝390.19,tR＝1.76min.
1H NMR(400MHz,CDCl3)δ9.84(d,J＝23.5Hz,2H),9.05-6.75(m,4H),4.97(t,J＝36.3Hz,1H),4.27-2.60(m,5H),1.58-0.98(m,8H),0.97-0.30(m,7H).
实施例17
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(异恶唑-5-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.11,tR＝0.57min.
1H NMR(400MHz,DMSO)δ10.66(s,1H),9.01(s,1H),8.51(d,J＝1.6Hz,1H),6.29(d,J＝1.6Hz,1H),4.98-4.32(m,1H),3.29-2.98(m,2H),2.45-1.87(m,2H),0.85-0.47(m,4H).
13C NMR(101MHz,DMSO)δ165.75,160.13,152.15,88.25,59.47,51.64,37.35,20.12,10.27,9.78
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率27％。
LC-MS(ESI):[M+1]+＝379.17,tR＝1.88min.
1H NMR(400MHz,CDCl3)δ8.14(s,1H),7.78(d,J＝23.1Hz,1H),6.38(d,J＝41.7Hz,1H),5.03-4.72(m,1H),3.97-3.79(m,2H),3.54-2.85(m,3H),2.11(d,J＝12.2Hz,1H),1.74-1.17(m,7H),0.88(s,3H),0.78-0.60(m,4H).
实施例18
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)己酰基)-N-环丙基-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝182.98,＝0.55min
1H NMR(400MHz,CDCl3)δ7.82(s,1H),3.93(dd,J＝8.7,5.2Hz,1H),3.00-2.95(m,1H),2.89(s,1H),2.77-2.71(m,2H),2.24-1.78(m,2H),0.97-0.25(m,8H).
13C NMR(101MHz,CDCl3)δ175.92,60.94,54.70,39.27,22.37,22.09,11.00,9.48,6.30,6.27.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率28％。
LC-MS(ESI):[M+1]+＝352.14,tR＝1.46min
1H NMR(400MHz,CDCl3)δ8.35(d,J＝16.0Hz,1H),8.02-7.48(m,1H),7.10-6.43(m,1H),4.84-4.32(m,1H),4.02-3.80(m,1H),3.77-3.41(m,2H),3.18-3.02(m,1H),3.01-2.38(m,2H),2.27-1.90(m,2H),1.45-1.05(m,6H),0.99-0.78(m,4H),0.76-0.35(m,7H).
实施例19
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己酰基)-N-(4-甲基噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝237.60,tR＝0.66min.
1H NMR(400MHz,DMSO)δ9.09(d,J＝4.8Hz,1H),6.89(d,J＝0.9Hz,1H),4.92-4.46(m,1H),3.42-2.98(m,2H),2.48-1.85(m,5H),0.88-0.41(m,4H).
13C NMR(101MHz,DMSO)δ167.14,157.01,146.07,108.59,59.22,51.52,37.47,20.13,16.53,10.22,9.90.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率28％。
LC-MS(ESI):[M+1]+＝409.53,tR＝1.90min.
1H NMR(400MHz,CDCl3)δ11.53(s,1H),7.84(S,1H),6.86(s,1H),5.31(s,1H),5.13-2.78(m,5H),2.68-1.89(m,4H),1.42(dd,J＝104.6,66.9Hz,7H),1.11-0.17(m,7H).
实施例20
(S)-5-((R)-3-环丙基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(恶唑-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.32,tR＝0.36min.
1H NMR(400MHz,DMSO)δ10.77(s,1H),7.95(s,1H),7.19(s,1H),4.64(s,1H),3.19(dd,J＝12.2,6.2Hz,2H),2.38(dd,J＝12.9,8.5Hz,1H),2.01(dd,J＝12.8,6.7Hz,1H),0.67(d,J＝9.3Hz,4H).
13C NMR(101MHz,DMSO)δ167.30,152.62,136.22,125.77,59.63,51.56,37.35,20.08,10.36,9.71.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率22％。
LC-MS(ESI):[M+1]+＝379.52,tR＝1.83min.
1H NMR(400MHz,CDCl3)δ7.57-7.39(m,1H),7.02(d,J＝15.1Hz,1H),5.32(d,J＝17.8Hz,1H),3.77(d,J＝9.5Hz,2H),3.48(d,J＝9.6Hz,1H),3.12-2.89(m,2H),2.83-2.63(m,2H),1.75-1.13(m,11H),0.96-0.60(m,4H).
实施例21
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己酰基)-N-(5-甲基异恶唑-3基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝221.79,tR＝0.93min.
1H NMR(400MHz,DMSO)δ10.71(s,1H),6.62(s,1H),4.56(s,1H),3.17(d,J＝11.9Hz,2H),2.39(s,2H),2.33(dd,J＝12.8,8.5Hz,1H),1.98(dd,J＝12.9,7.0Hz,1H),0.65(d,J＝14.3Hz,4H).
13C NMR(101MHz,DMSO)δ170.05,167.12,157.36,96.21,59.47,51.57,37.49,20.16,12.10,10.16,9.88.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率32％。
LC-MS(ESI):[M+1]+＝393.22,tR＝1.69min.
1H NMR(400MHz,DMSO)δ11.00(d,J＝65.0Hz,1H),8.45-7.49(m,1H),6.61(s,1H),4.66(d,J＝79.9Hz,1H),3.73-3.15(m,5H),2.43(d,J＝56.1Hz,5H),2.20-1.69(m,2H),1.66-1.02(m,6H),0.86(s,3H),0.58(d,J＝30.4Hz,4H).
实施例22
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己羰基)-N-(噻唑-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.38,tR＝0.92min.
1H NMR(400MHz,DMSO)δ10.84(s,1H),7.53(d,J＝3.1Hz,1H),7.34(d,J＝3.2Hz,1H),4.79-4.59(m,1H),3.21(ddd,J＝15.7,10.7,5.6Hz,2H),2.37(dd,J＝12.9,8.4Hz,1H),2.03(dd,J＝13.0,6.9Hz,1H),0.77-0.57(m,4H).
13C NMR(101MHz,DMSO)δ167.07,157.46,137.44,114.41,59.19,51.59,37.46,20.16,10.22,9.94.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率26％。
LC-MS(ESI):[M+1]+＝395.04，tR＝1.69min.
1H NMR(400MHz,CDCl3)δ7.88(s,1H),7.83(s,1H),7.49-6.90(m,1H),5.38(d,J＝29.3Hz,1H),4.10(dd,J＝25.4,13.2Hz,1H),4.01(d,J＝8.4Hz,1H),3.41(dd,J＝33.5,11.2Hz,2H),3.14(s,1H),2.49-2.25(m,1H),1.76(d,J＝5.9Hz,1H),1.39-1.26(m,6H),1.02-0.80(m,3H),0.69-0.53(m,4H).
实施例23
(S)-5-((R)-2-((N-羟甲酰胺基)甲基)己酰基)-N-(1,3,4-噻二唑-2-基)-5-氮杂螺[2.4]庚烷‐6‐甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.87,tR＝0.63min.
1H NMR(400MHz,DMSO)δ10.85(dd,J＝52.8,47.6Hz,1H),9.13(t,J＝28.8Hz,1H),4.96-4.50(m,1H),3.47-2.92(m,2H),2.46-1.89(m,2H),1.15-0.23(m,4H).
13C NMR(101MHz,DMSO)δ167.60,158.24,149.57,59.31,51.59,37.35,20.10,10.31,9.77.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率32％。
LC-MS(ESI):[M+1]+＝396.18,tR＝1.53min.
1H NMR(400MHz,CDCl3)δ8.53(s,1H),7.57(s,1H),5.39-4.77(s,1H),4.76-4.21(m,1H),4.20-3.63(m,1H),3.61-3.44(m,2H),3.43-3.28(m,2H),3.27-2.74(m,1H),2.70-2.31(m,1H),1.52-1.17(m,2H),1.12-0.88(m,4H),0.87-0.54(m,3H),0.52-0.03(m,4H).
实施例24
N-((R)-2-((S)-6-(1-(2-(N,N-二甲基)乙基)-1H-苯并[d]咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-羰基)己酰基)-N-羟基甲酰胺的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
步骤4：取步骤3中产物加入THF溶解，然后加入2.5倍当量的NaH，30分钟后加入1.5倍当量二甲氨基氯乙烷，50℃下回流反应，反应完毕，加入水，EA，萃取，无水硫酸钠干燥后浓缩过柱得产物。
LC-MS(ESI):[M+1]+＝285.17,tR＝0.57min.
1H NMR(400MHz,D2O)δ7.80-7.61(m,2H),7.58-7.39(m,2H),5.50(dd,J＝9.1,7.8Hz,1H),4.93-4.76(m,2H),3.69-3.22(m,4H),2.96(s,6H),2.67-2.22(m,2H),0.88-0.63(m,4H).
13C NMR(101MHz,D2O)δ147.40,135.78,132.90,126.31,126.04,117.18,111.46,54.06,52.87,52.72,43.42,39.33,38.01,20.39,11.70,7.58.
步骤5：操作如合成通式(X2)中的步骤3。
步骤6:操作如合成通式(X2)中的步骤4，制备冻干得白色固体，两步收率27％。
LC-MS(ESI):[M+1]+＝456.35,tR＝1.56min.
1H NMR(400MHz,CDCl3)δ8.35(s,1H),7.90-7.72(m,2H),7.40-7.31(m,2H),5.89-5.44(m,1H),5.45-5.21(m,1H),4.67-4.47(m,1H),4.31-3.97(m,3H),3.85-3.72(m,1H),3.55-3.50(m 1H),3.46-3.30(m,1H),2.92-2.66(m,8H),2.16-1.89(m,1H),1.87-1.62(m,1H),1.46-1.34(m,2H),1.32-1.19(m,5H),0.84-0.71(m,3H),0.69-049(m,4H).
实施例25
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(4-吗啉苯基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝302.05,tR＝1.06min.
1H NMR(400MHz,DMSO)δ10.82(s,1H),7.52(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),4.55(t,J＝7.9Hz,1H),3.78-3.67(m,4H),3.19(dd,J＝37.9,11.2Hz,2H),3.08-3.02(m,4H),2.33(dd,J＝12.9,8.2Hz,1H),2.01(dd,J＝12.9,7.6Hz,1H),0.78-0.53(m,4H).
13C NMR(101MHz,DMSO)δ165.87,147.72,130.32,120.50,115.35,66.04,59.59,51.66,48.71,37.81,20.35,10.34,9.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率35％。
LC-MS(ESI):[M+1]+＝499.31,tR＝1.76min.
1H NMR(400MHz,CDCl3)δ7.84(d,J＝17.4Hz,1H),7.54-7.32(m,2H),6.87(dd,J＝13.7,10.3Hz,2H),5.05-4.71(m,1H),4.22-3.80(m,8H),3.23-3.00(m,5H),2.06(d,J＝8.2Hz,4H),1.90-1.37(m,10H),0.75-0.40(m,3H).
实施例26
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙羰基)-N-(吡嗪-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.57min.
1H NMR(400MHz,D2O)δ8.98(d,J＝96.9Hz,1H),8.31(t,J＝81.8Hz,2H),4.66-4.49(m,1H),3.26-2.81(m,2H),2.29-1.62(m,2H),0.63-0.09(m,4H).
13C NMR(101MHz,D2O)δ148.55,144.82,137.12,133.80,60.43,52.76,37.11,20.13,9.77,8.95,8.64.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率32％。
LC-MS(ESI):[M+1]+＝416.21，tR＝1.65min.
1H NMR(400MHz,CDCl3)δ9.48(s,1H),8.42-8.25(m,1H),8.22(d,J＝10.1Hz,1H),7.76(d,J＝46.4Hz,1H),4.93(s,1H),4.03-3.50(m,3H),3.36(m,1H),3.19(d,J＝6.3Hz,1H),2.23(d,J＝12.1Hz,1H),2.02-1.19(m,12H),0.83-0.59(m,4H).
实施例27
(S)-5-((R)-3-环戊基‐2‐((N‐羟甲酰胺基)甲基)丙酰基)‐N‐(嘧啶‐4‐基)‐5‐螺[2.4]庚烷‐6‐甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.43min.
1H NMR(400MHz,D2O)δ9.08(s,1H),8.78(d,J＝6.8Hz,1H),8.42(t,J＝31.2Hz,1H),4.82(dd,J＝8.7,6.4Hz,1H),3.50-3.15(m,2H),2.57-2.00(m,2H),0.88-0.58(m,4H).
13C NMR(101MHz,D2O)δ169.77,152.74,150.17,111.15,60.98,52.82,36.76,20.08,9.91,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率28％。
LC-MS(ESI):[M+1]+＝416.21，tR＝1.26min.
1H NMR(400MHz,CDCl3)δ11.25-9.16(m,2H),9.12-8.46(m,2H),8.42-7.54(m,2H),5.30(s,1H),4.15-3.60(m,2H),3.23(ddt,J＝86.3,30.4,19.1Hz,3H),2.36-1.19(m,13H),0.80-0.24(m,4H).
实施例28
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(5-氟吡啶-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝236.09,tR＝1.00min.
1H NMR(400MHz,D2O)δ8.25(s,1H),7.98-7.87(m,1H),7.64(dd,J＝9.2,3.9Hz,1H),4.75-4.73(m,1H),3.48-3.21(m,2H),2.44(dd,J＝13.3,9.0Hz,1H),2.13(dd,J＝13.3,6.4Hz,1H),0.79-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.10,156.72(d,1JC-F＝250.3Hz),144.96,131.48(d,2JC-F＝30.9Hz),131.10(d,2JC-F＝18.7Hz),118.14(d,3JC-F＝6.2Hz).
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率36％。
LC-MS(ESI):[M+1]+＝434.22,tR＝1.95min.
1H NMR(400MHz,CDCl3)δ10.33(s,1H),8.64-7.75(m,2H),7.61-6.95(m,1H),4.71(s,1H),4.11-3.74(m,1H),3.71-3.49(m,1H),3.46-2.87(m,2H),2.86-2.50(m,1H),2.47-2.07(m,1H),1.72(m,6H),1.53-1.17(m,4H),1.16-0.76(m,2H),0.73-0.21(m,3H).
实施例29
(S)-5-((R)-3-环戊甲基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(5-甲基噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝238.45,tR＝1.09min.
1H NMR(400MHz,D2O)δ7.24(s,1H),4.77(dd,J＝8.9,6.5Hz,1H),3.24(s,2H),2.38(dd,J＝13.4,9.1Hz,1H),2.28(s,3H),2.05(dd,J＝13.4,6.3Hz,1H),0.72-0.45(m,4H).
13C NMR(101MHz,D2O)δ168.01,159.21,129.86,124.06,60.16,52.75,36.68,20.02,11.03,9.78,8.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率35％。
LC-MS(ESI):[M+1]+＝435.24,tR＝1.93min.
1H NMR(400MHz,CDCl3)δ13.75(d,J＝200.5Hz,1H),10.54(s,1H),7.67(dd,J＝86.8,36.9Hz,1H),7.15(d,J＝89.5Hz,1H),5.67-4.98(m,1H),4.85-2.78(m,5H),2.58-1.06(m,17H),0.96-0.31(m,4H).
实施例30
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(3-氟吡啶-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝235.90,tR＝0.60min.
1H NMR(400MHz,D2O)δ8.20(d,J＝5.5Hz,1H),8.06(t,J＝9.0Hz,1H),7.56-7.50(m,1H),4.90-4.79(m,1H),3.31(s,2H),2.61-2.39(m,1H),2.15(dd,J＝13.4,6.1Hz,1H),0.76-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.94,150.12(d,J＝255.5Hz),138.18,137.92(d,J＝13.4Hz),130.12(d,J＝16.9Hz),123.10(d,J＝5.8Hz),60.39,52.83,37.21,20.06,9.96,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率38％。
LC-MS(ESI):[M+1]+＝433.22,tR＝1.65min.
1H NMR(400MHz,CDCl3)δ9.40(d,J＝352.7Hz,1H),8.24(dd,J＝55.9,22.9Hz,1H),7.88(d,J＝28.7Hz,1H),7.65-7.41(m,1H),7.39-7.03(m,1H),5.24-4.45(m,1H),4.24-3.99(m,1H),3.97-3.58(m,2H),2.88(dddd,J＝100.0,91.1,60.9,42.5Hz,2H),2.44-1.37(m,11H),1.18-0.98(m,2H),0.93-0.50(m,4H).
实施例31
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(1H-吡唑-3-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1
步骤2：操作如合成通式(X1)中的步骤2。
1H NMR(400MHz,DMSO)δ11.53(s,1H),10.66(d,J＝4.0Hz,1H),8.94(s,1H),7.79(s,1H),6.52(s,1H),4.70-4.51(m,1H),3.31-3.24(m,1H),3.21-3.15(m,1H),2.43-2.27(m,1H),2.01(dd,J＝12.7,7.5Hz,1H),0.80-0.57(m,4H).
13C NMR(101MHz,DMSO)δ165.97,144.77,96.15,55.94,51.54,37.61,18.42,10.22,9.83.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率31％。
LC-MS(ESI):[M+1]+＝404.56，tR＝1.16min.
1H NMR(400MHz,CDCl3)δ11.40(s,1H),11.18(s,1H),7.92-7.62(m,1H),7.58-7.36(m,1H),6.92(d,J＝66.8Hz,1H),5.51-5.21(m,1H),4.60(t,J＝8.8Hz,1H),4.36-3.96(m,2H),3.81-3.66(m,2H),2.81-2.44(m,2H),2.36-1.62(m,11H),0.76-0.19(m,4H).
实施例32
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-环丙基-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝182.98,tR＝0.550min
1H NMR(400MHz,CDCl3)δ7.82(s,1H),3.93(dd,J＝8.7,5.2Hz,1H),3.00-2.95(m,1H),2.89(s,1H),2.77-2.71(m,2H),2.24-1.78(m,2H),0.97-0.25(m,8H).
13C NMR(101MHz,CDCl3)δ175.92,60.94,54.70,39.27,22.37,22.09,11.00,9.48,6.30,6.27.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率27％。
LC-MS(ESI):[M+1]+＝378.16,tR＝1.60min
1H NMR(400MHz,CDCl3)δ8.67-7.74(m,1H),7.27(s,1H),7.05-6.31(m,1H),4.89-4.33(dt,J＝14.5,10.4Hz,1H),4.04-3.79(m,1H),3.79-3.64(m,1H),3.61-3.13(m,2H),3.13-2.84(m,1H),2.84-2.57(m,1H),2.29-2.05(m,1H),1.88-1.54(m,6H),1.53-1.04(m,4H),1.03-0.18(m,7H).
实施例33
(S)-5-((R)-3-环戊甲基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(4-甲基噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝237.60,tR＝0.66min.
1H NMR(400MHz,DMSO)δ9.09(d,J＝4.8Hz,1H),6.89(d,J＝0.9Hz,1H),4.92-4.46(m,1H),3.42-2.98(m,2H),2.48-1.85(m,5H),0.88-0.41(m,4H).
13C NMR(101MHz,DMSO)δ167.14,157.01,146.07,108.59,59.22,51.52,37.47,20.13,16.53,10.22,9.90.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率29％。
LC-MS(ESI):[M+1]+＝435.24,tR＝1.91min.
1H NMR(400MHz,CDCl3)δ7.87(s,1H),7.27(s,1H),6.75-6.30(m,1H),4.93(s,1H),4.26-3.35(m,6H),2.22-1.36(m,16H),0.83-0.48(m,4H).
实施例34
3-((S)-5-((R)-3‐环戊基‐2‐((N‐羟胺基)甲基)丙酰基)‐5‐螺[2.4]庚烷‐6‐甲酰)噻吩‐2‐羧酸甲酯的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝281.76,tR＝1.85min.
1H NMR(400MHz,DMSO)δ11.86(s,1H),8.11(d,J＝5.4Hz,1H),7.89(d,J＝5.4Hz,1H),3.92(dt,J＝16.2,8.1Hz,1H),3.84(s,3H),2.98(d,J＝10.0Hz,1H),2.71(d,J＝
10.0Hz,1H),2.11(dd,J＝12.4,9.0Hz,1H),1.80(dd,J＝12.4,4.4Hz,1H),0.63-0.33(m,4H).
13C NMR(101MHz,DMSO)δ173.46,163.00,143.39,133.02,121.38,109.96,61.15,54.16,51.94,39.11,22.40,11.14,9.04.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率37％。
LC-MS(ESI):[M+1]+＝478.45,tR＝2.19min.
1H NMR(400MHz,CDCl3)δ11.29-10.18(m,1H),8.61-7.01(m,3H),4.73(ddd,J＝34.4,10.2,5.4Hz,1H),4.17-3.27(m,6H),2.14-1.00(m,14H),0.94-0.37(m,4H).
实施例35
N-((R)-2-(环戊甲基)-3-((S)-6-(1,1-二氧化硫代吗啉-4-羰基)-5-氮杂螺[2.4]庚烷-5-基)-3-丙羰基)-N-羟甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝259.00,tR＝0.37min.
1H NMR(400MHz,DMSO)δ9.94(t,J＝76.9Hz,1H),8.16(t,J＝56.7Hz,1H),4.42-3.90(m,1H),3.64-3.25(m,2H),3.21-2.53(m,8H),2.53-2.01(m,2H),2.01-0.66(m,4H).
13C NMR(101MHz,DMSO)δ167.37,57.55,51.79,50.64,50.55,43.26,40.80,36.27,33.89,20.33,10.13,9.41.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率27％。
LC-MS(ESI):[M+1]+＝456.16，tR＝1.63min.
1H NMR(400MHz,CDCl3)δ7.88(s,1H),4.99(s,1H),4.66(d,J＝13.7Hz,1H),4.26(d,J＝13.2Hz,1H),3.98(s,2H),3.93-3.66(m,3H),3.61-2.94(m,7H),2.09-1.94(m,2H),1.92-1.20(m,10H),0.80-0.50(m,4H).
实施例36
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙羰基)-N-(恶唑-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.32,tR＝0.36min.
1H NMR(400MHz,DMSO)δ10.77(s,1H),7.95(s,1H),7.19(s,1H),4.64(s,1H),3.19(dd,J＝12.2,6.2Hz,2H),2.38(dd,J＝12.9,8.5Hz,1H),2.01(dd,J＝12.8,6.7Hz,1H),0.67(d,J＝9.3Hz,4H).
13C NMR(101MHz,DMSO)δ167.30,152.62,136.22,125.77,59.63,51.56,37.35,20.08,10.36,9.71.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率24％。
LC-MS(ESI):[M+1]+＝405.06，tR＝1.58min.
1H NMR(400MHz,CDCl3)δ8.72(s,1H),7.59(s,1H),7.14(s,1H),5.36(s,1H),4.18(s,1H),3.65(s,1H),3.42-2.94(m,3H),2.20(s,1H),1.93-0.72(m,12H),0.42-0.13(m,4H).
实施例37
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(异恶唑-5-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.11,tR＝0.57min.
1H NMR(400MHz,DMSO)δ10.66(s,1H),9.01(s,1H),8.51(d,J＝1.6Hz,1H),6.29(d,J＝1.6Hz,1H),4.98-4.32(m,1H),3.29-2.98(m,2H),2.45-1.87(m,2H),0.85-0.47(m,4H).
13C NMR(101MHz,DMSO)δ165.75,160.13,152.15,88.25,59.47,51.64,37.35,20.12,10.27,9.78
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率32％。
LC-MS(ESI):[M+1]+＝405.53，tR＝1.32min.
1H NMR(400MHz,CDCl3)δ10.59(d,J＝42.7Hz,1H),8.26-8.04(m,1H),7.81(s,1H),6.44-6.16(m,1H),4.98-4.70(m,1H),3.94-3.69(m,2H),3.56-3.28(m,2H),2.31-2.06(m,2H),1.85-1.08(m,11H),0.84-0.54(m,4H).
实施例38
(S)-N-(5-(叔丁基)异恶唑-3-基)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)‐5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝265.07,tR＝1.44min.
1H NMR(400MHz,D2O)δ6.14(s,1H),4.44-4.41(m,1H),3.12-2.93(m,2H),2.19(dd,J＝13.4,8.9Hz,1H),1.86(dd,J＝13.4,6.1Hz,1H),0.52-0.29(m,4H).
13C NMR(101MHz,D2O)δ183.22,167.90,157.06,93.24,60.23,52.67,37.13,32.51,27.67,20.09,9.86,8.49.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率31％。
LC-MS(ESI):[M+1]+＝461.66，tR＝1.46min.
1H NMR(400MHz,CDCl3)δ11.25(s,1H),9.77(s,1H),8.36(s,1H),8.02-7.46(m,1H),6.68(d,J＝28.5Hz,1H),4.92-4.53(m,1H),4.09-3.73(m,2H),3.63-3.41(m,2H),3.31-2.71(m,2H),1.76-1.25(m,20H),0.80-0.49(m,4H).
实施例39
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙羰基)-N-(5-甲基异恶唑-3-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝221.79,tR＝0.93min.
1H NMR(400MHz,DMSO)δ10.71(s,1H),6.62(s,1H),4.56(s,1H),3.17(d,J＝11.9Hz,2H),2.39(s,2H),2.33(dd,J＝12.8,8.5Hz,1H),1.98(dd,J＝12.9,7.0Hz,1H),0.65(d,J＝14.3Hz,4H).
13C NMR(101MHz,DMSO)δ170.05,167.12,157.36,96.21,59.47,51.57,37.49,20.16,12.10,10.16,9.88.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率16％。
LC-MS(ESI):[M+1]+＝419.11，tR＝1.84min.
1H NMR(400MHz,CDCl3)δ7.81(s,1H),7.18(s,1H),4.73(d,J＝74.6Hz,1H),3.73(d,J＝67.7Hz,2H),3.25(m,3H),2.38(s,3H),2.05(m,2H),1.66-1.16(m,11H),0.57(m,4H).
实施例40
(S)-5-((S)-3-环戊基-2-((N-羟甲酰胺)甲基)丙酰基)-N-(噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.38,tR＝0.92min.
1H NMR(400MHz,DMSO)δ10.84(s,1H),7.53(d,J＝3.1Hz,1H),7.34(d,J＝3.2Hz,1H),4.79-4.59(m,1H),3.21(ddd,J＝15.7,10.7,5.6Hz,2H),2.37(dd,J＝12.9,8.4Hz,1H),2.03(dd,J＝13.0,6.9Hz,1H),0.77-0.57(m,4H).
13C NMR(101MHz,DMSO)δ167.07,157.46,137.44,114.41,59.19,51.59,37.46,20.16,10.22,9.94.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，色谱制备(DCM:MeOH＝10:1)得白色固体，两步收率42％。
LC-MS(ESI):[M+1]+＝421.00,tR＝1.84min.
1H NMR(400MHz,CDCl3)δ11.84(s,1H),10.46(s,1H),8.51-7.68(m,1H),7.34(d,J＝60.6Hz,1H),7.04-6.56(m,1H),5.26-4.62(m,1H),4.43-2.80(m,5H),2.34-1.68(m,6H),1.44-0.98(m,7H),0.72(ddt,J＝59.3,52.3,23.7Hz,4H).
实施例41
(S)-5-((R)-3-环戊基-2-((N-羟甲酰胺基)甲基)丙酰基)-N-(1,3,4-噻二唑-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.87,tR＝0.63min.
1H NMR(400MHz,DMSO)δ10.85(dd,J＝52.8,47.6Hz,1H),9.13(t,J＝28.8Hz,1H),4.96-4.50(m,1H),3.47-2.92(m,2H),2.46-1.89(m,2H),1.15-0.23(m,4H).
13C NMR(101MHz,DMSO)δ167.60,158.24,149.57,59.31,51.59,37.35,20.10,10.31,9.77.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率33％。
LC-MS(ESI):[M+1]+＝422.14,tR＝1.65min.
1H NMR(400MHz,CDCl3)δ8.80(s,1H),7.27(s,1H),6.06-5.12(m,1H),5.15-4.62(m,1H),4.29-3.82(m,1H),3.79-3.02(m,3H),2.99-2.34(m,1H),2.31-2.15(m,1H),2.10-1.50(m,7H),1.48-1.26(m,5H),1.17-0.95(m,2H),0.86-0.69(m,2H).
实施例42
N-((R)-3-((S)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-基)-2-(环戊甲基)-3-丙羰基)-N-羟甲酰胺基的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
LC-MS(ESI):[M+1]+＝214.16,tR＝0.94min.
1H NMR(400MHz,D2O)δ7.67-7.58(m,2H),7.49-7.39(m,2H),5.41(t,J＝8.6Hz,1H),3.55-3.46(m,1H),3.25-3.15(m,1H),2.68-2.57(m,1H),2.39-2.28(m,1H),0.85-0.60(m,4H).
13C NMR(101MHz,D2O)δ144.90,130.95,127.05,114.21,36.52,20.20,11.92,7.25.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4，柱层析(DCM:MeOH＝10:1)得白色固体，两步收率33％。
LC-MS(ESI):[M+1]+＝412.18，tR＝1.60min.
1H NMR(400MHz,CDCl3)δ8.44-7.11(m,5H),5.58(s,1H),4.04(s,1H),3.89-2.64(m,4H),2.51-2.02(m,2H),1.95-0.94(m,11H),0.89-0.29(m,4H).
实施例43
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(4-吗啉苯基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝302.05,tR＝1.06min.
1H NMR(400MHz,DMSO)δ10.82(s,1H),7.52(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),4.55(t,J＝7.9Hz,1H),3.78-3.67(m,4H),3.19(dd,J＝37.9,11.2Hz,2H),3.08-3.02(m,4H),2.33(dd,J＝12.9,8.2Hz,1H),2.01(dd,J＝12.9,7.6Hz,1H),0.78-0.53(m,4H).
13C NMR(101MHz,DMSO)δ165.87,147.72,130.32,120.50,115.35,66.04,59.59,51.66,48.71,37.81,20.35,10.34,9.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝514.83,tR＝1.56min.
1H NMR(400MHz,CDCl3)δ7.34(s,1H),6.90(s,1H),5.06(s,1H),4.46(s,1H),3.71(d,J＝18.0Hz,3H),3.17(d,J＝17.6Hz,3H),2.82(d,J＝28.7Hz,1H),2.36(s,1H),1.97(s,1H),1.67-1.08(m,3H),0.89-0.52(m,4H).
实施例44
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(吡嗪-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.57min.
1H NMR(400MHz,D2O)δ8.98(d,J＝96.9Hz,1H),8.31(t,J＝81.8Hz,2H),4.66-4.49(m,1H),3.26-2.81(m,2H),2.29-1.62(m,2H),0.63-0.09(m,4H).
13C NMR(101MHz,D2O)δ148.55,144.82,137.12,133.80,60.43,52.76,37.11,20.13,9.77,8.95,8.64.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝390.52,tR＝1.36min.
1H NMR(400MHz,DMSO)δ12.86-11.39(m,1H),10.31(d,J＝98.3Hz,1H),7.83-6.53(m,3H),5.52-4.73(m,1H),4.11(dd,J＝44.5,40.2Hz,1H),3.94-3.35(m,2H),3.03-0.74(m,13H),0.71-0.24(m,4H).
实施例45
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(嘧啶-4-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.43min.
1H NMR(400MHz,D2O)δ9.08(s,1H),8.78(d,J＝6.8Hz,1H),8.42(t,J＝31.2Hz,1H),4.82(dd,J＝8.7,6.4Hz,1H),3.50-3.15(m,2H),2.57-2.00(m,2H),0.88-0.58(m,4H).
13C NMR(101MHz,D2O)δ169.77,152.74,150.17,111.15,60.98,52.82,36.76,20.08,9.91,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝390.52,tR＝1.38min.
1H NMR(400MHz,CDCl3)δ8.97(s,1H),8.78(s,1H),7.67(s,1H),4.94(s,1H),3.52-2.98(m,3H),2.89-2.34(m,3H),1.92-1.09(m,7H),0.89(s,3H),0.68-0.42(m,4H).
实施例46
(S)-N-(5-氟吡啶-2-基)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝236.09,tR＝1.00min.
1H NMR(400MHz,D2O)δ8.25(s,1H),8.11-7.80(m,1H),7.65(dd,J＝9.1,3.7Hz,1H),4.74(d,J＝7.9Hz,1H),3.44-3.15(m,2H),2.59-1.94(m,2H),0.84-0.47(m,4H).
13C NMR(101MHz,D2O)δ169.09,157.99,155.50,144.99,118.16,118.10,60.33,52.77,37.08,20.12,9.73,8.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率36％。
LC-MS(ESI):[M+1]+＝407.52,tR＝1.93min.
1H NMR(400MHz,CDCl3)δ10.41(d,J＝191.4Hz,1H),9.42(d,J＝136.3Hz,1H),8.11(dt,J＝70.7,24.8Hz,2H),7.65-7.17(m,1H),5.20-4.40(m,1H),4.16-1.83(m,7H),1.79-0.97(m,9H),0.97-0.21(m,6H).
实施例47
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(5-甲基噻唑-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝238.45,tR＝1.09min.
1H NMR(400MHz,D2O)δ7.24(s,1H),4.77(dd,J＝8.9,6.5Hz,1H),3.24(s,2H),2.38(dd,J＝13.4,9.1Hz,1H),2.28(s,3H),2.05(dd,J＝13.4,6.3Hz,1H),0.72-0.45(m,4H).
13C NMR(101MHz,D2O)δ168.01,159.21,129.86,124.06,60.16,52.75,36.68,20.02,11.03,9.78,8.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率25％。
LC-MS(ESI):[M+1]+＝409.53,tR＝1.38min.
1H NMR(400MHz,DMSO)δ12.02(d,J＝79.3Hz,1H),10.37(s,1H),9.07-8.35(m,1H),7.38-6.92(m,1H),5.51-4.35(m,1H),3.83-3.38(m,2H),3.34(s,2H),2.97-2.61(m,1H),2.54-2.43(m,2H),2.40-2.28(m,2H),2.28-1.67(m,4H),1.56-1.06(m,6H),0.99-0.81(m,2H),0.78-0.19(m,4H).
实施例48
(S)-5-((R)-2-(2-(羟氨基)-2-氧代乙基)己酰基)-N-(哒嗪-3-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.55min.
1H NMR(400MHz,D2O)δ9.19(d,J＝5.0Hz,1H),8.78(d,J＝9.2Hz,1H),8.29(dd,J＝9.2,5.2Hz,1H),4.92-4.78(m,1H),3.47-3.15(m,2H),2.50(dd,J＝13.4,9.1Hz,1H),2.20(dd,J＝13.4,6.3Hz,1H),0.85-0.59(m,4H).
13C NMR(101MHz,D2O)δ169.33,155.57,145.59,134.68,128.12,60.68,52.82,36.96,20.13,9.84,8.65.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝390.26,tR＝1.68min.
1H NMR(400MHz,CDCl3)δ10.42(s,1H),8.88(s,1H),8.44(t,J＝36.8Hz,1H),7.50(s,1H),5.17-4.79(m,1H),3.83-3.65(m,1H),3.58(dd,J＝19.5,9.4Hz,1H),3.18(d,J＝7.0Hz,1H),2.41-2.16(m,2H),2.04(t,J＝18.5Hz,1H),1.62(d,J＝4.2Hz,1H),1.50-1.36(m,6H),0.84(s,3H),0.75-0.48(m,4H).
实施例49
(S)-N-(3-氟吡啶-2-基)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝235.90,tR＝0.60min.
1H NMR(400MHz,D2O)δ8.20(d,J＝5.5Hz,1H),8.06(t,J＝9.0Hz,1H),7.56-7.50(m,1H),4.90-4.79(m,1H),3.31(s,2H),2.61-2.39(m,1H),2.15(dd,J＝13.4,6.1Hz,1H),0.76-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.94,150.12(d,J＝255.5Hz),138.18,137.92(d,J＝13.4Hz),130.12(d,J＝16.9Hz),123.10(d,J＝5.8Hz),60.39,52.83,37.21,20.06,9.96,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率22％。
LC-MS(ESI):[M+1]+＝407.33,tR＝1.63min.
1H NMR(400MHz,DMSO)δ10.57-10.32(m,1H),10.30-10.06(m,1H),9.11-8.61(m,1H),8.59-7.77(m,2H),7.82-7.13(m,1H),5.25-4.52(m,1H),3.75-3.52(m,2H),3.34(s,2H),3.22-3.01(m,2H),2.95-2.57(m,1H),2.36-1.70(m,2H),1.55-1.25(m,7H),1.11-0.35(m,4H).
实施例50
(S)-N-环丙基-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝182.98,tR＝0.550min
1H NMR(400MHz,CDCl3)δ7.82(s,1H),3.93(dd,J＝8.7,5.2Hz,1H),3.00-2.95(m,1H),2.89(s,1H),2.77-2.71(m,2H),2.24-1.78(m,2H),0.97-0.25(m,8H).
13C NMR(101MHz,CDCl3)δ175.92,60.94,54.70,39.27,22.37,22.09,11.00,9.48,
6.30,6.27.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率25％。
LC-MS(ESI):[M+1]+＝352.33,tR＝1.47min
1H NMR(400MHz,CDCl3)δ6.13-5.33(m,1H),3.72(d,J＝6.1Hz,2H),3.33-2.98(m,2H),2.64-1.84(m,1H),1.70-1.62(m,1H),1.52-1.41(m,12H),1.39-1.21(m,4H),1.19-0.52(m,3H).
实施例51
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(4-甲基噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.43min.
1H NMR(400MHz,DMSO)δ9.09(d,J＝4.8Hz,1H),6.89(d,J＝0.9Hz,1H),4.92-4.46(m,1H),3.42-2.98(m,2H),2.48-1.85(m,5H),0.88-0.41(m,4H).
13C NMR(101MHz,DMSO)δ167.14,157.01,146.07,108.59,59.22,51.52,37.47,20.13,16.53,10.22,9.90.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率29％。
LC-MS(ESI):[M+1]+＝409.34,tR＝1.84min.
1H NMR(400MHz,DMSO)δ12.02(d,J＝79.3Hz,1H),10.37(s,1H),9.07-8.35(m,1H),7.38-6.92(m,1H),5.51-4.35(m,1H),3.83-3.38(m,2H),3.34(s,2H),2.97-2.61(m,
1H),2.54-2.43(m,2H),2.40-2.28(m,2H),2.28-1.67(m,4H),1.56-1.06(m,6H),0.99-0.81(m,2H),0.78-0.19(m,4H).
实施例52
(S)-N-(1-(2-(二甲氨基)乙基)-1H-吡唑-4-基)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝278.38,tR＝1.10min.
1H NMR(400MHz,D2O)δ8.01-7.76(m,1H),7.59(d,J＝22.1Hz,1H),4.53-4.38(m,2H),3.69-3.39(m,3H),2.86-2.73(m,6H),2.41-2.27(m,1H),2.11-1.94(m,1H),1.32-0.90(m,2H),0.74-0.53(m,4H).
13C NMR(101MHz,D2O)δ167.17,133.04,123.88,119.97,56.50,52.60,48.83,46.22,43.03,37.19,20.12,9.59,8.76.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝449.35,tR＝1.13min.
1H NMR(500MHz,CDCl3)δ7.86(s,1H),7.18(s,1H),5.30(d,J＝161.0Hz,3H),3.75(d,J＝12.0Hz,3H),3.28-2.38(m,10H),2.27-0.72(m,11H),0.37(d,J＝91.8Hz,4H).
实施例53
(S)-5-((R)-2-(2-(羟氨基)-2-氧代乙基)己酰基)-N-(恶唑-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.32,tR＝0.36min.
1H NMR(400MHz,DMSO)δ10.77(s,1H),7.95(s,1H),7.19(s,1H),4.64(s,1H),3.19(dd,J＝12.2,6.2Hz,2H),2.38(dd,J＝12.9,8.5Hz,1H),2.01(dd,J＝12.8,6.7Hz,1H),0.67(d,J＝9.3Hz,4H).
13C NMR(101MHz,DMSO)δ167.30,152.62,136.22,125.77,59.63,51.56,37.35,20.08,10.36,9.71.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率23％。
LC-MS(ESI):[M+1]+＝379.23,tR＝1.49min.
1H NMR(400MHz,DMSO)δ11.57-10.73(m,1H),10.59-9.67(m,1H),9.10-8.40(m,1H),7.86(d,J＝7.9Hz,1H),7.17-6.58(m,1H),3.71-3.47(m,1H),3.34(s,1H),3.03-2.77(m,1H),2.70(d,J＝8.0Hz,4H),2.55-2.41(m,2H),2.29-1.79(m,2H),1.63-0.89(m,7H),0.85(t,J＝6.5Hz,2H),0.63-0.14(m,2H).
实施例54
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(异恶唑-5-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.11,tR＝0.57min.
1H NMR(400MHz,DMSO)δ10.66(s,1H),9.01(s,1H),8.51(d,J＝1.6Hz,1H),6.29(d,J＝1.6Hz,1H),4.98-4.32(m,1H),3.29-2.98(m,2H),2.45-1.87(m,2H),0.85-0.47(m,4H).
13C NMR(101MHz,DMSO)δ165.75,160.13,152.15,88.25,59.47,51.64,37.35,20.12,10.27,9.78
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝379.23,tR＝1.53min.
1H NMR(400MHz,CDCl3)δ10.99(t,J＝150.4Hz,1H),9.89(s,1H),8.45-7.95(m,1H),7.27(s,1H),6.63-6.16(m,1H),5.05-4.48(m,1H),3.98-3.33(m,2H),3.11-1.04(m,12H),1.01-0.28(m,6H).
实施例55
(S)-N-(5-(叔丁基)异恶唑-3-基)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝265.07,tR＝1.44min.
1H NMR(400MHz,D2O)δ6.14(s,1H),4.44-4.41(m,1H),3.12-2.93(m,2H),2.19(dd,J＝13.4,8.9Hz,1H),1.86(dd,J＝13.4,6.1Hz,1H),0.52-0.29(m,4H).
13C NMR(101MHz,D2O)δ183.22,167.90,157.06,93.24,60.23,52.67,37.13,32.51,27.67,20.09,9.86,8.49.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率23％。
LC-MS(ESI):[M+1]+＝435.30,tR＝2.02min.
1H NMR(400MHz,CDCl3)δ11.01(d,J＝134.6Hz,1H),10.47-9.87(m,1H),6.74-6.29(m,1H),5.65(s,1H),5.05-4.42(m,1H),3.96-3.16(m,2H),3.05-2.71(m,2H),2.51-1.95(m,2H),1.36-1.13(m,18H),0.90-0.50(m,4H).
实施例56
(S)-5-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(5-甲基异恶唑-3-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝221.79,tR＝0.93min.
1H NMR(400MHz,DMSO)δ10.71(s,1H),6.62(s,1H),4.56(s,1H),3.17(d,J＝11.9Hz,2H),2.39(s,2H),2.33(dd,J＝12.8,8.5Hz,1H),1.98(dd,J＝12.9,7.0Hz,1H),0.65(d,J＝14.3Hz,4H).
13C NMR(101MHz,DMSO)δ170.05,167.12,157.36,96.21,59.47,51.57,37.49,20.16,12.10,10.16,9.88.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率29％。
LC-MS(ESI):[M+1]+＝393.42,tR＝1.79min.
1H NMR(500MHz,CDCl3)δ6.59(s,1H),4.98(s,1H),4.06(d,J＝161.2Hz,2H),3.46-2.23(m,7H),1.96(s,1H),1.72-1.14(m,6H),1.03-0.28(m,7H).
实施例57
(S)-5-((R)-2‐(2‐(羟胺基)‐2‐氧代乙基)己酰基)‐N‐(5‐(三氟甲基)噻唑‐2‐基)‐5‐螺[2.4]庚烷‐6‐甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝292.22,tR＝1.28min
1H NMR(400MHz,DMSO)δ10.46(s,1H),8.09(d,J＝0.9Hz,1H),4.67(dd,J＝8.1,7.0Hz,1H),3.26-3.17(m,2H),2.36(dd,J＝13.2,8.5Hz,1H),2.06(dd,J＝13.2,6.6Hz,1H),0.76-0.61(m,4H).
13C NMR(101MHz,DMSO)δ167.88,159.26,121.96,119.23,117.58,59.25,51.77,37.12,20.13,10.32,9.60.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率32％。
LC-MS(ESI):[M+1]+＝463.08,tR＝2.00min.
1H NMR(400MHz,CDCl3)δ12.92-11.05(m,1H),10.14(s,1H),8.07-6.63(m,1H),5.60-4.50(m,1H),3.71(ddd,J＝49.1,46.6,9.5Hz,2H),3.46-2.27(m,2H),2.08(ddd,J＝54.9,36.3,19.1Hz,2H),1.80-1.12(m,6H),1.10-0.31(m,7H).
实施例58
(R)-3-((S)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-甲酰基)-N-羟庚酰胺的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
LC-MS(ESI):[M+1]+＝214.16,tR＝0.94min.
1H NMR(400MHz,D2O)δ7.67-7.58(m,2H),7.49-7.39(m,2H),5.41(t,J＝8.6Hz,1H),3.55-3.46(m,1H),3.25-3.15(m,1H),2.68-2.57(m,1H),2.39-2.28(m,1H),0.85-0.60(m,4H).
13C NMR(101MHz,D2O)δ144.90,130.95,127.05,114.21,36.52,20.20,11.92,7.25.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。
步骤6:操作如合成通式(X2)中的步骤5，三步收率17％。
LC-MS(ESI):[M+1]+＝385.49,tR＝1.65min.
1H NMR(400MHz,DMSO)δ7.48(s,2H),7.15(t,J＝21.2Hz,2H),5.48-5.07(m,1H),3.88-3.53(m,2H),3.41(dd,J＝23.6,16.5Hz,2H),2.81(d,J＝51.3Hz,1H),2.40-2.11(m,2H),1.84-1.00(m,9H),0.79-0.30(m,4H).
实施例59
(R)-3-((R)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-羰基)-N-羟庚酰胺基的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
LC-MS(ESI):[M+1]+＝385.49,tR＝1.65min.
1H NMR(400MHz,CDCl3)δ7.57(s,2H),7.22(s,2H),5.43(dd,J＝36.1,29.6Hz,1H),3.93-3.79(m,1H),3.48-3.31(m,1H),3.09-2.28(m,5H),2.07-1.90(m,1H),1.24(dd,J＝26.4,11.2Hz,5H),0.78-0.61(m,7H).
实施例60
(R)-3-((S)-6-(1-(2-(二甲氨基)乙基)-1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-羰基)-N-羟庚基酰胺的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
LC-MS(ESI):[M+1]+＝285.17,tR＝0.57min.
1H NMR(400MHz,D2O)δ7.80-7.61(m,2H),7.58-7.39(m,2H),5.50(dd,J＝9.1,7.8Hz,1H),4.93-4.76(m,2H),3.69-3.22(m,4H),2.96(s,6H),2.67-2.22(m,2H),0.88-0.63(m,4H).
13C NMR(101MHz,D2O)δ147.40,135.78,132.90,126.31,126.04,117.18,111.46,54.06,52.87,52.72,43.42,39.33,38.01,20.39,11.70,7.58.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。
步骤6:操作如合成通式(X2)中的步骤5，三步收率17％。
LC-MS(ESI):[M+1]+＝456.34tR＝1.23min.
1H NMR(400MHz,DMSO)δ10.34(d,J＝47.3Hz,1H),9.91(d,J＝51.5Hz,1H),8.00-7.62(m,2H),7.57-7.26(m,2H),5.40(dd,J＝26.3,18.9Hz,1H),5.03-4.68(m,2H),4.02(d,J＝9.4Hz,1H),3.76-3.66(m,1H),3.64-3.50(m,2H),3.04-2.76(m,6H),2.50(s,2H),2.48-1.85(m,4H),1.46-1.17(m,7H),1.20-0.99(m,2H),0.86-0.58(m,6H).
实施例61
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(4-吗啉苯基)-5-氮杂螺[2.4]庚烷-6-
甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝302.05,tR＝1.06min.
1H NMR(400MHz,DMSO)δ10.82(s,1H),7.52(d,J＝8.8Hz,2H),6.93(d,J＝8.8Hz,2H),4.55(t,J＝7.9Hz,1H),3.78-3.67(m,4H),3.19(dd,J＝37.9,11.2Hz,2H),3.08-3.02(m,4H),2.33(dd,J＝12.9,8.2Hz,1H),2.01(dd,J＝12.9,7.6Hz,1H),0.78-0.53(m,4H).
13C NMR(101MHz,DMSO)δ165.87,147.72,130.32,120.50,115.35,66.04,59.59,51.66,48.71,37.81,20.35,10.34,9.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率23％。
LC-MS(ESI):[M+1]+＝499.68,tR＝1.68min.
1H NMR(400MHz,DMSO)δ10.38(s,1H),9.96-9.56(m,1H),8.70(s,1H),7.43(t,J＝8.7Hz,2H),6.81(t,J＝44.5Hz,2H),4.63-4.33(m,1H),3.72(s,2H),3.60(dd,J＝31.0,9.4Hz,1H),3.22(d,J＝11.5Hz,2H),3.03(d,J＝3.8Hz,2H),2.50(s,8H),2.25-0.45(m,15H).
实施例62
(S)-5-((R)-2-(环戊甲基)-4-(羟胺)-4-氧代丁基)-N-(吡嗪-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.57min.
1H NMR(400MHz,D2O)δ8.98(d,J＝96.9Hz,1H),8.31(t,J＝81.8Hz,2H),4.66-4.49(m,1H),3.26-2.81(m,2H),2.29-1.62(m,2H),0.63-0.09(m,4H).
13C NMR(101MHz,D2O)δ148.55,144.82,137.12,133.80,60.43,52.76,37.11,20.13,9.77,8.95,8.64.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率27％。
LC-MS(ESI):[M+1]+＝416.59,tR＝1.59min.
1H NMR(400MHz,CDCl3)δ10.15(s,1H),8.46-7.88(m,3H),4.78(s,1H),3.88(dd,J＝28.6,9.4Hz,1H),3.64-3.39(m,1H),3.24-2.54(m,3H),2.08-1.91(m,1H),1.87-1.17(m,12H),0.81-0.46(m,4H).
实施例63
(S)-5-((R)-2-(环戊甲基)-4-(羟氨基)-4-氧代丁基)-N-(嘧啶-4-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.43min.
1H NMR(400MHz,D2O)δ9.08(s,1H),8.78(d,J＝6.8Hz,1H),8.42(t,J＝31.2Hz,1H),4.82(dd,J＝8.7,6.4Hz,1H),3.50-3.15(m,2H),2.57-2.00(m,2H),0.88-0.58(m,4H).
13C NMR(101MHz,D2O)δ169.77,152.74,150.17,111.15,60.98,52.82,36.76,20.08,9.91,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率24％。
LC-MS(ESI):[M+1]+＝416.57,tR＝1.54min.
1H NMR(400MHz,CDCl3)δ10.08(s,1H),8.85(s,1H),8.58(t,J＝5.0Hz,1H),8.13(s,1H),4.83(s,1H),3.84(d,J＝9.8Hz,1H),3.73-2.64(m,4H),2.13(d,J＝5.8Hz,1H),1.96-1.20(m,12H),0.72-0.45(m,4H).
实施例64
(S)-5-((R)-2-(环戊甲基)-4-(羟氨基)-4-氧代丁基)-N-(5-氟吡啶-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝236.09,tR＝1.00min.
1H NMR(400MHz,D2O)δ8.25(s,1H),7.98-7.87(m,1H),7.64(dd,J＝9.2,3.9Hz,1H),4.75-4.73(m,1H),3.48-3.21(m,2H),2.44(dd,J＝13.3,9.0Hz,1H),2.13(dd,J＝13.3,6.4Hz,1H),0.79-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.10,156.72(d,1JC-F＝250.3Hz),144.96,131.48(d,2JC-F＝30.9Hz),131.10(d,2JC-F＝18.7Hz),118.14(d,3JC-F＝6.2Hz).
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率18％。
LC-MS(ESI):[M+1]+＝433.56,tR＝1.56min.
1H NMR(400MHz,CDCl3)δ9.60(s,1H),8.36-7.72(m,2H),7.43-7.12(m,1H),4.82(d,J＝63.6Hz,1H),3.88(dd,J＝25.2,9.4Hz,1H),3.27-2.40(m,4H),2.01-1.86(m,1H),1.85-1.18(m,12H),0.72-0.50(m,4H).
实施例65
(S)-5-((R)-2-(环戊甲基)-4-(羟氨基)-4-氧代丁基)-N-(5-甲基噻唑-2-基)-5-氮杂螺[2.4]庚烷-6-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝238.45,tR＝1.09min.
1H NMR(400MHz,D2O)δ7.24(s,1H),4.77(dd,J＝8.9,6.5Hz,1H),3.24(s,2H),2.38(dd,J＝13.4,9.1Hz,1H),2.28(s,3H),2.05(dd,J＝13.4,6.3Hz,1H),0.72-0.45(m,4H).
13C NMR(101MHz,D2O)δ168.01,159.21,129.86,124.06,60.16,52.75,36.68,20.02,11.03,9.78,8.73.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率19％。
LC-MS(ESI):[M+1]+＝435.59,tR＝1.35min.
1H NMR(400MHz,CDCl3)δ10.34(s,1H),7.00(s,1H),5.01-4.58(m,1H),3.95-3.64
(m,1H),3.54(d,J＝10.0Hz,1H),3.10-2.82(m,1H),2.58-2.11(m,5H),1.87-0.91(m,12H),0.72-0.58(m,4H).
实施例66
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(哒嗪-3-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝219.14,tR＝0.55min.
1H NMR(400MHz,D2O)δ9.19(d,J＝5.0Hz,1H),8.78(d,J＝9.2Hz,1H),8.29(dd,J＝9.2,5.2Hz,1H),4.92-4.78(m,1H),3.47-3.15(m,2H),2.50(dd,J＝13.4,9.1Hz,1H),2.20(dd,J＝13.4,6.3Hz,1H),0.85-0.59(m,4H).
13C NMR(101MHz,D2O)δ169.33,155.57,145.59,134.68,128.12,60.68,52.82,36.96,20.13,9.84,8.65.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝416.59,tR＝1.48min.
1H NMR(400MHz,CDCl3)δ11.83-9.51(m,2H),9.12-8.70(m,1H),8.46(dd,J＝30.4,8.8Hz,1H),7.69-7.34(m,1H),5.44-4.71(m,1H),4.31-0.87(m,18H),0.83-0.17(m,4H).
实施例67
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(3-氟吡啶-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝235.90,tR＝0.60min.
1H NMR(400MHz,D2O)δ8.20(d,J＝5.5Hz,1H),8.06(t,J＝9.0Hz,1H),7.56-7.50(m,1H),4.90-4.79(m,1H),3.31(s,2H),2.61-2.39(m,1H),2.15(dd,J＝13.4,6.1Hz,1H),0.76-0.54(m,4H).
13C NMR(101MHz,D2O)δ169.94,150.12(d,J＝255.5Hz),138.18,137.92(d,J＝13.4Hz),130.12(d,J＝16.9Hz),123.10(d,J＝5.8Hz),60.39,52.83,37.21,20.06,9.96,8.50.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝433.56,tR＝1.49min.
1H NMR(400MHz,CDCl3)δ10.25(d,J＝63.9Hz,1H),9.75(s,1H),8.23(dd,J＝59.2,44.9Hz,1H),7.45(tq,J＝16.9,8.5Hz,1H),7.09(dd,J＝26.9,23.5Hz,1H),4.82(d,J＝52.3Hz,1H),4.04-3.29(m,2H),3.02-0.93(m,16H),0.92-0.31(m,4H).
实施例68
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-环丙基-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝182.98,tR＝0.550min
1H NMR(400MHz,CDCl3)δ7.82(s,1H),3.93(dd,J＝8.7,5.2Hz,1H),3.00-2.95(m,1H),2.89(s,1H),2.77-2.71(m,2H),2.24-1.78(m,2H),0.97-0.25(m,8H).
13C NMR(101MHz,CDCl3)δ175.92,60.94,54.70,39.27,22.37,22.09,11.00,9.48,6.30,6.27.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率25％。
LC-MS(ESI):[M+1]+＝378.59,tR＝1.49min.
1H NMR(400MHz,DMSO)δ10.37(d,J＝8.2Hz,1H),8.88-8.46(m,1H),7.89-7.36(m,1H),4.55-4.09(m,1H),3.69-3.37(m,2H),3.38-3.26(m,2H),3.25-2.89(m,1H),2.87-2.55(m,2H),2.42-2.12(m,1H),2.08-1.81(m,2H),1.80-1.24(m,9H),1.00-0.24(m,8H).
实施例69
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(恶唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.32,tR＝0.36min.
1H NMR(400MHz,DMSO)δ10.77(s,1H),7.95(s,1H),7.19(s,1H),4.64(s,1H),3.19(dd,J＝12.2,6.2Hz,2H),2.38(dd,J＝12.9,8.5Hz,1H),2.01(dd,J＝12.8,6.7Hz,1H),0.67(d,J＝9.3Hz,4H).
13C NMR(101MHz,DMSO)δ167.30,152.62,136.22,125.77,59.63,51.56,37.35,20.08,10.36,9.71.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率25％。
LC-MS(ESI):[M+1]+＝405.49,tR＝1.48min.
1H NMR(400MHz,CDCl3)δ7.60(s,1H),7.15(s,1H),5.07(s,1H),3.63-3.00(m,3H),2.89-2.39(m,3H),1.93-1.19(m,12H),0.62-0.48(m,4H).
实施例70
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(异恶唑-5-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝208.32,tR＝0.36min.
1H NMR(400MHz,DMSO)δ10.77(s,1H),7.95(s,1H),7.19(s,1H),4.64(s,1H),3.19(dd,J＝12.2,6.2Hz,2H),2.38(dd,J＝12.9,8.5Hz,1H),2.01(dd,J＝12.8,6.7Hz,1H),0.67(d,J＝9.3Hz,4H).
13C NMR(101MHz,DMSO)δ167.30,152.62,136.22,125.77,59.63,51.56,37.35,20.08,10.36,9.71.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝405.46,tR＝1.53min.
1H NMR(400MHz,CDCl3)δ11.40-9.93(m,1H),8.54-7.96(m,1H),6.56-6.17(m,1H),4.92-4.22(m,1H),4.11-3.12(m,2H),3.10-0.92(m,15H),0.91-0.22(m,4H).
实施例71
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(5-甲基异恶唑-3-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝221.79,tR＝0.93min.
1H NMR(400MHz,DMSO)δ10.71(s,1H),6.62(s,1H),4.56(s,1H),3.17(d,J＝11.9Hz,2H),2.39(s,2H),2.33(dd,J＝12.8,8.5Hz,1H),1.98(dd,J＝12.9,7.0Hz,1H),0.65(d,J＝14.3Hz,4H).
13C NMR(101MHz,DMSO)δ170.05,167.12,157.36,96.21,59.47,51.57,37.49,20.16,12.10,10.16,9.88.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率45％。
LC-MS(ESI):[M+1]+＝419.59,tR＝1.32min.
1H NMR(400MHz,CDCl3)δ10.80-9.61(m,2H),7.27(s,1H),6.83-6.31(m,1H),5.32(d,J＝16.2Hz,1H),4.76(d,J＝35.4Hz,1H),4.32-1.82(m,12H),1.79-1.10(m,6H),1.06-0.20(m,6H).
实施例72
(S)-5-((R)-2-(环戊甲基)-4-(羟氨基)-4-氧代丁酰基)-N-(噻唑-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.38,tR＝0.92min.
1H NMR(400MHz,DMSO)δ10.84(s,1H),7.53(d,J＝3.1Hz,1H),7.34(d,J＝3.2Hz,1H),4.79-4.59(m,1H),3.21(ddd,J＝15.7,10.7,5.6Hz,2H),2.37(dd,J＝12.9,8.4Hz,1H),2.03(dd,J＝13.0,6.9Hz,1H),0.77-0.57(m,4H).
13C NMR(101MHz,DMSO)δ167.07,157.46,137.44,114.41,59.19,51.59,37.46,20.16,10.22,9.94.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率21％。
LC-MS(ESI):[M+1]+＝421.46,tR＝1.46min.
1H NMR(400MHz,DMSO)δ12.13(s,1H),10.35(d,J＝21.1Hz,1H),8.87-8.56(m,1H),7.46(d,J＝3.2Hz,1H),7.20(d,J＝3.2Hz,1H),4.65(t,J＝7.0Hz,1H),3.80-3.41(m,2H),2.99-2.75(m,1H),2.23(dd,J＝14.7,8.7Hz,1H),2.11-1.95(m,2H),1.92-1.63(m,4H),1.50(ddd,J＝27.3,13.9,6.3Hz,5H),1.21(dd,J＝13.7,7.3Hz,2H),1.04(ddd,J＝19.6,11.1,7.4Hz,2H),0.76-0.59(m,3H),0.58-0.45(m,2H).
实施例73
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(5-(三氟甲基)噻唑-2-基)-5-螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝292.22,tR＝1.28min
1H NMR(400MHz,DMSO)δ10.46(s,1H),8.09(d,J＝0.9Hz,1H),4.67(dd,J＝8.1,7.0Hz,1H),3.26-3.17(m,2H),2.36(dd,J＝13.2,8.5Hz,1H),2.06(dd,J＝13.2,6.6Hz,1H),0.76-0.61(m,4H).
13C NMR(101MHz,DMSO)δ167.88,159.26,121.96,119.23,117.58,59.25,51.77,37.12,20.13,10.32,9.60.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝489.17,tR＝2.12min.
1H NMR(400MHz,CDCl3)δ13.14-11.09(m,1H),10.14(d,J＝58.3Hz,1H),7.44(dd,J＝111.4,81.1Hz,2H),5.29-4.55(m,1H),4.17-3.27(m,2H),3.24-2.51(m,2H),2.48-0.84(m,14H),0.82-0.28(m,3H).
实施例74
(S)-5-((R)-2-(环戊甲基)-4-(羟胺基)-4-氧代丁基)-N-(1,3,4-噻二唑-2-基)-5-氮杂螺[2.4]庚烷-6-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝224.87,tR＝0.63min.
1H NMR(400MHz,DMSO)δ10.85(dd,J＝52.8,47.6Hz,1H),9.13(t,J＝28.8Hz,1H),4.96-4.50(m,1H),3.47-2.92(m,2H),2.46-1.89(m,2H),1.15-0.23(m,4H).
13C NMR(101MHz,DMSO)δ167.60,158.24,149.57,59.31,51.59,37.35,20.10,10.31,9.77.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率23％。
LC-MS(ESI):[M+1]+＝422.14,tR＝1.58min.
1H NMR(400MHz,DMSO)δ12.64(s,1H),10.35(d,J＝21.9Hz,1H),9.36-9.02(m,1H),8.96-8.41(m,1H),4.89-4.51(m,1H),2.25-1.98(m,2H),1.95-0.95(m,12H),0.94-0.24(m,4H).
实施例75
(R)-4-((S)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-基)-3-(环戊甲基)-N-羟基-4-氧代丁酰胺的合成
步骤1：操作如实施例1中步骤1。
步骤2：操作如实施例1中步骤2。
步骤3：操作如实施例1中步骤3。
LC-MS(ESI):[M+1]+＝214.16,tR＝0.94min.
1H NMR(400MHz,D2O)δ7.67-7.58(m,2H),7.49-7.39(m,2H),5.41(t,J＝8.6Hz,1H),3.55-3.46(m,1H),3.25-3.15(m,1H),2.68-2.57(m,1H),2.39-2.28(m,1H),0.85-0.60(m,4H).
13C NMR(101MHz,D2O)δ144.90,130.95,127.05,114.21,36.52,20.20,11.92,7.25.
步骤4：操作如合成通式(X2)中的步骤3。
步骤5:操作如合成通式(X2)中的步骤4。
步骤6:操作如合成通式(X2)中的步骤5，三步收率16％。
LC-MS(ESI):[M+1]+＝411.53,tR＝1.34min.
1H NMR(400MHz,CDCl3)δ7.97-7.47(m,2H),7.49-7.28(m,1H),7.21(dd,J＝15.3,12.4Hz,2H),5.66-5.29(m,1H),3.93-3.26(m,2H),3.18-2.90(m,2H),2.87-2.58(m,1H),2.55-2.10(m,2H),1.94-0.99(m,11H),0.81-0.25(m,4H).
实施例76
(R)-4-((R)-6-(1H-苯并咪唑-2-基)-5-氮杂螺[2.4]庚烷-5-基)-3-(环戊甲基)-N-羟基-4-氧代丁酰胺的合成
制备分离所得旋转异构体
LC-MS(ESI):[M+1]+＝411.53,tR＝1.32min.
1H NMR(400MHz,CDCl3)δ12.46(s,1H),7.53(s,2H),7.16(s,2H),5.25(s,1H),3.84(s,1H),2.98(d,J＝42.9Hz,2H),2.74-2.31(m,3H),2.08(s,1H),1.88-1.06(m,11H),0.69-0.52(m,4H).
实施例77
(S)-2-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(吡嗪-2-基)-2-氮杂螺[4.4]壬烷-3-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝246.92,tR＝1.02min.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率23％。
LC-MS(ESI):[M+1]+＝418.5,tR＝1.38min.
1H NMR(400MHz,DMSO)δ11.17-10.66(m,1H),10.24(d,J＝95.0Hz,1H),9.30(s,1H),8.38(d,J＝17.7Hz,2H),4.69-4.37(m,1H),3.95-3.06(m,2H),3.02-2.64(m,1H),2.42-1.76(m,4H),1.74-0.43(m,17H).
实施例78
(S)-N-(5-氟吡啶-2-基)-2-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-2-螺[4.4]壬烷-3-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝263.93,tR＝1.28min.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率20％。
LC-MS(ESI):[M+1]+＝435.18,tR＝2.03min.
1H NMR(400MHz,CDCl3)δ9.77(d,J＝529.3Hz,2H),7.82-6.38(m,2H),4.73(s,1H),3.96-2.84(m,3H),2.63-1.91(m,6H),1.89-0.45(m,17H).
实施例79
(S)-2-((R)-2-(2-(羟氨基)-2-氧代乙基)己酰基)-N-(5-甲基噻唑-2-基)-2-氮杂螺[4.4]壬烷-3-酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝268.09,tR＝1.21min.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率21％。
LC-MS(ESI):[M+1]+＝437.39,tR＝2.02min.
1H NMR(400MHz,CDCl3)δ7.18(s,1H),5.25(s,1H),3.36-3.28(m,2H),3.13-3.08(m,1H),2.63(dd,J＝24.7,10.3Hz,1H),2.51-2.31(m,4H),1.87-1.13(m,16H),0.89(s,3H).
实施例80
(S)-2-((R)-2-(2-(羟胺基)-2-氧代乙基)己酰基)-N-(5-甲基异恶唑-3-基)-2-螺[4.4]壬烷-3-甲酰胺的合成
步骤1：操作如合成通式(X1)中的步骤1。
步骤2：操作如合成通式(X1)中的步骤2。
LC-MS(ESI):[M+1]+＝249.23,tR＝0.98min.
1H NMR(400MHz,DMSO)δ10.71(s,1H),6.62(s,1H),4.56(s,1H),3.17(d,J＝11.9Hz,2H),2.39(s,2H),2.33(dd,J＝12.8,8.5Hz,1H),1.98(dd,J＝12.9,7.0Hz,1H),0.65(d,J＝14.3Hz,4H).
13C NMR(101MHz,DMSO)δ170.05,167.12,157.36,96.21,59.47,51.57,37.49,20.16,
12.10,10.16,9.88.
步骤3：操作如合成通式(X2)中的步骤3。
步骤4:操作如合成通式(X2)中的步骤4。
步骤5:操作如合成通式(X2)中的步骤5，三步收率30％。
LC-MS(ESI):[M+1]+＝421.31,tR＝1.49min.
1H NMR(400MHz,DMSO)δ10.88(s,1H),6.62(d,J＝31.4Hz,1H),4.48-4.05(m,1H),3.33(s,8H),2.50(dt,J＝3.5,1.7Hz,9H),1.87-0.99(m,8H),0.96-0.73(m,2H).
对照实验一
B、C化合物来源于专利CN101584694A和CN101869563A。
表8.三元环引入对活性的影响
Sybyl打分是利用Sybyl X2软件的打分函数给出的，分数越高，说明预测的最低抑菌浓度越低。结合实验所测得的最低抑菌浓度，不难发现三元环的引入对活性提高的作用是明显的。
对照实验二
由于LBM415、GSK1322322等PDF抑制剂在代谢过程中会产生芳香羟胺、芳香肼类活性物种，进而造成人体高铁血红蛋白症的产生。为此本专利采用了连续波长法方法，扫描了血样在紫外可见光分光光度计500至700nm波长范围内的吸收谱，然后计算三个健康志愿者血样经所合成的化合物作用后高铁血红蛋白的含量。
实验材料：烟酰胺腺嘌呤二核苷酸磷酸(NADPH)还原体系，包括葡萄糖-6-磷酸二钠、烟酰胺腺嘌呤二核苷磷酸和葡萄糖-6-磷酸脱氢酶(购自上海源叶生物技
术公司)加到1mL磷酸缓冲溶液(PBS,pH 7.4)中于37℃摇床型水浴中摇晃10min，冷至0℃。混合人员肝细胞组分S9购自Xenotech公司(20mg S9蛋白/1mL悬浮介质)。化合物用配成50mM浓度的储备液，-20℃保存待用。Dapsone用作阳性对照实验试剂，购自SigmaAldrich。1％w/v triton X-100自配。血样用含EDTA抗凝剂的Vacutainer管收集(BD,Franklin Lakes,美国)，来自三个健康志愿者，采集后立即使用。
血样与药物孵育体系配制：250μL孵育体系由220μL全血、25μL S9(终浓度：2mg S9蛋白/mL孵化液),2.5μL NADPH形成体系(终浓度：10mM G-6-P,1mM NADP,7.5units/mL)和2.5μL化合物储备液(终浓度500μM)组成。取2mL孵育体系放于Eppendorf管中，于37℃水浴下摇晃孵化5小时。
高铁血红蛋白的含量测定：取60μL孵化后的样品，用3.0mL 1％Triton X-100裂解后，用紫外-可见光分光光度计测定高铁血红蛋白(MetHb)的含量。Triton X-100溶液做空白对照。样品放置于比色皿后1分钟，记录待测样光谱，波长从500nm到700nm；加入铁***2mg后，静止2分钟，使样品血中亚铁完全氧化后，铁***饱和的高铁血红蛋白的标准谱。高铁血红蛋白形成百分含量(MetHb％)通过630nm处吸光度值比来获得。MetHb％＝(A'-AB')/AM'其中，A'为630nm处样品的总吸光度，AB'为630nm处样品中还原型血的基值吸光度，AM'为630nm处铁***饱和的氧化型血的吸光度。
表9.化合物体外高铁血红蛋白毒性的评价
| 实施例化合物 | MetHb％(n＝3) |
| 全血采样后立即测定 | 1.00±0.02 |
| 经孵育但不加药物的全血 | 1.10±0.07 |
| 实施例2 | 3.32±0.05 |
| 实施例3 | 3.33±0.06 |
| 实施例6 | 2.25±0.02 |
| 实施例7 | 2.24±0.05 |
| 实施例8 | 4.12±0.07 |
| 实施例9 | 1.13±0.06 |
| 实施例10 | 1.12±0.01 |
| 实施例11 | 1.26±0.07 |
| 实施例12 | 1.22±0.02 |
| 实施例13 | 1.15±0.06 |
| LBM415 | 3.05±0.01 |
| GSK1322322 | 3.00±0.02 |
体外毒性实验结果表明，与对照药LBM415、GSK1322322相比，合成的螺三元环化合物2、3、6、7、8和9对血液的高铁血红蛋白形成概率较低；酰胺生物电子等排体唑类化合物的毒性非常小。
对照实验三
表10.实施例化合物和对照化合物对革兰氏阴性菌卡他莫拉菌的MIC对比
相较于对照化合物，所合成的化合物对革兰氏阴性菌卡他莫拉菌的抗菌活性有很明显地提高。
Claims (18)
- 一种螺三元环、螺五元环类肽脱甲酰基酶抑制剂，其特征在于，所述抑制剂结构如式(1)所示：
式(1)中，n＝2-4，R1为正丁基、环戊甲基；R2为芳香环，芳香杂环，杂环，烷基；R3为氢或烷基。 - 如权利要求1所述的抑制剂，其特征在于，式(1)中，n＝2和4；R1为正丁基、环戊甲基；R2为1H‐吡唑‐3‐基、5‐氟吡啶1‐氧化物‐2‐基、5‐(叔丁基)异恶唑‐3‐基、6‐甲基‐N‐(4‐(吡啶‐3‐基)嘧啶‐2‐基)苯‐1‐氨基‐3‐基、3‐氟吡啶‐2‐基、5‐甲基噻唑‐2‐基、3‐(吡啶‐3‐基)苯基、N‐(嘧啶‐2‐基)苯‐1‐氨基‐3‐基、4‐(吡啶‐3‐基)嘧啶‐2‐基、4‐***啉苯基、2‐吡嗪基、3‐哒嗪基、4‐嘧啶基、1‐甲基‐1H‐吡唑‐4‐基、5‐异恶唑基、环丙基、4‐甲基噻唑‐2‐基、2‐恶唑基、5‐甲基异恶唑‐3‐基、2‐噻唑基、1,3,4‐硫二唑‐2‐基、5‐(三氟甲基)噻唑‐2‐基、双乙基砜基、苯并噻唑‐2‐基、3‐甲酸甲酯‐2‐噻吩基；R3为氢。
- 一种螺三元环、螺五元环类肽脱甲酰基酶抑制剂，其特征在于，所述抑制剂结构如式(2)所示：
式(2)中，n＝2-4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。 - 如权利要求3所述的抑制剂，其特征在于，式(2)中，n＝2和4；R1为正丁基、环戊甲基；R2为2-苯并咪唑基、1,3,4-氧杂二恶唑、1,2,4-氧杂二恶唑、1,3,4-三氮唑。
- 一种螺三元环、螺五元环类肽脱甲酰基酶抑制剂，其特征在于，所述抑制剂结构如式(3)所示：
式(3)中，n＝2‐4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环，杂环，烷基；R3为氢或烷基。 - 如权利要求5所述的抑制剂，其特征在于，式(3)中，n＝2和4；R1为正丁基、环戊甲基；R2为1H‐吡唑‐3‐基、5‐氟吡啶1‐氧化物‐2‐基、5‐(叔丁基)异恶唑‐3‐基、6‐甲基‐N‐(4‐(吡啶‐3‐基)嘧啶‐2‐基)苯‐1‐氨基‐3‐基、3‐氟吡啶‐2‐基、5‐甲基噻唑‐2‐基、3‐(吡啶‐3‐基)苯基、N‐(嘧啶‐2‐基)苯‐1‐氨基‐3‐基、4‐(吡啶‐3‐基)嘧啶‐2‐基、4‐***啉苯基、2‐吡嗪基、3‐哒嗪基、4‐嘧啶基、1‐甲基‐1H‐吡唑‐4‐基、5‐异恶唑基、环丙基、4‐甲基噻唑‐2‐基、2‐恶唑基、5‐甲基异恶唑‐3‐基、2‐噻唑基、1,3,4‐硫二唑‐2‐基、5‐(三氟甲基)噻唑‐2‐基、双乙基砜基、苯并噻唑‐2‐基、3‐甲酸甲酯‐2‐噻吩基；R3为氢。
- 一种螺三元环、螺五元环类肽脱甲酰基酶抑制剂，其特征在于，所述抑制剂结构如式(4)所示：
式中n＝2‐4，R1为正丁基，环戊甲基；R2为芳香环或芳香杂环。 - 如权利要求7所述的抑制剂，其特征在于，式(4)中，n＝2和4；R1为正丁基、环戊甲基；R2为2‐苯并咪唑基、1,3,4‐氧二恶唑、1,2,4‐氧二恶唑、1,3,4‐三氮唑。
- 一种螺三元环、螺五元环类肽脱甲酰基酶抑制剂，其特征在于，所述抑制剂包括：
- 如权利要求1～9之任一项所述的螺三元环、螺五元环类肽脱甲酰基酶抑制剂在制备抗细菌、抗肿瘤、抗寄生虫的药物中的应用。
- 如权利要求10所述的应用，其特征在于，所述抗细菌是指所述螺三元环、螺五元环类肽脱甲酰基酶抑制剂通过抑制细菌肽脱甲酰基酶来抑制细菌的生长。
- 如权利要求10所述的应用，其特征在于，所述细菌包括金黄色葡萄球菌，表皮葡萄球菌、粪肠球菌、屎肠球菌、肺炎链球菌、卡他莫拉菌。
- 如权利要求10所述的应用，其特征在于，所述抗肿瘤是指所述螺三元环、螺五元环类肽脱甲酰基酶抑制剂用于抑制肿瘤细胞的增殖。
- 如权利要求13所述的应用，其特征在于，所述螺三元环、螺五元环类肽脱甲酰基酶抑制剂用于抑制肿瘤细胞线粒体中的肽脱甲酰基酶，影响肿瘤细胞的能量平衡使得肿瘤细胞线粒体膜去极化，ATP耗竭和促使肿瘤细胞凋亡。
- 如权利要求13所述的应用，其特征在于，所述肿瘤包括结直肠癌、白血病、肺癌、胃癌、***、乳腺癌、***癌和肝癌和骨肉瘤。
- 如权利要求13所述的应用，其特征在于，所述肿瘤细胞包括HCT-116结直肠癌细胞株、Jurkat T细胞白血病细胞株、HL60急性髓系白血病细胞株、A549肺癌细胞株、MGC-803胃癌细胞株、Hela***细胞株、MCF-7乳腺癌细胞株、PC3***癌细胞株、BEL-7402肝癌细胞株和SAOS-2骨肉瘤细胞株。
- 如权利要求10所述的应用，其特征在于，所述抗寄生虫是指所述螺三元环、螺五元环类肽脱甲酰基酶抑制剂通过抑制寄生虫肽脱甲酰基酶来抑制寄生虫的生长。
- 一种药物组合物，其特征在于，其包括如权利要求1～9之任一项所述的螺三元环、螺五元环类肽脱甲酰基酶抑制剂以及药学上可接受的载体。
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| US16/300,191 US10738028B2 (en) | 2016-05-11 | 2017-05-10 | Spiro three-membered ring, spiro five-membered ring peptide deformylase inhibitor and use thereof in antibacteria and anti-tumor |
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Also Published As
| Publication number | Publication date |
|---|---|
| EP3483155A1 (en) | 2019-05-15 |
| ES2930814T3 (es) | 2022-12-22 |
| US20190194165A1 (en) | 2019-06-27 |
| EP3483155A4 (en) | 2020-01-15 |
| EP3483155B1 (en) | 2022-10-26 |
| US10738028B2 (en) | 2020-08-11 |
| JP7001898B2 (ja) | 2022-01-20 |
| JP2019515048A (ja) | 2019-06-06 |
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