Patent Description:
Indoleamine-<NUM>, <NUM>-dioxygenase (IDO) is a monomeric enzyme containing heme discovered by Hayaishi group in <NUM>. The cDNA encoded protein consists of <NUM> amino acids at a molecular weight of <NUM> kDa, which is a rate-limiting enzyme in the catabolism of the tryptophan-kynurenine and widely expressed in many mammalian tissues. In tumor cells, IDO often plays an important role in inducing tumor microenvironment immune tolerance, whose tryptophan (TRP) - kynurenine (KYN) metabolic pathway is involved in the tumor immune escape; IDO also plays an important role in inducing tumor microenvironment immune tolerance.

Tryptophan, as one of the most important essential amino acids in mammals, needs to be taken massively from food to maintain cell activation and proliferation as well as the synthesis of protein and some neurotransmitters, whose deficiency, therefore, can result in the dysfunction of some important cells. IDO can catalyze the conversion of tryptophan to N'-formyl-L-Kynurenine in vivo and degrade the content of tryptophan, which results in the deficiency of tryptophan in vivo and leads to the occurrence of tumors. However, immunohistology suggests that the kynurenine pathway can lead to the increase of quinolinic acid, an excitotoxin as well as many serious human diseases such as Alzheimer.

There are two kinds of tryptophan rate-limiting enzymes in mammals: Tryptophan dioxygenase (TDO) and IDO. In <NUM>, Kotake et al. purified the protein from rabbit intestines and found that TDO is mainly expressed in mammalian liver for the first time. So far, it has not been found yet that TDO is closely correlated with the immune system. TDO can catalyze the kynurenine pathway and convert tryptophan to N'-formyl-L-Kynurenine. In <NUM>, the enzyme purified from rabbit intestines was identified as a dioxygenase (IDO) containing heme. IDO is the only enzyme that can catalyze the oxidative cleavage of indoles in tryptophan molecules and prolong the catabolism of kynurenine pathway in addition to the liver. IDO is usually expressed in organs with more mucous membranes, such as lung, small intestine, large intestine, rectum, spleen, kidney, stomach and brain. In such special/pathological conditions as pregnancy, chronic infection, organ transplantation and tumor, the expression of IDO will be significantly increased, involved in the local immunosuppression.

Studies suggested that IDO can inhibit local T cell immune response in the tumor microenvironment in the following ways: Tryptophan depletion, toxic metabolism and induction of regulatory T cell proliferation. Frequently, it is overexpressed in tumors, consuming local tryptophan and producing a great number of metabolites such as kynurenine. In fact, in the condition of culture without tryptophan or kynurenine, T cell's proliferation will be inhibited, which will decrease in its activity or even end up with apoptosis. There is a very sensitive regulatory point in T cells to tryptophan content. Under the effect of IDO, tryptophan can be consumed, which makes T cells stagnate in the middle of G1 phase, thus inhibiting the T cell proliferation and their immune response. Once T cells stop proliferating, they may not be stimulated again, which is the immune mechanism of IDO in vivo.

<CIT> discloses IDO1 inhibitor compounds of Formula (I) and pharmaceutically acceptable salts thereof, their pharmaceutical compositions, their methods of preparation, and methods for their use in the prevention and/or treatment of diseases.

<CIT> discloses IDO inhibitors and methods for preparing them. The pharmaceutical compositions comprising such IDO inhibitors and methods of using them for treating cancer, infectious diseases, and other disorders are also disclosed.

<CIT> discloses compounds that modulate or inhibit the enzymatic activity of indoleamine <NUM>,<NUM>-dioxygenase (IDO), pharmaceutical compositions containing said compounds and methods of treating proliferative disorders, such as cancer, viral infections and/or inflammatory disorders utilizing the compounds of the disclosure.

<CIT> discloses hetero-aromatic compounds and applications thereof in pharmacy. Specifically, the disclosure provides hetero-aromatic compounds, and steric isomers, geometric isomers, tautomers, racemic bodies, nitrogen oxides, hydrates, solvates, metabolism products, and pharmaceutically acceptable salts or prodrug thereof.

A new type of IDO inhibitor with high activity still waits to be developed in the field, and a novel compound of spiro structure has been found with unexpectedly high IDO inhibitory activity in the invention.

Aspects not falling within the scope of the claims are disclosed and for information only. The invention aims to provide a novel series of compounds of spiro structure as efficient IDO enzyme inhibitors.

Disclosed herein but not covered by the claimed invention is a preparation method of such compounds.

In the first aspect, the present invention provides a compound selected from the group of.

The compound of Formula (I) may be as shown Formula (II):
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as stated in Formula (I); R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, OC<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl respectively; Z represents O or NOH.

The compound of Formula (I) may be as shown Formula (III):
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as described Formula (I); R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl, and C<NUM>-C<NUM> alkynylene aryl.

The compound of Formula (I) may be as shown as Formula (IV):
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as for Formula (I); R<NUM>, R<NUM> and R<NUM> represent hydrogen, C<NUM>-C<NUM> alkyl, OC<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl respectively; R<NUM> and R<NUM> can form <NUM>-<NUM> membered rings or <NUM>-<NUM> membered heterocyclic rings, the heteroatoms in which may be sulfur, oxygen, NH or NRb; n represents an integer of <NUM> to <NUM>; Z represents O or NOH.

The compound of Formula (I) may be as shown as Formula (V):
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as for Formula (I); R<NUM>, R<NUM> and R<NUM> represent hydrogen, C<NUM>-C<NUM> alkyl, OC<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl respectively; R<NUM> and R<NUM> can form <NUM>-<NUM> membered rings or <NUM>-<NUM> membered heterocyclic rings, the heteroatoms in which may be sulfur, oxygen, NH or NRb; n represents an integer to <NUM> to <NUM>; Z represents O or NOH.

The compound of Formula (I) may be shown as Formula (VI),
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as for Formula (I); R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, OC<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl respectively; Z represents O or NOH.

The compound of Formula (I) may be shown as Formula (VII):
<CHM>.

Wherein, R<NUM>, Ring A, Ring B and B are defined as stated in Claim <NUM>; R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, OC<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl respectively; Z represents O or NOH.

The compound of Formula (I) may be shown as Formula (VIII):
<CHM>.

Wherein, R<NUM> represents halogen, and R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene aryl; Z represents O or NOH; Ar<NUM> represents substituted/non-substituted phenyl, and the substituent can be selected from halogen, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> halogenated alkyl, C<NUM>-C<NUM> alkoxy, hydroxyl, amino, nitro, -CF<NUM>, -CN, -SF<NUM>, NRaRb, carboxyl, -CORa, -CO<NUM>C<NUM>-C<NUM> alkyl, -CONRaRb, - SO<NUM>Re, -S02NRaRb, -P(O)Me<NUM>, and -P(O)(Me)<NUM>, wherein, Ra and Rb are defined as for Formula (I).

The compound of Formula (I) may be shown as Formula (IX):
<CHM>.

Wherein, R<NUM> represents halogen, and R<NUM> represents hydrogen, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> cycloalkyl and C<NUM>-C<NUM> alkylene-aryl; Z represents O or NOH; Ar<NUM> represents substituted/non-substituted phenyl, and the substituent can be selected from halogen, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> halogenated alkyl, C<NUM>-C<NUM> alkoxy, hydroxyl, amino, nitro, -CF<NUM>, -CN, -SF<NUM>, NRaRb, carboxyl, -CORa, -CO<NUM>C<NUM>-C<NUM> alkyl, -CONRaRb, - SO<NUM>Re, -S02NRaRb, -P(O)Me<NUM>, and -P(O)(Me)<NUM>, wherein, Ra and Rb are defined as for Formula (I).

In a preferred embodiment, the stereoisomer is a cis-trans isomer.

In another preferred embodiment, the compound is a racemate.

In another preferred embodiment, the stereoisomer is an enantiomer.

It is disclosed herein but not covered by the claimed invention that any hydrogen in the compound may be substituted by deuterium.

In another preferred embodiment, the pharmaceutically acceptable salt is selected from the groups as follows: hydrochloride, hydrobromide, sulfate, phosphate, mesylate, trifluoromethylsulfonate, benzenesulfonate, p-toluenesulfonate (tosylate), <NUM>-Naphthalenesulfonate, <NUM>-naphthalenesulfonate, acetate, trifluoroacetate, malate, tartrate, citrate, lactate, oxalate, succinate, fumarate, maleate, benzoate, salicylate, phenylacetate and mandelate.

The compound in Formula (I) can be obtained with the following preparation methods, including the steps as follows:.

Wherein, R represents C<NUM>-C<NUM> alkyl and C<NUM>-C<NUM> halogenated alkyl; the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other groups or atoms are defined the way above mentioned.

<CHM>
herein, R represents C<NUM>-C<NUM> alkyl and C<NUM>-C<NUM> halogenated alkyl; the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other radical groups or atoms are defined the way above mentioned.

Wherein, the definition of -CHR- is the same as that of V; the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other groups or atoms are defined the way as above mentioned.

Wherein, the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar.

Wherein, the definition of CHR is the same as that of V; the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other groups or atoms are defined the way above mentioned.

Wherein, the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other groups or atoms are defined the way above mentioned.

Wherein, the definition of CHR is the same as that of V; the definition of Ar<NUM> is the same as that of B; the definition of Ar<NUM> is the same as that of Ar, and the other groups or atoms are defined the way as above mentioned.

The base can be selected from the groups as follows: alkali hydroxides, alkali-earth hydroxides, alkali hydride, HMDS alkali metal salt, pyridine, triethylamine, etc..

The acids can be selected from the groups as follows: Hydrochloride, sulfuric acid, etc..

The coupling reagents can be selected from the groups as follows: HATU, etc..

The palladium catalyst can be selected from the groups as follows: tetrakispalladium, etc..

The catalyst can be selected from the groups as follows: palladium-carbon catalyst, etc..

The reductant can be selected from the groups as follows: LiAlH<NUM>, etc..

On the other hand, the invention provides a compound or its stereoisomer/tautomer mentioned in the first aspect or a pharmaceutically acceptable salt, which are used in:.

The invention provides the compounds of the first aspect, or their stereoisomers or tautomers, or pharmaceutically-acceptable salts for use as an antitumor drug. The invention further provides the compounds of the first aspect, or their stereoisomers or tautomers, or pharmaceutically-acceptable salts for use in the prevention and/or treatment of indoleamine-<NUM>,<NUM>-dioxygenase mediated diseases.

In another preferred embodiment, the indoleamine-<NUM>, <NUM>-dioxygenase mediated diseases are those with pathological characteristics of IDO mediated tryptophan metabolism pathway.

The indoleamine-<NUM>, <NUM>-dioxygenase mediated diseases are selected from cancer, neurodegenerative disease, HIV infections, eye disease, psychological disorder, depression, anxiety disorder, Alzheimer's disease and/or autoimmune diseases.

In another preferred embodiment, the cancer includes but not limited to: colon cancer, breast cancer, gastric cancer, lung cancer, colorectal cancer, pancreatic cancer, ovarian cancer, prostate cancer, kidney cancer, liver cancer, brain cancer, melanoma, multiple myeloma, chronic myeloid leukemia, hematologic tumor, lymphoma, including metastases in other tissues or organs far away from the primary tumor lesions.

The invention also provides a pharmaceutical composition comprising:
A compound or its stereoisomer/tautomer mentioned in the first aspect of the invention or a pharmaceutically acceptable salt; and pharmaceutically acceptable carriers.

The pharmaceutical composition also contains other antitumor drugs.

In another preferred embodiment, the other antitumor drugs are selected from the groups as follows: PD-<NUM> antibody, PD-L1 antibody, CTLA-<NUM> antibody and other antitumor chemotherapy drugs and targeted drugs.

In another preferred embodiment, the other antitumor drugs include, but are not limited to, immunotherapeutic drugs against cancer: PD-<NUM> antibody, CTLA-<NUM> antibody, PD-L1 antibody, PD-L2 antibody, any other chemotherapy drug or targeted therapeutic drug, such as HDAC inhibitor, inhibitor of arginine metabolic enzyme, STING activator and EP4 antagonist.

Disclosed herein but not covered by the claimed invention is a prevention and/or treatment of indoleamine-<NUM>, <NUM>-dioxygenase mediated diseases, including the steps giving a patient such a compound of Formula (I) or of the invention as described above or its stereoisomer or tautomer, or its pharmaceutically acceptable salt or prodrug or pharmaceutical composition above.

The indoleamine-<NUM>, <NUM>-dioxygenase mediated diseases may refer to cancers, and the methods further consist of steps of applying additional anticancer agents (also known as antitumor drugs, as described above) to a patient.

The compound of the invention has such pharmacological activities as anti-tumor, treatment of neurodegenerative diseases (Alzheimer's disease), and anti-inflammatory.

It is noteworthy that, within the scope of the invention, the technical features as mentioned above and described in detail below (factual examples) can be combined with each other to form a new or preferred technical solution, which will not be repeated here due to limited length.

With the extensive and in-depth research by the present inventors, a new compound of the structure with spiro features was accidentally developed, which can be used as an efficient IDO enzyme inhibitor to prevent and/or treat indoleamine-<NUM>,<NUM>-dioxygenase mediated diseases, and used as anti-inflammatory drugs. On such basis, the invention was completed.

The term "C<NUM>-C<NUM> alkyl" refers to monovalent saturated aliphatic hydrocarbyl with <NUM>-<NUM> carbon atoms, including straight-chain and branched-chain hydrocarbyl, such as methyl (CH<NUM>-), ethyl (CH<NUM>CH<NUM>-), n-propyl (CH<NUM>CH<NUM>CH<NUM>-), isopropyl ((CH<NUM>)<NUM>CH-), n-butyl (CH<NUM>CH<NUM>CH<NUM>CH<NUM>-), isobutyl ((CH<NUM>)<NUM>CHCH<NUM>-), sec-butyl ((CH<NUM>)(CH<NUM>CH<NUM>)CH-), tert-butyl ((CH<NUM>)<NUM>C-), and n-amyl (CH<NUM>CH<NUM>CH<NUM>CH<NUM>CH<NUM>-), and neopentyl ((CH<NUM>)<NUM>CCH<NUM>-). In the invention, the term includes substituted/non-substituted alkyl.

As used herein, the term " substituted/non-substituted " means that the groups can be non-substituted or that H in the radical group is substituted by one or more (preferably <NUM>-<NUM>, more preferably <NUM>-<NUM>) substituents.

As used herein, the term " substituted/non-substituted " means that the groups have one or more (preferably <NUM>-<NUM>, more preferably <NUM>-<NUM>) substituents selected from the groups as follows: halogen, hydroxyl, -NH<NUM>, nitro, -CN, C<NUM>-C<NUM> alkyl, C<NUM>-C<NUM> halogenated alkyl, C<NUM>-C<NUM> alkoxy, C<NUM>-C<NUM> cycloalkyl, C<NUM>-C<NUM> alkenyl, C<NUM>-C<NUM> alkynyl, phenyl, benzyl, C<NUM>-C<NUM> alkylS(O)<NUM>-, (C<NUM>-C<NUM> alkyl)<NUM>NS(O)<NUM>-, C<NUM>-C<NUM> alkyl-C(O)-, C<NUM>-C<NUM> cycloalkyl-C(O)-, C<NUM>-C<NUM> alkyl-OC(O)-, (C<NUM>-C<NUM> alkyl)<NUM>NC(O)-, C<NUM>-C<NUM> alkyl-C(O)NH-, and (C<NUM>-C<NUM> alkyl)<NUM>NC(O)NH-.

As used herein, the term "C<NUM>-C<NUM> cycloalkyl" refers to a cyclic, substituted/non-substituted cycloalkyl with <NUM>-<NUM> carbon atoms, such as -CH<NUM>-cyclopropane and -CH<NUM>-cyclobutane.

As used herein, the term "alkoxy" refers to -O-alkyl, of which the alkyl may be saturated/unsaturated, may be branched-chain, straight-chain, or cyclic. Preferably, the alkoxy has <NUM>-<NUM> carbon atoms, namely C<NUM>-C<NUM> alkoxys, preferably <NUM>-<NUM> carbon atoms. Representative examples include, but are not limited to: methoxyl, ethoxyl and propoxyl.

As used herein, the term "C<NUM>-C<NUM> aryl" refers to a monocyclic (e.g. phenyl) or condensed ring (e.g. naphthyl or anthracyl) with <NUM>-<NUM> (preferably <NUM>-<NUM>) carbon atoms, and the condensed ring may be nonaromatic (e.g. <NUM>-benzoxazolinone, <NUM>H-<NUM>, <NUM>-benzoxazolinone-<NUM>(<NUM>H)-keone-<NUM>-yl, etc.) if the attachment point is on the aromatic carbon. The preferred aryl consists of phenyl and naphthyl. The term includes substituted/non-substituted forms, of which substituents are defined as above.

As used herein, the term "C<NUM>-C<NUM> alkenyl" refers to an alkenyl with <NUM>-<NUM> (e.g. <NUM>-<NUM> or <NUM>-<NUM>) carbon atoms and at least <NUM> (e.g. <NUM>-<NUM>) unsaturated olefinic bonds (>C = C<). Such radical groups consist of ethenyl, allyl, and but-<NUM>-enyl.

As used herein, the term "C<NUM>-C<NUM> cycloalkyl" refers to a cyclic alkyl with <NUM>-<NUM> carbon atoms and single/multiple rings (including fused, bridged and spiro systems). In a fused ring system, one or more rings can be cycloalkyl, heterocyclic, aryl or heteroaryl, provided the connecting sites are cycloalkyl rings. Examples of suitable cycloalkyls include: Adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclooctyl.

As used herein, the term "halogenated"/"halogen" refers to fluorine, chlorine, bromine and iodine.

As used herein, the term "heteroaryl" refers to aromatic radical groups with <NUM>-<NUM> carbon atoms and <NUM>-<NUM> heteroatoms selected from oxygen, nitrogen and sulfur in the rings. As for the terms of trees indicating carbon atoms, for example, "C<NUM>-C<NUM> heteroaryl" denotes aromatic groups with <NUM>-<NUM> carbon atoms and <NUM>-<NUM> heteroatoms selected from oxygen, nitrogen and sulfur. Others are similar. Such heteroaryl can be monocyclic (e.g., pyridyl or furyl) or condensed ring (such as indolizinyl or benzothiophene), of which the condensed rings can be non aromatic and/or contain a heteroatom, provided the connecting sites are atoms of the aromatic heteroaryl. In one example, ring nitrogen and/or sulfur of heteroaryl are selectively oxidized to N-oxide (N-O), sulfinyl or sulfonyl. Preferably, heteroaryls consists of pyridyl, pyrryl, indolyl, thienyl and furyl. The term includes substituted/non-substituted heteroaryl.

As used herein, the term "substituted heteroaryl" refers to a heteroaryl substituted by <NUM>-<NUM>, preferably <NUM>-<NUM>, more preferably <NUM>-<NUM> substituents selected from the substituent defined similarly with substituted aryl.

As used herein, the term "heterocyclic ring"/"heterocyclic"/"heterocyclic alkyl"/"heterocyclyl" refers to a saturated, partially saturated or unsaturated radical group (but not aromatic), with single rings or condensed rings (including a bridged ring system and a spiro system) in which there are <NUM>-<NUM> carbon atoms and <NUM>-<NUM> (e.g. <NUM>) heteroatoms selected from nitrogen, sulfur or oxygen. In a condensed ring system, one or more rings can be cycloalkyl, aryl or heteroaryl, only if the connecting sites pass through the nonaromatic rings. In one example, nitrogen and/or sulfur atoms of a heterocyclyl radical group are selectively oxidized to provide N-oxide, sulfinyl and sulfonyl.

As used herein, the term "substituted heterocyclic"/"substituted heterocyclic alkyl"/"substituted heterocyclyl" refers to a heterocyclic group substituted by <NUM>-<NUM> (e.g. <NUM>-<NUM>) substituents which are the same as the substituent defined by the substituted cycloalkyl.

As used herein, the term "stereoisomer" refers to compounds with different chirality in stereocenters. Stereoisomers include enantiomers and diastereomers.

As used herein, the term "tautomer" refers to alternative forms of compounds with different proton locations, such as tautomer of enol ketone and imine enamine, or tautomeric forms of heteroaryl which contains ring atoms connected to the -NH- and =N- of the ring, such as pyrazol, imidazole, benzimidazole, triazole and tetrazole.

"Prodrug" refers to any derivative of the example compound, which can directly or indirectly provide the example compound, its active metabolite or residue when being applied to a subject. Particularly preferred derivatives and prodrugs are those that improve the bioavailability of the example compound (e.g. the compound administered orally tends to be absorbed in the blood more easily) or the delivery of the matrix compound to a biological compartment (such as brain or lymphocytic system) as per the matrix type when being applied to a subject. Prodrugs include ester forms in the compounds of the invention.

As used herein, the term "compounds in the invention" refers to compounds according to claim <NUM>, their racemes, stereoisomers or tautomers, or pharmaceutically acceptable salts.

The invention relates to: racemic mixtures of such compounds, mixtures enriched in any enantiomer, and separated enantiomer. The scope of the invention shall be understood the way that the racemic mixture refers to <NUM>% of two R and S enantiomers: <NUM>% of the mixture. The separated enantiomers shall be understood as pure enantiomers (i.e. <NUM>%) or mixtures with highly enriched enantiomers (purity ≥<NUM>%, ≥<NUM>%, ≥<NUM>% ^<NUM>%, ≥<NUM>%, ≥<NUM>% and ≥<NUM>%).

Where the compounds contain stereoisomers as mentioned in the invention, the invention shall include all the stereoisomers of such compounds.

Where the compounds contain tautomers as mentioned in the invention, the invention shall include all the tautomers of such compounds.

Also disclosed but not covered by the claimed invention are deuterated compounds generated from the replacement of any one/more hydrogen atoms in the compound with its/their stable isotope deuterium.

The invention also provides a pharmaceutical composition, which contains active ingredients at a safe and effective dosage, and pharmaceutically acceptable carriers.

The "active ingredient" in the invention refers to the compounds of the invention or their stereoisomers or tautomers, or pharmaceutically acceptable salts.

The "active ingredient" and the pharmaceutical composition in the invention can be used as IDO inhibitors. In another preferred embodiment, it is used to prepare drugs for the prevention and/or treatment of tumors. In another preferred embodiment, it is used to prepare drugs for the prevention and/or treatment of IDO mediated diseases.

"Safe and effective dosage" means: The dosage of active ingredient is sufficient to improve the condition without serious side effects significantly. Generally, the pharmaceutical composition contains <NUM>-<NUM>,<NUM> of the active ingredient/agent; preferably, it contains <NUM>-<NUM> of the active ingredient/agent. More preferably, "one dosage" is contained in a tablet.

"Pharmaceutically acceptable carrier" means: One or more compatible solid/liquid fillers or gel substances, which are suitable for human use, and shall be of sufficient purity and low toxicity. "Compatibility" here refers to the fact that each component of the composition can be blended with and among the active ingredients in the invention without significant reduction of the active ingredient's efficacy.

The compound of the invention can be administered as a single active agent or in combination with one or more other agents for cancer treatment. The combination of the compound of the invention with the known therapeutic agents and anticancer agents is also effective; the combination of the known compounds with other anticancer agents or chemotherapy agents is within the scope of the preferred embodiments. Examples of such agents can be seen in <NPL>. Based on the special properties of drugs and cancers involved, an ordinary technician in the field can identify the effective drug combinations. Such anticancer agents include (but are not limited to) the ones as follows: Estrogen receptor modulators, androgen receptor modulators, retinol receptor modulators, cytotoxic/cell growth inhibitors, anti-proliferation agents, isopentenyl protein transferase inhibitors, histone deacetylase (HDAC) inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, inhibitors of cell proliferation and survival signal, apoptosis inducers, interference cell cycle checkpoint , CTLA4 antibody, PD-<NUM> antibody, PD-L1 antibody, etc. The compounds of the invention are also effective when administered in combination with radiotherapy.

In general, the compounds of the invention will be administered in a therapeutically effective dosage and any acceptable mode via any medicament of a similar effect. The actual dosages of the compounds (i.e. active ingredients) in the preferred embodiments are determined based on numerous factors, such as the severity of diseases to be treated, age and relative health of a patient, efficacy of the compounds used, and route & form of application. The drug may be administered for times a day (once or twice preferably a day). All of such factors are taken into account by the attending physician.

For the purpose of the preferred embodiment, the therapeutically effective dosage can be a daily total dosage generally, for example, from <NUM>-<NUM>,<NUM>/kg weight for one time or times (preferably <NUM>-<NUM>/kg weight per day for a patient). Dosage unit composition can include its dosage factors to form a daily dosage. The dosage forms are chosen depending on various factors, such as administration mode and bioavailability of drug substances. In general, the compounds of the invention can be administered as a pharmaceutical composition through any of the routes as follows: Oral, systemic (e.g. transdermal, intranasal or suppository), or parenteral (e.g. intramuscular, intravenous or subcutaneous). The preferred method of administration is oral, whose convenient daily dosage can be adjusted as per the bitterness. The composition may be made in the forms of tablet, pill, capsule, semi-solid, powder, sustained-release preparation, solution, suspension, elixir, aerosol or any other appropriate composition. Another preferred administration mode of compounds of the invention is inhalation, which is an effective mode to deliver therapeutic agents directly to the respiratory tract (refer to <CIT> for example).

Pharmaceutically acceptable carriers or excipients include: Treatment agents, drug delivery modifiers and accelerators, such as calcium phosphate, magnesium stearate, talc, monosaccharide, disaccharide, starch, gelatin, cellulose, sodium methylcellulose, carboxymethyl cellulose, glucose, hydroxypropyl-B-cyclodextrin, polyvinylpyrrolidone, low-melting-point wax, ion exchange resin, and any combination of two or more of them. Liquid and semi-solid excipients can be selected from glycerol, propylene glycol, water, ethanol and various oils (including petroleum, animal oil, vegetable oil or synthetic oils, such as peanut oil, soybean oil, mineral oil and sesame oil). The preferred liquid carriers (in particular those for injectable solutions) include water, brine, glucose aqueous solution and ethylene glycol. Other pharmaceutically acceptable excipients are described in <NPL>).

As used herein, the term "pharmaceutically acceptable salt" refers to a non-toxic acid or alkaline-earth metal salt of a compound of the invention. Such salts can be prepared in situ while the final separation and purification of compounds of the invention, or via the reaction among proper organic/inorganic acids, alkalis, alkali/acid or functional groups. Representative salts include, but are not limited to: Acetate, adipate, alginate, citrate, aspartate, benzoate, benzene sulfonate, disulfate, butyrate, camphorate, camphorsulfonate, diglucosate, cypionate, lauryl sulfate, esilate, glucose heptanate, glycerophosphate, hemisulphate, enanthate, hexanoate, fumarate, hydrochloride, hydrobromate, hydroiodate, <NUM>-hydroxyethyl sulfonate, lactate, maleate, mesylate, nicotinate, <NUM>-naphthyl sulfonate, oxalate, dihydroxynaphthalate, pectinate, thiocyanate, <NUM>-phenylpropionate, picrate, pivalate, propionate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate and undecanoate. In addition, the basic groups containing nitrogen can be quaternary-ammonium salted with the agents as follows: Alkyl halides such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl groups; dialkyl sulfates such as dimethyl, diethyl, dibutyl and diamyl sulfate; long-chain halides such as chlorides, bromides and iodides of decyl, lauryl, myristyl and alkyl; aromatic alkyl halides such as benzyl and phenylethyl bromide. Water soluble, oil soluble or dispersible products are obtained. Examples of acids that can be used to form pharmaceutically acceptable acid-addition salts include inorganic acids of hydrochloride, sulfuric acid, phosphoric acid, etc. as well as the organic acids of oxalic acid, maleic acid, methanesulfonic acid, succinic acid, citric acid, etc. Alkali-addition salts can be prepared in situ while final separation and purification of compounds of the invention, or via the reaction of carboxylic acid portion with proper alkali (such as pharmaceutically acceptable hydroxides of metal cations, carbonate or bicarbonate), ammonia, organic primary, secondary or tertiary amines, respectively. Pharmaceutically acceptable salts include, but are not limited to, alkali metal and alkaline-earth metal based cations, such as sodium, lithium, potassium, calcium, magnesium, aluminum salts, as well as non-toxic ammonium, quaternary, and amine cations, including, but not limited to: Ammonium, tetramethyl-ammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc. Other representative organic amines used to produce alkali-addition salts include diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc..

As used herein, the term "pharmaceutically acceptable prodrug" refers to the prodrug of the compound in the preferred embodiment, which converts rapidly in vivo to the matrix compound as shown in the above Formula, such as hydrolysis in blood. <NPL>) by <NPL> provide a complete discussion.

The invention will be further construed in combination with specific examples. It is noteworthy that such examples are intended only to construe the invention rather than to limit its scope. The experimental methods without specific conditions in the following examples are usually adopted as per conventional conditions (for example,<NPL>), or those recommended by the manufacturers. Percentages and numbers shall be counted by weight unless otherwise stated.

All the professional and scientific terms used in this paper share the same meaning as those familiar to the skilled in the field unless otherwise defined. In addition, any method and material similar to the content described in the paper can be applied to the methods in the disclosure. The preferable implementation methods and materials described in this paper are only for demonstration.

Lithium aluminum hydride (<NUM>, <NUM> mmol) was added into a three-necked flask containing tetrahydrofuran (<NUM>,<NUM>) in batches. Tetrahydrofuran solution (<NUM>) of diisopropyl <NUM>,<NUM>-dimethoxycyclobutane-<NUM>,<NUM>-dicarboxylate (<NUM>, <NUM> mmol) was added slowly. After stirring at room temperature for <NUM>, TLC showed that the reaction had was completed. Saturated potassium sodium tartrate solution (<NUM>) was added to quench the reaction in ice bath, and the mixture was stirred at room temperature for <NUM>. The mixture was filtered and the filter cake was washed with dichloromethane/methanol (<NUM>:<NUM>, <NUM> x <NUM>), and the combined organic phase was concentrated to obtain colorless oily product (<NUM>, yield: <NUM>%).

<NUM>HNMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

The product of Step <NUM> (<NUM>, <NUM> mmol) was dissolved in pyridine (<NUM>), cooled in ice bath and treated with p-toluenesulfonyl chloride (<NUM>, <NUM> mmol) in batches. After stirring for <NUM> at room temperature, TLC showed that the reaction was completed. The reaction solution was filtered and then poured slowly into water (<NUM>). The solid was collected by filtration to obtain the product as a white solid (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (d, <NUM>).

N,N-dimethylformamide (<NUM>) was added into a three-necked flask, followed by sodium hydride (<NUM>, <NUM>. 8mmol) in batches. Nitrogen was changed for three times. Diisopropyl malonate (<NUM>, <NUM>. 1mmol) was added dropwise into the solution and stirred at room temperature for <NUM>. A solution of the product of Step <NUM> (<NUM>, <NUM> mmol) and KI (<NUM>, <NUM> mmol) in N, N-dimethylformamide (<NUM>) was added into the reaction system, and stirred at <NUM> for <NUM>. TLC showed that the reaction was completed. The cooled reaction solution was poured into water (<NUM>,<NUM>), extracted and concentrated with petroleum ether (<NUM> x <NUM>) and distilled under reduced pressure (<NUM>-<NUM>/<NUM> mmHg) to obtain a light-yellow oily product (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (sep, <NUM>).

The product of Step <NUM> (<NUM>, <NUM> mmol) and hydrochloric acid (<NUM>, <NUM>) were placed in a round-bottom flask and stirred at room temperature for <NUM>. After the reaction had been completed, white solid were generated which was collected by filtration to give the title product (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (sep, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> and dried tetrahydrofuran were added into a three-necked flask. Nitrogen was filled for three times. The solution was cooled to -<NUM> in dry-ice acetone bath. NaHMDS tetrahydrofuran solution (<NUM>, <NUM>, <NUM> mmol) was slowly added into the solution and stirred under the protection of nitrogen at -<NUM>. N-phenylbis(trifluoromethanesulfonimide) solution (<NUM>, in <NUM> of tetrahydrofuran) was slowly added into the solution and stirred at -<NUM> for <NUM> under N<NUM>. Saturated ammonium chloride solution (<NUM>) was added, and the mixture was extracted with <NUM> x <NUM> EtOAc. The combined organic phases were washed with saturated salt solution (<NUM>) and dried over sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>: <NUM>-<NUM>: <NUM>) to obtain the target product as a colorless oil (<NUM>, <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM> (sep, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (d, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>-quinolinyl-boronic acid (<NUM>, <NUM> mmol) and potassium carbonate (<NUM>, <NUM> mmol) were placed in a flask, followed by <NUM> of dioxane and tetrakis(triphenylphosphine)palladium (<NUM>, <NUM> mmol). Nitrogen was filled for three times and the reaction mixture was stirred at <NUM> for <NUM> under N<NUM> till the reaction had been completed. The reaction mixture (untreated) was directly used in next step without further purification.

Palladium-carbon catalyst (<NUM>, <NUM>% Pd/C) and methanol (<NUM>) were added into the reaction mixture of Step <NUM> and stirred at room temperature for <NUM> in hydrogen atmosphere till the reaction had been completed. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with dichloromethane: methanol (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the target product as a yellow oil (<NUM>, <NUM>% yield in two steps).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM>-<NUM> (m,<NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (sep, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, <NUM>).

The product of Step <NUM> (<NUM>, <NUM> mmol) was dissolved in ethanol (<NUM>) and treated with sodium hydroxide (<NUM>, <NUM>). The reaction mixture was heated to <NUM> and stirred for <NUM>. Hydrochloric acid (<NUM>) was added into the solution till the pH of the mixture was <NUM>. The mixture was concentrated to obtain the crude product (<NUM>).

A solution of the crude product (<NUM>) of Step <NUM> in pyridine (<NUM>) was heated to reflux <NUM>. Hydrochloric acid (<NUM>) was added into the solution until the pH of the mixture was <NUM>. The mixture was concentrated to obtain the crude product (<NUM>).

A mixture of the crude product (<NUM>) of Step <NUM>, triethylamine (<NUM>, <NUM> mmol), N,N-dimethylformamide (<NUM>) and <NUM>-(<NUM>-oxide benzotriazole)-N,N,N',N'-HATU (<NUM>, <NUM> mmol) was stirred at room temperature for <NUM>. p-Chloroaniline (<NUM>, <NUM> mmol) was the added into the mixture and stirred at room temperature for <NUM>. Ethyl acetate (<NUM>) was added, and the mixture was washed with water (<NUM>) for three times. The aqueous phase was extracted with ethyl acetate (<NUM> × <NUM>. The combined organic phase was washed with brine (<NUM>), dried with sodium sulfate, filtered and concentrated. The residual was purified by silica gel column chromatography eluted with dichloromethane: methanol (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the crude product (<NUM>). The product was further purified by pre-TLC to provide the title product as a yellow solid (<NUM>; <NUM>% in three steps ).

MS ESI: m/z =<NUM>, [M+H] +. <NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM>-<NUM> (m,<NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Sodium hydride (<NUM>, <NUM> mmol) and triethyl phosphonoacetate (<NUM>, <NUM> mmol) were dissolved in N, N-dimethylformamide (<NUM>) in ice bath. After stirring for <NUM>, a solution of <NUM>, <NUM>-dioxaspiro [<NUM>] decan-<NUM>-ketone (<NUM>, <NUM> mmol) in N, N-dimethylformamide solution (<NUM>), was added. After stirring at room temperature for <NUM>, the mixture quenched with water (<NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain the title product as an oil (<NUM>, <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM> (t, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

A solution of trimethylsulfonium iodide (<NUM>, <NUM> mmol) and potassium tert-butoxide (<NUM>, <NUM> mmol) in DMSO (<NUM>) was stirred at room temperature for half an hour. The product of Step <NUM> (<NUM>, <NUM> mmol) in DMSO (<NUM>) was added. After stirring at room temperature for <NUM> days, the mixture was quenched with water (<NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain the product as an oil (<NUM>, <NUM>%).

To a solution of the product (<NUM>, <NUM> mmol) of Step <NUM> in tetrahydrofuran (<NUM>) was added HCl (<NUM>, <NUM>). After stirring overnight, the mixture was quenched with buffer solution (pH=<NUM>, <NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>: <NUM>) to obtain the product as an oil (<NUM>, <NUM>%).

A solution of N-phenyl-bis(trifluoromethane)sulfonimide (<NUM>, <NUM> mmol) and NaHMDS (<NUM>, <NUM>, <NUM> mmol) in tetrahydrofuran (<NUM>) was stirred at -<NUM> under N<NUM> for <NUM>. The product of Step <NUM> (<NUM>, <NUM> mmol) was added into the solution. After stirring for <NUM>, the mixture was quenched with sodium posphate buffer solution (pH=<NUM>, <NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain the product as an oil (<NUM>, <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (t, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> and quinoline-<NUM>-boric acid (<NUM>, <NUM> mmol) were dissolved in tetrahydrofuran (<NUM>). Potassium carbonate (<NUM>, <NUM> mmol) and tetrakis(triphenylphosphine)palladium (<NUM>, <NUM>%) were added into the solution. After at <NUM> for <NUM> under N<NUM>, the solid was filtered off and the filtrate was treated with buffer salt (pH=<NUM>, <NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine and dried with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain the product as a yellow oil (<NUM>, <NUM>%).

To a solution of the product of Step <NUM> (<NUM>, <NUM> mmol) in ethanol (<NUM>) was added palladium-carbon catalyst (<NUM>, <NUM>% Pd/C). The reaction mixture was stirred under H<NUM> overnight. After filtering the palladium-carbon catalyst and concentrating the solution, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain the product as a white solid (<NUM>, <NUM>%).

The product (<NUM>, <NUM> mmol) of Step <NUM> was dissolved in tetrahydrofuran/ethanol (<NUM>/<NUM>)and treated with aqueous lithium hydroxide solution (<NUM>, <NUM>). After stirring at <NUM> for <NUM>, the mixture was quenched with phosphate buffer solution (pH=<NUM>, <NUM>) and extracted with ethyl acetate (<NUM> x <NUM>). The combined organic layer was washed with brine, filtered over anhydrous sodium sulfate and concentrated. The residual was purified by gel column chromatography to obtain two isomers: Isomer A (<NUM>) and isomer B (<NUM>).

The Isomers A (<NUM>, <NUM> mmol) obtained in Step <NUM>, <NUM>-chloroaniline (<NUM>, <NUM> mmol), <NUM>-(<NUM>-oxide benzotriazole)-N, N, N', N'-HATU (<NUM>, <NUM> mmol) and diisopropylethylamine (<NUM>µL, <NUM> mmol) were dissolved in dichloromethane (<NUM>). The resulted solution was stirred at room temperature for <NUM>, and then quenched with phosphate buffer solution (pH=<NUM>, <NUM>) and extracted with ethyl acetate (<NUM> mLx3). The combined organic layer was washed with brine and dried with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>) to obtain Example 2A as a white solid (<NUM>, <NUM>%).

Similarly, Example 2B was obtained from Isomer B of Step <NUM> as a white solid (<NUM>, <NUM>%). MS ESI: m/z = <NUM>, [M+H] +.

Tert-butyllithium (<NUM> <NUM> mol/L pentane solution, <NUM> mmol) was slowly added into a of <NUM>-bromine-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol) in <NUM> of THF at -<NUM> under argon. After stirring for <NUM>, a solution of the product (<NUM>, <NUM> mmol) of step <NUM> in Example <NUM> in THF (<NUM>) was added dropwise, and the mixture was stirred for <NUM> and slowly warmed to the room temperature. After acidifying the mixture with acetic acid (<NUM>) and concentrating, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product as a yellow oil (<NUM>, yield: <NUM>%).

The product (<NUM>, <NUM> mmol) of Step <NUM> and red phosphorus (<NUM>, <NUM> mmol) were mixed in hydroiodic acid (<NUM>, <NUM>%). The mixture was stirred at <NUM> for <NUM> in a sealed tube, then heated at <NUM> for <NUM>. After cooling, to room temperature, red phosphorus was filtered off, and sodium carbonate (<NUM>) was added to neutralize the mixture. Sodium thiosulfate pentahydrate (<NUM>) was added to remove the iodine. Sodium dihydrogen phosphate dihydrate (<NUM>) was added to adjust the pH value. The mixture was extracted with ethyl acetate (<NUM> x <NUM>) for three times. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate and concentrated. The residual was purified by silica gel column chromatography eluted with DCM:MeOH (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product as a white solid (<NUM>, yield: <NUM>%).

The product (<NUM>) of Step <NUM>, triethylamine (<NUM>, <NUM> mmol), N,N-dimethylformamide (<NUM>) and HATU (<NUM>, <NUM> mmol) were added into a flask. After stirring for <NUM>, p-chloroaniline (<NUM>, <NUM> mmol) was further added, and the mixture was stirred at room temperature for <NUM>. Ethyl acetate (<NUM>) was added, and the solution was washed with water (<NUM>) for three times. The aqueous phase was extracted with ethyl acetate (3x10mL). The combined organic phase was washed with brine (<NUM>), dried over sodium sulfate, filtered and concentrated. The residual was purified by silica gel column chromatography eluted with DCM:methanol (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the crude product (<NUM>) which was further purified by perp-TLC to give the product as a yellow solid (<NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (s, <NUM>), <NUM>-<NUM> (m,<NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

<NUM>,<NUM>-Cyclohexanedione monoethylene ketal (<NUM>, <NUM> mmol) and N-phenylbis (trifluoromethanesulfonimide) (<NUM>, <NUM> mmol) were dissolved in tetrahydrofuran (<NUM>). The mixture was cooled to -<NUM> and treated with a solution of NaHMDS solution (<NUM>, <NUM> in THF, <NUM> mmol) over <NUM>. After stirring for <NUM>, brine (<NUM>) was added, and the reaction solution was concentrated. Ethyl acetate (<NUM>) was added and the organic layer was washed with <NUM>% sodium hydroxide solution (<NUM>) twice. Ethyl acetate was concentrated to obtain the product (<NUM>; yield: <NUM>%), which was used for the next step without further purification.

<NUM>NMR (<NUM>, CDCl<NUM>): δ <NUM> (t, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (t, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>,<NUM>,<NUM>',<NUM>',<NUM>,<NUM>,<NUM>',<NUM>'-octamethyl-<NUM>,<NUM>'-bi(<NUM>,<NUM>,<NUM>-dioxaborolane) (<NUM>, <NUM> mmol), potassium acetate (<NUM>, <NUM> mmol), sodium bromide (<NUM>, <NUM> mmol) and [<NUM>,<NUM>'-bis (diphenylphosphino) ferrocene] dichloropalladium (<NUM>,<NUM> mmol) were added into a two-necked flask containing <NUM>,<NUM>-dioxane (<NUM>) under N<NUM>. The mixture was stirred at <NUM> for <NUM>. After concentration, ethyl acetate (<NUM>) was added, and the insoluble substance was filtered off. The residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product as a light yellow oil (<NUM>, <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM> (t, <NUM>), <NUM> (s, <NUM>).

The product (<NUM>,<NUM> mmol) of Step <NUM>, <NUM>-bromine-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol), potassium carbonate (<NUM>, <NUM> mmol) and tetrakis(triphenylphosphine) palladium were added into a two-necked flask containing <NUM>,<NUM>-dioxane (<NUM>) and water (<NUM>) under N<NUM>. The mixture was stirred at <NUM> for <NUM>. After the reaction mixture was concentrated, ethyl acetate (<NUM>) was added, and the insoluble substance was filtered off. The residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain <NUM> of the product (yield: <NUM>%).

<NUM>H NMR(<NUM>, CDCl<NUM>): δ <NUM>-<NUM> (d, <NUM>), <NUM>-<NUM> (q, <NUM>), <NUM>-<NUM> (q, <NUM>), <NUM>-<NUM> (td, <NUM>), <NUM>-<NUM>(d,<NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-l. <NUM>(t, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> was placed in a two-necked flask that contained isopropanol (<NUM>). Palladium-carbon catalyst (<NUM>, <NUM>%) was added, and the reaction mixture was heated to <NUM> °e under the atmosphere of hydrogen for <NUM>. The reaction mixture was filtered and concentrated to obtain <NUM> of the crude product which was used for the next step without further purification.

The product (<NUM>, <NUM> mmol) of Step <NUM> was placed in a single-neck bottle that contained acetone (<NUM>). Hydrochloric acid (<NUM>, <NUM>) was added and the mixture was attired at <NUM> for <NUM>. The reaction mixture was concentrated and ethyl acetate (<NUM>) and water (<NUM>) were added. The pH value of the mixture was adjusted to <NUM> with saturated sodium bicarbonate aqueous solution. The organic phase was separated and the aqueous phase was further extracted with ethyl acetate (<NUM> x <NUM>). The combined ethyl acetate phase was dried with anhydrous sodium sulfate. After filtration and concentration, the residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain <NUM> of the product (yield: <NUM>%).

To a suspension of NaH (<NUM>, <NUM> mmol) in <NUM> of N,N-dimethylformamide cooled in an ice bath under N<NUM> was added slowly a solution of triethylphosphonoacetate (<NUM>, <NUM> mmol in <NUM> of N,N-dimethylformamide), followed by addition of a solution of the product (<NUM>, <NUM> mmol) of Step <NUM> in <NUM> of N,N-dimethylformamide. After the reaction mixture wad stirred at room temperature for <NUM>, N,N-Dimethylformamide was evaporated and <NUM> of phosphate buffer solution (pH=<NUM>) was added. The mixture was extracted with ethyl acetate (<NUM> × <NUM>). The combined organic phase was washed with brine (<NUM>) and dried with sodium sulfate. After filtration and concentration, the residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the target product.

Potassium tert-butoxide (<NUM>, <NUM> mmol) and trimethylsulfonium iodide (<NUM>, <NUM> mmol) were dissolved in <NUM> of dimethyl sulfoxide at room temperature. After stirring overnight, a solution of the product of Step <NUM> (<NUM>, in <NUM> of DMSO) was added and the reaction mixture was stirred for <NUM> days at room temperature. The reaction was quenched with <NUM> of water and the mixture was extracted with ethyl acetate (<NUM> x <NUM>). The organic phases were washed with brine (<NUM>) and dried with sodium sulfate. After filtration and concentration, the residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product.

To a solution of the product of Step <NUM> (<NUM>, <NUM> mmol) in <NUM> of ethyl alcohol at room temperature was added <NUM> of LiOH solution (<NUM>, 1N). After stirring at <NUM> for <NUM>, water (<NUM>) was added and the mixture was extracted with ethyl acetate (<NUM> x <NUM>). The organic phase was washed with brine (<NUM>) and dried with sodium sulfate. After filtration and concentration, the residue was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product.

According to the method described in Example <NUM>, (±) (cis/trans) <NUM>-(quinolone-<NUM>-yl) spiro [<NUM>] octane-<NUM>-carboxylic acid in Step <NUM> was substituted by (±) (cis/trans) <NUM>-(<NUM>-fluoroquinoline-<NUM>-yl) spiro [<NUM>] octane-<NUM>-arboxylic acid to obtain the title product.

According to the method described in Example <NUM>, <NUM>-chloroaniline in Step <NUM> was substituted by <NUM>-fluoroaniline to obtain the title product.

According to the method described in Example <NUM>, the reagent <NUM>-chloroaniline in Step <NUM> was substituted by <NUM>-aminotrifluorotoluene to obtain the target compound.

According to the method described in Example <NUM>, the reagent <NUM>-chloroaniline in Step <NUM> was substituted by aniline to obtain the target compound.

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM>, diphenylphosphoryl azide (<NUM>µ L, <NUM> mmol) and triethylamine (<NUM>µ L, <NUM> mmol) were dissolved in toluene under N<NUM>. After stirring at <NUM> for <NUM>, the mixture was concentrated, and treated with water/tetrahydrofuran (<NUM>/<NUM>) and lithium hydroxide (<NUM>, <NUM> mmol). After stirring at room temperature for <NUM>, the reaction mixture was extracted with ethyl acetate (<NUM> x <NUM>). The combined organic phase was washed with brine (<NUM>), dried with sodium sulfate, filtered and concentrated to obtain the target product.

Starting with the product of Step1 and p-chlorobenzoic acid, the title product was obtained according to the method described in Example <NUM>.

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM> and triethylamine (<NUM>µL, <NUM> mmol) were dissolved in <NUM> of methylene chloride. A solution of <NUM>-chlorophenyl isocyanate (<NUM>, <NUM> mmol) in <NUM> of DCM was added. After stirring at room temperature for <NUM>, water (<NUM>) was added, and the mixture was extracted with ethyl acetate (<NUM> × <NUM>). The combined organic phase was washed with brine (<NUM>) and dried with sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether:ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the target product.

(<NUM>-Bromocyclopropyl)triphenylphosphonium bromide (<NUM>, <NUM> mmol) and sodium hydride (<NUM>, <NUM>. 68mmol) were placed in a two-necked flask under N<NUM>. Ultra-dry tetrahydrofuran (<NUM>) was added into a reaction flask, and the mixture was heated to <NUM> and stirred for <NUM>. A solution of the product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM> in tetrahydrofuran (<NUM>) was added into the reaction solution. After stirring for <NUM>, the reaction mixture was cooled to room temperature, and the insoluble substances were filtered off. After concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product (white solid, <NUM>; yield: <NUM>%).

<NUM>-Chloroperoxybenzoic acid (<NUM>, <NUM> mmol) was added to a solution of the product (<NUM>, <NUM> mmol) of Step <NUM> in dichloromethane (<NUM>) dissolved cooled in an ice bath. After stirring for <NUM>, methanesulfonic acid (<NUM>, <NUM> mmol) was added. After stirring at room temperature for <NUM>, the pH of the mixture was adjusted to <NUM> with saturated aqueous NaHCO<NUM> solution. The organic phase was washed with brine (<NUM> mLx2), dried with anhydrous sodium sulfate and concentrated. The residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the cis-trans isomers A-<NUM> (<NUM>) and B-<NUM> (<NUM>) (yield: <NUM>%). MS ESI: m/z =<NUM>, [M+H] +.

Potassium tert-butoxide (<NUM>, <NUM> mmol) was added into a solution of cis-trans isomers A-<NUM> (<NUM>, <NUM> mmol) produced in Step <NUM> in glycol dimethyl ether (<NUM>), followed by tosylmethyl isocyanide (<NUM>, <NUM> mmol) and methanol (<NUM>, <NUM> mmol) at <NUM>. After stirring at <NUM> oC for <NUM> and room temperature for <NUM>, saturated sodium bicarbonate solution (<NUM>) was added. The aqueous phase was extracted with glycol dimethyl ether. The combined organic phase was dried with anhydrous sodium sulfate and concentrated. The residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the cis-trans isomer A-<NUM> (<NUM>; yield: <NUM>%).

Starting with the cis-trans isomer A-<NUM>, cis-trans isomer B-<NUM> was prepared by the same method as for cis-trans isomer A-<NUM>.

The product A-<NUM> (<NUM>, <NUM> mmol) of Step <NUM> was added into hydrobromic acid (<NUM>%) (<NUM>), and the mixture was stirred at <NUM> for two days. The mixture was neutralized with saturated sodium bicarbonate solution till the pH was <NUM>, and extracted with ethyl acetate (<NUM> mLx3). The combined extracts were dried with anhydrous sodium sulfate, concentrated to obtain the product cis-trans isomer A-<NUM> (<NUM>; yield: <NUM>%).

Starting with the cis-trans isomer B-<NUM>, cis-trans isomer B-<NUM> was prepared by the same method as for cis-trans isomer A-<NUM>.

The cis-trans isomer A-<NUM> (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>-chloroaniline (<NUM>, <NUM> mmol), <NUM>-(<NUM>-azabenzotriazol-<NUM>-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (<NUM>, <NUM> mmol) and N,N-diisopropylethylamine (<NUM>, <NUM> mmol) were added into dichloromethane (<NUM>). The mixture was stirred at room temperature for <NUM>, and then at <NUM> for <NUM>. Water (<NUM>) was added, and the mixture was extracted with dichloromethane (<NUM> mLx3). The combined organic phase was dried with anhydrous sodium sulfate. The residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product (<NUM>; yield: <NUM>%).

Starting with the cis-trans isomer B-<NUM>, 10B was prepared by the same method as for cis-trans isomer 10A.

A3 (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM>, diphenylphosphoryl azide (<NUM>, <NUM> mmol) and toluene (<NUM>) of triethylamine (<NUM>, <NUM> mmol) were combined and heated to <NUM> under N<NUM>. After stirring for <NUM>, the mixture was concentrated and treated with water/tetrahydrofuran (<NUM>/<NUM>) and lithium hydroxide (<NUM>, <NUM> mmol) were added. After stirring for <NUM>, the mixture was concentrated. Ethyl acetate (<NUM>) was added, and the mixture was stirred for <NUM> and filtered. The filtrate was dried with anhydrous sodium sulfate and concentrated to obtain the crude product.

The product (<NUM>, <NUM> mmol) of Step <NUM> and chlorophenyl isocyanate (<NUM>, <NUM> mmol) were added into tetrahydrofuran (<NUM>), and the mixture was stirred at room temperature for <NUM>. Water (<NUM>) was added, and the mixture was extracted with ethyl acetate (<NUM> × <NUM>). The EtOAc extract was dried and concentrated, and the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product (<NUM>; yield: <NUM>%).

Starting with A-<NUM>, 11B was prepared by the same method described as for 11A.

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM> and triethylamine (<NUM>, <NUM> mmol) were dissolved in tetrahydrofuran (<NUM>) cooled in an ice bath. p-Chlorobenzoyl chloride (<NUM>, <NUM> mmol) was added into the reaction mixture. After stirring at room temperature for <NUM>, water (<NUM>) was added into reaction mixture, and the mixture was extracted with ethyl acetate (<NUM> x <NUM>). The combined organic phase was concentrated and the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product 12A as a white solid (<NUM>; yield: <NUM>%).

Starting with the product of Step <NUM> in Example 11B, 12B was prepared by the same method described as for 12A.

To a solution of the product (<NUM>, <NUM> mol) of Step <NUM> in Example 11A, <NUM>-dimethylaminopyridine (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol) in tetrahydrofuran (<NUM>) cooled in an ice bath was added <NUM>-chlorobenzenesulfonyl chloride (<NUM>, <NUM> mmol). After stirring at room temperature overnight, water (<NUM>) was added and the mixture was extracted with ethyl acetate (<NUM> × <NUM>). The combined organic phase was concentrated and the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the product as a white solid (<NUM>; yield: <NUM>%).

Starting with the product of Step <NUM> in Example 11B, 13B was prepared by the same method described as for 13A.

Starting with <NUM>-bromoaniline, 14A was prepared by the same method described as for 12A.

Starting with <NUM>-bromoaniline, 14B was prepared by the same method described as for 14A. MS ESI: m/z =<NUM>, [M+H] +.

To a suspension of methyltriphenylphosphonium bromide (<NUM>, <NUM> mmol) in diethyl ether (<NUM>) cooled at <NUM> as added potassium tert-butoxide (<NUM>, <NUM> mmol) in batches under N<NUM>. After stirring at room temperature for <NUM>, a ether solution (<NUM>) of <NUM>,<NUM>-cyclohexanedione monoethylene ketal was slowly added at <NUM>. The mixture was refluxed for <NUM> and then treated with water (<NUM>) and filtered. The filtrate was extracted with ethyl acetate (<NUM> × <NUM>). The combined organic phase was washed with brine (<NUM>), dried with anhydrous sodium sulfate and concentrated to give the title product as a brown oil (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (t, <NUM>), <NUM> (t, <NUM>), <NUM> (s, <NUM>), <NUM>(s, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> and Zn-Cu couple (<NUM>, <NUM> mmol) were suspended in methyl t-butyl ether (<NUM>) under N2. A glycol dimethyl ether solution (<NUM>) of trichloroacetic chloride was slowly added at <NUM>. After stirring at room temperature overnight, a methanol solution (<NUM>) of saturated ammonia chloride was added at <NUM> and the mixture was stirred at room temperature for <NUM>. The mixture was filtered through a pad of celite and filtrate was concentrated. The residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product as a white solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (t, <NUM>), <NUM> (t, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, tosylmethyl isocyanide (<NUM>, <NUM> mmol) and anhydrous ethanol (<NUM>, <NUM> mmol) were into a three-necked flask, followed by addition of t-butanol (<NUM>) and glycol dimethyl ether (<NUM>) cooled at <NUM>, followed by addition of potassium tert-butoxide (<NUM>, <NUM> mmol). After stirring at room temperature overnight, the reaction mixture was poured into ice water (<NUM>) and extracted with ethyl acetate (<NUM> × <NUM>). The EtOAc extract was washed with brine (<NUM>) and dried with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product as a white solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>(s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> was placed into a round-bottom flask, followed by addition of acetonitrile and water (<NUM>-<NUM>). The mixture was stirred at <NUM> for <NUM>. Water (<NUM>) was added and the acetonitrile was removed under reduced pressure. The mixture was extracted with ethyl acetate (<NUM> mLx2). The combined organic phase was washed with brine (<NUM>), dried with anhydrous sodium sulfate and concentrated. The residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> and tetrahydrofuran were added into a three-necked flask under N<NUM>. To the resulted solution cooled to -<NUM> was added slowly NaHMDS (<NUM>, <NUM> mmol, <NUM> in THF) under N<NUM>. After stirring for at -<NUM>, <NUM>, N-phenylbis (trifluoromethanesulfonimide) (<NUM> in <NUM> of THF) was added slowly and the mixture was stirred continuously for another hour at -<NUM>. Saturated ammonium chloride solution (<NUM>) was added and the mixture was extracted with 50mLx <NUM> ethyl acetate. The combined organic phase was washed with brine (<NUM>) and dried with sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product as an oil (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>-<NUM>(t, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, pinacol borate (<NUM>, <NUM> mmol) and potassium acetate (<NUM>, <NUM> mmol) were added into a flask, followed by addition of <NUM>,<NUM>-dioxane (<NUM>). and [<NUM>,<NUM>'-bis (diphenylphosphino)ferrocene]dichloropalladium (II) (<NUM>, <NUM> mmol) under N<NUM>. The reaction mixture was stirred at <NUM> for <NUM>, filtered through a pad of celite and concentrated. The residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(s, <NUM>), <NUM>(s, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>-bromo-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol), tetrakis(triphenylphosphine)palladium (<NUM>, <NUM> mmol) and cesium carbonate (<NUM>, <NUM> mmol) were added into dioxane (<NUM>) and water (<NUM>). The mixture was stirred at <NUM> overnight, filtered and concentrated. The residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain the product as a white solid.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>(d, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

A mixture of the product (<NUM>, <NUM> mmol) of Step <NUM>, water (<NUM>), isopropanol (<NUM>) and potassium hydroxide (<NUM>, <NUM> mmol) was stirred at <NUM> for <NUM>. Hydrochloric acid (<NUM> N) was added until the pH of the mixture was <NUM>. Ethanol (<NUM>) was added into the reaction mixture, followed by palladium-carbon catalyst (<NUM>) under N<NUM>. The mixture was stirred under H<NUM> for <NUM> and filtered through celite. The filtrate was concentrated o obtain the product as a white solid (<NUM>; yield: <NUM>%).

Racemic <NUM>: A solution of the product (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>-chloroaniline (<NUM>, <NUM> mol), N,N-diisopropylethylamine (<NUM>, <NUM> mmol) and HATU (<NUM>, <NUM> mmol) in DMF (<NUM>) was stirred at room temperature for <NUM>. The reaction mixture was poured into water (<NUM>) and extracted with ethyl acetate (<NUM>). The organic phase was washed with saturated sodium bicarbonate (<NUM> mLx2), brine (<NUM>), and dried with anhydrous sodium sulfate. After drying and concentration, the residual was purified by silica gel column chromatography eluted with a gradient of <NUM> to <NUM>% ethyl acetate/n-hexane to obtain racemic <NUM> (<NUM>; yield: <NUM>%).

Preparation of isomer 15A of Step <NUM>: <NUM> of racemic <NUM> was separated with chiral column AD-H (n-hexane: ethyl alcohol, <NUM>:<NUM>). Enantiomer 15A (<NUM>; yield: <NUM>%) has a retention time of <NUM>.

Preparation of isomer 15B of Step <NUM>: <NUM> of racemic <NUM> was separated with chiral column AD-H (n-hexane: ethyl alcohol, <NUM>:<NUM>). Enantiomer 15B (<NUM>; yield: <NUM>%) has a retention time of <NUM>.

Chiral resolution was performed with Agilent <NUM> semi-preparative liquid chromatograph (for axial chirality).

Chiral column: CHIRALPAK AD-H <NUM> x <NUM>; Flow rate: <NUM>/min; Detection wavelength: <NUM>; Collect two enantiomers.

Conditions: <NUM> sample of Example <NUM> was dissolved in <NUM> of methanol (injection volume: <NUM>; eluent: n-hexane: ethanol=<NUM>: <NUM> (volume ratio)); The retention times of two enantiomers are <NUM> (15A) and <NUM> (15B), respectively.

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM>, diphenylphosphoryl azide (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol) were dissolved into toluene (<NUM>) under N<NUM> and stirred at <NUM> for <NUM>. p-Chloroaniline (<NUM>, <NUM> mmol) was added and stirred for <NUM>. The residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the target product (<NUM>; yield: <NUM>%).

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM>, diphenylphosphoryl azide (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol) were dissolved into toluene (<NUM>) under the N<NUM> protection and stirred at <NUM> for <NUM>. After concentration, a mixture of <NUM> N HCl/THF (<NUM>/<NUM>) was added and the mixture was stirred at room temperature for <NUM>. Saturated sodium bicarbonate water solution (<NUM>) was added, and the mixture was stirred for <NUM> and extracted with ethyl acetate (<NUM> x <NUM>). Evaporation of EtOAc provided the crude product which was directly used for the next step without further purification.

The product (<NUM>, <NUM> mmol) of Step <NUM> and triethylamine (<NUM>, <NUM> mmol) were dissolved into tetrahydrofuran (<NUM>). p-Chlorobenzoyl chloride (<NUM>, <NUM> mmol) was added into the reaction mixture cooled in ice bath. After stirring for <NUM>, water (<NUM>) was added, and the mixture was extracted with ethyl acetate (<NUM> × <NUM>). The ethyl acetate phase was dried, concentrated and the residue was purified with prep-TLC separated (petroleum ether: ethyl acetate = <NUM>:<NUM>) to obtain the product (<NUM>).

Starting with p-bromoaniline, the title compound was prepared by the same method as described in Step <NUM> of Example <NUM>.

Starting with p-fluoroaniline, the title compound was prepared by the same method as described in Step <NUM> of Example <NUM>.

Starting with <NUM>-bromobenzoyl chloride, the title compound was prepared by the same method as described for Example <NUM>.

Starting with <NUM>-fluorobenzoyl chloride, the title compound was prepared by the same method as described for Example <NUM>.

t-Butyllithium (<NUM> <NUM> in pentane, <NUM> mmol) was slowly added into a THF solution of <NUM>-bromine-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol in <NUM> THF) at -<NUM> under argon. After stirring for <NUM>, a solution of the product Step <NUM> in Example <NUM> (<NUM>) in <NUM> of THF was slowly added. After stirring for <NUM>, saturated aqueous NH<NUM>Cl solution (<NUM>) was added and the mixture was extracted ethyl acetate (<NUM> × <NUM>). The organic phase was washed with brine (<NUM>) and dried with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate = (<NUM>:<NUM>-<NUM>:<NUM>) to obtain the target product as a brown solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>). <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> was added into <NUM> of <NUM> mol/L HCl. After stirring at room temperature for <NUM>, the mixture was extracted with ethyl acetate (<NUM> × <NUM>). The aqueous phase was treated with sodium carbonate aqueous solution to adjust the pH value to <NUM>, and was extracted with ethyl acetate (<NUM> x <NUM>). The organic phase was washed with brine (<NUM>), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the target product as a yellow solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>). A proton that can be exchanged by heavy water is not found by nuclear magnetism.

Starting with the product (<NUM>, <NUM> mmol) of Step <NUM>, the title product (<NUM>; yield: <NUM>%) was prepared as a yellow solid by using the same method as described in Step <NUM> of Example <NUM> A and 10B.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> and red phosphorus (<NUM>, <NUM> mmol) were mixed into <NUM> of concentrated hydroiodic acid (<NUM>%). After stirring at <NUM> for <NUM> under N<NUM>, red phosphorus was filtered off and the filtrate was treated with <NUM> of sodium carbonate and then with <NUM> of sodium thiosulfate pentahydrate to remove iodine. The pH value of the mixture was adjusted to <NUM> with addition of <NUM> N NaOH. The solid was collected by filtration washed with petroleum ether to obtain the title product as a yellow solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, DMSO-d<NUM>): δ <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Racemic <NUM>: The product (<NUM>, <NUM> mmol) of Step <NUM>, triethylamine (<NUM>, <NUM> mmol) and dichloromethane (<NUM>) were placed in a flask, followed by HATU (<NUM>, <NUM> mmol). After stirring for <NUM>, p-chloroaniline (<NUM>, <NUM> mmol) was added and the mixture was stirred at room temperature for <NUM>. After concentration, the residual was purified by silica gel column chromatography eluted with petroleum ether: ethyl acetate = (<NUM>:<NUM>-<NUM>:<NUM>) to obtain <NUM> of the crude product, which was washed <NUM> times with n-hexane to obtain the title compound as a yellow solid (<NUM>, yield: <NUM>%).

<NUM>H NMR (<NUM>, DMSO-d<NUM>): δ <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>. <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Preparation of Enantiomer 22A: <NUM> of Raceme <NUM> was separated with chiral column AD-H (n-hexane: isopropanol, <NUM>:<NUM>). The Enantiomer 22A (<NUM>; yield: <NUM>%) has a retention time of <NUM>.

<NUM>H NMR (<NUM>, CD<NUM>OD): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Preparation of Enantiomer 22B: <NUM> of Raceme <NUM> was separated with chiral column AD-H (n-hexane: isopropanol, <NUM>:<NUM>). The Enantiomer 22A (<NUM>; yield: <NUM>%) has a retention time of <NUM>.

Chiral resolution was performed with Agilent <NUM> semi-preparative liquid chromatograph.

Chiral column: CHIRALPAK AD-H <NUM>*<NUM>; Flow rate: <NUM>/min; Detection wavelength: <NUM>; two peaks were collected.

Conditions: <NUM> sample in Example <NUM> was dissolved into <NUM> of methanol (injection volume: <NUM>; eluent: n-hexane: isopropanol = <NUM>:<NUM> (volume ratio); The retention times of two enantiomers were <NUM> and <NUM>, respectively.

Starting with p-bromoaniline, the title compound was prepared by the same method as described for Example <NUM>.

<NUM>H NMR (<NUM>, CD<NUM>OD): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Starting with p-fluoroaniline, the title compound was prepared by the same method as described for Example <NUM>.

<NUM> NMR (<NUM>, CD<NUM>OD): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, <NUM>), <NUM> (dt, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM> was dissolved into THF (<NUM>), followed addition of by dicyclohexylcarbodiimide (<NUM>, <NUM> mmol) and <NUM>-dimethylaminopyridine (<NUM>, <NUM> mmol). After stirring at room temperature for <NUM>, (S)-<NUM>-phenylethanol (<NUM>, <NUM> mmol) was added into the reaction mixture. The mixture was stirred for <NUM> and the byproduct dicyclohexylurea was filtered off. The filtrate was concentrated and the residual was purified by silica gel column chromatography for purification elute with a gradient of <NUM>-<NUM>% ethyl acetate/n-hexane to obtain the title compound as a colorless oil (<NUM>; yield: <NUM>%).

Separation performed with Agilent <NUM> semi-preparative liquid chromatograph system.

Chiral column: DAICEL IC <NUM> x <NUM>, <NUM>; Flow rate: <NUM>/min; Eluent: n-hexane: Ethanol=<NUM>: <NUM> (volume ratio); Detection wavelength: <NUM>; two peaks were collected at <NUM> (isomer A) and <NUM> (isomer B) respectively.

The product of Step <NUM> (<NUM>) was separated under the above-mentioned conditions to provide isomer A (<NUM>) and isomer B (<NUM>).

Isomer A in Step <NUM> was dissolved in methanol (<NUM>). Under N<NUM>, palladium hydroxide carbon catalyst (<NUM>, <NUM>%) and pure water (<NUM>) were added and the mixture was stirred at room temperature under H<NUM> for <NUM>. Palladium catalyst was filtered off through celite. The filtrate was concentrated and the residual was purified by silica gel column chromatography for purification eluted with a gradient of <NUM>-<NUM>% methanol/ dichloromethane to obtain the title product as a white solid (<NUM>; yield: <NUM>%), [α]D<NUM>= -<NUM> (c = <NUM>, CHCl<NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM>, <NUM>-fluoroaniline (<NUM>, <NUM> mol), <NUM>-ethyl-(<NUM>-dimethyllaminopropyl) carbonyl diamide hydrochloride (<NUM>, <NUM> mmol) and <NUM>-hydroxybenzotriazole (<NUM>, <NUM> mmol) were added into dichloromethane (<NUM>). Triethylamine (<NUM>, <NUM> mmol) was then added, and the mixture was stirred at room temperature for <NUM> in an ice bath. The reaction mixture was poured into water (<NUM>) and extracted with ethyl acetate (<NUM>). The organic phase was washed with saturated sodium bicarbonate (<NUM> x <NUM>), brine (<NUM>) and dried with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with a gradient of <NUM>-<NUM>% ethyl acetate/n-hexane to obtain the target product (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>):<NUM><NUM>(d, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

Using p-bromoaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>):<NUM><NUM>(d, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(s,<NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

The produt (<NUM>, <NUM> mmol) of Step <NUM> in Example <NUM>, <NUM>-aminopyridine (<NUM>, <NUM> mol) and N-methylimidazole (<NUM>, <NUM> mmol) were mixed in MeCN (<NUM>), followed by addition of N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate (<NUM>, <NUM> mmol). The mixture was stirred at room temperature for <NUM> and was poured into water (<NUM>) and extracted with ethyl acetate (<NUM>). The organic phase was washed with saturated sodium bicarbonate (<NUM> x <NUM>) and brine (<NUM>) and dried it with anhydrous sodium sulfate. After filtration and concentration, the residual was purified by silica gel column chromatography eluted with a gradient of <NUM>-<NUM>% ethyl acetate/n-hexane to obtain the target product (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>(d, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>(s, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(td, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>),<NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

Using <NUM>-fluoro-<NUM>-(<NUM>-methyl-<NUM>H-pyrazol-yl) aniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>):<NUM><NUM>(d, <NUM>), <NUM>-<NUM>(dd, <NUM>),<NUM>(s, <NUM>), <NUM>(d, <NUM>), <NUM>-<NUM>(td, <NUM>), <NUM>-<NUM>(td, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(d, <NUM>), <NUM>(s, <NUM>), <NUM>(s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

Using <NUM>-chloroaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (s, <NUM>), <NUM> (ddd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (td, <NUM>), <NUM> (p, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-chloro-<NUM>-fluoroaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (ddd, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-fluoroaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-bromaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-aminopyridine and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-fluoro-<NUM>-chloroaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

Using <NUM>-trifluoromethyl phenylamine and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM> NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (ddd, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-fluoro-<NUM>-bromaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (t, <NUM>), <NUM> (s, <NUM>), <NUM> (p, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>),<NUM>-<NUM> (m, <NUM>).

Using <NUM>-(trifluoromethyl)aniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>),<NUM> (s, <NUM>), <NUM> (p, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>- <NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>, <NUM>-difluroaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM> (ddd, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-amino-<NUM>-chloropyridine and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (s, <NUM>), <NUM> -<NUM> (m, <NUM>), <NUM> (ddd, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (dt, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using [<NUM>,<NUM>'-biphenyl]-<NUM>-amine and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (t, <NUM>), <NUM> (t, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (m, <NUM>), <NUM>(m, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-chlorine-<NUM>-trifluoromethyl-aniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM> (dt, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>).

<NUM>H NMR (<NUM>, CDCl<NUM>) :δ <NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM> (t, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>- <NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>).

Using <NUM>-bromaniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>):δ <NUM> (d, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using <NUM>-aminopyridine and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>) δ <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>).

Using <NUM>-aminobenzonitrile and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM> (d, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (dd, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>).

Using aniline and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>):<NUM><NUM>(d, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>(d, <NUM>), <NUM>-<NUM>(td, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(t, <NUM>), <NUM>(s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

To a solution of <NUM>-bromine-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol) in THF (<NUM>) cooled to -<NUM> was added t-butyllithium solution (<NUM>, <NUM> in pentane, <NUM> mmol) slowly under argon. After stirring for <NUM>, a solution of <NUM>-oxo-<NUM>-azaspiro [<NUM>] nonane-<NUM>-carboxylate (<NUM>, <NUM> mmol) in <NUM> of THF was added. After stirring at -<NUM> for <NUM>, acetic acid (<NUM>) was added and the mixture was concentrated. The residual was purified by silica gel column chromatography eluted with a gradient of <NUM>-<NUM>% ethyl acetate/n-hexane to obtain the title product as a light yellow solid (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>):δ <NUM> (d, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(dd, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(d, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(s, <NUM>).

The product (<NUM>, <NUM> mmol) of Step <NUM> was placed into a flask, followed by addition of concentrate hydroiodic acid (<NUM>, <NUM>%) and red phosphorus (<NUM>, <NUM> mmol). After the mixture was stirred at <NUM>) for <NUM>, <NUM> N sodium hydroxide aqueous solution was added to adjust the pH value to <NUM> and the mixture was treated with Sodium sulfate aqueous solution (<NUM>). After stirring for <NUM>, the solution was extracted with dichloromethane (<NUM> x <NUM>). The combined organic phase was dried with anhydrous sodium sulfate, filtered and concentrated to obtain the crude product (<NUM>; yield: <NUM>%).

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>(d, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>(s, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>), <NUM>-<NUM>(m, <NUM>).

Using <NUM>-fluorophenyl isocyanate and the product of Step <NUM>, the title product was prepared by the same method as described for Example <NUM>.

Using <NUM>-chlorophenyl isocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

Using <NUM>-bromophenyl isocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

Using <NUM>-isocyanatobenzonitrile isocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described for Example <NUM>.

To a solution of <NUM>-bromine-<NUM>-fluoroquinoline (<NUM>, <NUM> mmol) and triisopropyl borate (<NUM>, <NUM> mmol) in <NUM> of THF cooled to -<NUM> was added t-butyllithium (<NUM>, <NUM> in heptane, <NUM> mmol) under N<NUM>. After stirring for <NUM>, saturated ammonium chloride solution (<NUM>) was added, and the mixture was extracted with <NUM> x <NUM> of ethyl acetate. The combined organic phase was washed with brine (<NUM>), dried with sodium sulfate, filtered and concentrated to obtain <NUM> of crude product which was washed with petroleum ether : ethyl acetate (<NUM>: <NUM>) to obtain the title compound as a yellow solid (<NUM>; yield: <NUM>%).

Using the product of Step <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

<NUM>H NMR (<NUM>, CDCl<NUM>): δ <NUM>-<NUM> (m, <NUM>), <NUM> (d, <NUM>), <NUM> (d, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM>-<NUM> (m, <NUM>), <NUM> (s, <NUM>).

To a solution of the product (<NUM>, <NUM> mmol) of Step <NUM> in <NUM> of dichloromethane was added <NUM> of <NUM>,<NUM>,<NUM>-trifluoroacetic acid, After stirring for <NUM>, the solution was concentrated to obtain the target product as a yellow solid.

The product (<NUM>, <NUM> mmol) of Step <NUM>, triethylamine (<NUM>, <NUM> mmol) and <NUM> of dichloromethane were placed in a flask. <NUM>-Chlorophenyl isocyanate (<NUM>, <NUM> mmol) was added and the mixture was stirred at room temperature for <NUM>, and concentrated. The residual was purified by silica gel column chromatography to obtain the target product. MS ESI: m/z =<NUM>, [M+H] +.

Using <NUM>-bromophenyl isocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

Using <NUM>-cyanophenylisocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

Using <NUM>-fluorophenyl isocyanate and the product of Step <NUM> of Example <NUM>, the title product was prepared by the same method as described in Step <NUM> for Example <NUM>.

Firstly, DO gene was amplified with PCR, and the amplified PCR product was recycled. Digestion (<NUM> under <NUM>), gel running and recycling were carried out on the pET28a plasmid (purchased from Shanghai Baomanbio Co. ) and IDO gel with two restriction enzymes (EcoR I and Xho I). T4 ligase was connected with the product overnight, added into DH5 α competence, placed on the ice for <NUM> and subjected to thermal shock at <NUM> for <NUM>. Bacteria coated plate was shaken to pick up monoclonal for PCR identification and sequencing; if all were accurate, it indicated that pET28a-IDO plasmid was established successfully.

BL21 which contained pET28a-IDO plasmid was shaken vigorously at <NUM> till OD<NUM> was <NUM>-<NUM>, added into hemin (final concentration: <NUM>) and <NUM> IPTG (isopropyl- β -D-IPTG) and induced under <NUM> for <NUM>; after induction, thallus were collected centrifugally at <NUM> and <NUM>,000rpm, washed with <NUM> PBS (pH <NUM>) and collected again centrifugally.

The collected thallus were suspended again with buffer solution (<NUM> PBS pH <NUM>). An appropriate amount of <NUM> x PMSF was added. The thallus were disrupted with cell disruption instrument (<NUM>,400bar) at <NUM> for three times. The split bacteria were centrifuged at <NUM>,<NUM> x g for <NUM>. The sediment was removed but supernatant was retained and filtered with a <NUM> film at <NUM>; the nickel column was balanced with lysis buffer (<NUM> PBS pH7. <NUM>) by <NUM> column volumes. The supernatant was loaded on the nickel column and washed with wash solution (<NUM> PBS pH7. <NUM>, <NUM> imidazole) by <NUM> column volume. Finally, protein was eluted with eluant (<NUM> PBS pH7. <NUM>, <NUM> imidazole); dialyzed for <NUM> with dialysis solution (<NUM> PBS pH7. <NUM>), concentrated, sub-packaged, quickly frozen with liquid nitrogen and stored at -<NUM> for backup.

Firstly, the compound was diluted by triple gradient. 1µL of each concentration was added into a <NUM>-well plate; 50µL of IDO Enzyme solution PBS (pH <NUM>; final concentration: <NUM>) was added: 25µL of substrate <NUM> (methylene blue, final concentration: <NUM>; catalase, final concentration: <NUM>. 2µg/L; PBS (pH <NUM>; final concentration: <NUM>) mixture and 25µL of substrate <NUM> (D-Trp, final concentration: <NUM>; sodium ascorbate, final concentration: <NUM>; PBS (pH <NUM>), final concentration <NUM>) mixture were added for starting the reaction. Finally, OD<NUM> nm reading was carried out for <NUM>.

Hela cells (80µE) was inoculated on a <NUM>-well plate (<NUM> x <NUM><NUM> for each well) to grow overnight. After the compound was diluted the next day, 1µL of diluent and 100µL of culture medium which contains human interferon γ (final concentration: 50ng/mL) were added into the <NUM>-well plate to make the final volume reach 200µL. After incubating for <NUM>, 80µL of supernate was transferred from each well to a new <NUM>-well plate. 10µL <NUM>. 1N trichloroacetic acid was added into each well for mixing and incubation at <NUM> for <NUM>, and IDO catalyzed N formyl-kynurenin into kynurenine. The reaction mixture was centrifuged at <NUM>,500rpm for <NUM>, 70µL of supernatant of each well was transferred to a new <NUM>-well plate and mixed with 100µL of <NUM>% (w/v) dimethylaminobenzaldehyde acetic acid solution. After placing for <NUM>-<NUM>, measurement was carried out at <NUM> with the SPECTRAmax i3 reader.

The test result of IDO enzyme and celllular inhibitory activity of compounds is shown as Table <NUM>.

The above result shows that the compound in the invention has inhibitory activity for IDO enzyme and cell.

Claim 1:
A compound selected from the group of
N-(<NUM>-fluorophenyl)-<NUM>-(<NUM>-fluoroquinolin-<NUM>-yl)spiro[<NUM>]nonane-<NUM>-carboxamide,
N-(<NUM>-chlorophenyl)-<NUM>-(<NUM>-fluoroquinoline-<NUM>-yl)-<NUM>-azaspiro [<NUM>] nonane-<NUM>-carboxamide,
N-(<NUM>-bromophenyl)-<NUM>-(<NUM>-fluoroquinoline-<NUM>-yl)-<NUM>-azaspiro [<NUM>] nonane-<NUM>-carboxamide,
N-(<NUM>-cyanophenyl)-<NUM>-(<NUM>-fluoroquinoline-<NUM>-yl)-<NUM>-azaspiro [<NUM>] nonane-<NUM>-carboxamide, and
N-(<NUM>-fluorophenyl)-<NUM>-(<NUM>-fluoroquinoline-<NUM>-yl)-<NUM>-azaspiro [<NUM>] nonane-<NUM>-carboxamide
or their stereoisomers or tautomers, or pharmaceutically-acceptable salts.