Combination therapy for treating cancer

Abstract

To provide a method for treating cancer patients. [Solution] The method comprises administering to a patient an effective amount of a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and an effective amount of an immunomodulator. The Specified also provides compositions and kits for carrying out the method described herein. In another embodiment, the method comprises administering to a patient an effective amount of a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and an effective amount of radiotherapy. JPEG2026048648000034.jpg44160

Description

[Technical Field]

[0001] This application claims the benefit of priority under § 119(e) of U.S. Provisional Patent Application No. 62 / 930,054, filed on 4 November 2019, which is incorporated herein by reference in its entirety for all purposes. [Background technology]

[0002] Arginase is a manganese metalloenzyme that catalyzes the conversion of L-arginine to urea and L-ornithine. Two isoforms exist: arginase 1 and arginase 2. Although L-arginine is not an essential amino acid, as it can be supplied through protein turnover in healthy adults, increased expression and secretion of arginase under various physiological and pathological conditions (e.g., pregnancy, autoimmune diseases, cancer) leads to decreased L-arginine levels. Immune cells, in particular, are sensitive to decreased L-arginine levels. Tumors evade the immune system using multiple immunosuppressive mechanisms. One such mechanism involves decreased L-arginine due to increased circulating arginase levels, increased arginase expression and secretion by tumor cells, and the recruitment of bone marrow-derived suppressor cells that express and secrete arginase. These factors work together to result in decreased L-arginine and an immunosuppressive phenotype in the tumor microenvironment. Pharmacological inhibition of arginase activity has been shown to reverse low L-arginine-induced immunosuppression in animal models. However, many proteins and pathways are involved in cancer, and research into these is progressing rapidly. Therefore, there is a need for novel cancer treatments for patients. [Overview of the project] [Means for solving the problem]

[0003] In some embodiments, the present disclosure relates to a method for treating a cancer patient, wherein the formula is (Ia) or (Ib): [ka] [wherein, n is 0 or 1; R is -H or -C(O)CH(R 1a )NHR 1b ; R 1a is selected from -H, -(C1-C6)alkyl, and CH2OR 1c ; R 1b is -H; or alternatively, R 1a and R 1b together with the atom to which they are attached form a 5-membered heterocyclic ring; R 1c is H or -CH3] administering to a patient an effective amount of a compound, or a pharmaceutically acceptable salt thereof, and an effective amount of an immunomodulatory agent.

[0004] In some embodiments, the present disclosure provides a method of treating a cancer patient comprising administering to the patient an effective amount of a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and an effective amount of radiation therapy. In some embodiments, the method further comprises administering to the patient an effective amount of an immunomodulatory agent.

[0005] In some embodiments, the radiation therapy is fractionated radiation therapy.

[0006] In some embodiments of formula (Ia) or (Ib), R 1 is -H or -C(O)CH(R 1a )NH2; R 1a is selected from -H or -(C1-C6)alkyl.

[0007] In some embodiments of formula (Ia) or (Ib), the compound is of formula (IIa) or (IIb):

Chemical formula

[0008] In some embodiments, the immunomodulator is an immune checkpoint inhibitor or an immunostimulator. In some embodiments, the immune checkpoint inhibitor is selected from CTLA-4 receptor inhibitors, PD-1 receptor inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, NKG2A receptor inhibitors, and combinations thereof. In some embodiments, the immunostimulator is a TLR3 agonist.

[0009] In some embodiments, the immune checkpoint inhibitor is an antibody or an antigen-binding fragment thereof.

[0010] In some embodiments, the immune checkpoint inhibitor is an anti-CTLA-4 receptor antibody, an anti-PD-1 receptor antibody, an anti-PD-L1 antibody, an anti-PD-L2 antibody, or an anti-NKG2A receptor antibody. In some embodiments, the immune checkpoint inhibitor is an anti-CTLA-4 receptor antibody, an anti-PD-L1 antibody, an anti-NKG2A receptor antibody, or a combination thereof.

[0011] In some embodiments, the immune checkpoint inhibitor is durvalumab, tremelimumab, monalizumab, or a combination thereof.

[0012] In some embodiments, cancer is breast cancer, bladder cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, gastrointestinal stromal cancer, gastroesophageal carcinoma, renal cell carcinoma, prostate cancer, liver cancer, colon cancer, pancreatic cancer, ovarian cancer, lymphoma (including non-Hodgkin lymphoma), cutaneous T-cell lymphoma, or melanoma. In some embodiments, cancer is a hematological malignancy including multiple myeloma, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), chronic myelomonocytic leukemia (CMML), and diffuse large B-cell lymphoma (DLBCL).

[0013] In some embodiments, a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, is administered sequentially, separately, or simultaneously with an immunomodulator.

[0014] In some embodiments, the Disclosure provides compounds of formula (Ia) or (Ib), or pharmaceutically acceptable salts thereof, for use in the treatment of cancer patients, which are administered to the patient sequentially, separately, or simultaneously with an immunomodulator.

[0015] In some embodiments, the present disclosure provides immunomodulators for use in the treatment of cancer, wherein an immune checkpoint inhibitor is administered to the patient sequentially, separately, or simultaneously with a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof. [Brief explanation of the drawing]

[0016] [Figure 1] Figures 1A, 1B, 1C, and 1D show the time-dependent reduction in tumor volume in the MC38-ova trial with arginase inhibitors (Figures 1A and 1B), anti-PDL1 (Figures 1A and 1C), and a combination of arginase inhibitors (ARG inh) and anti-PDL1 (Figures 1A and 1D). [Figure 2] Figures 2A, 2B, 2C, and 2D show the time-dependent reduction in tumor volume in the MC38-ova trial with an arginase inhibitor (Figure 2A), a combination of an arginase inhibitor and anti-PDL1 (Figure 2B), a combination of an arginase inhibitor and anti-NKG2A (Figure 2C), and a combination of an arginase inhibitor, anti-PDL1, and anti-NKG2A (Figure 2D). [Figure 3A] Figure 3A shows the time-dependent decrease in vehicle tumor volume in the MC38-ova trial. [Figure 3B] Figure 3B shows the time-dependent reduction in tumor volume with arginase inhibitors in the MC38-ova trial. [Figure 3C]Figure 3C shows the time-dependent reduction in tumor volume with a TLR3 agonist in the MC38-ova trial. [Figure 3D] Figure 3D shows the time-dependent reduction in tumor volume with the combination of an arginase inhibitor and a TLR3 agonist in the MC38-ova trial. [Figure 3E] Figure 3E shows the time-dependent reduction in tumor volume in the MC38-ova trial with arginase inhibitors, TLR3 agonists, and combinations of arginase inhibitors and TLR3 agonists. [Figure 3F] Figure 3F shows the time-dependent reduction in tumor volume in the MC38-ova trial with arginase inhibitors, TLR3 agonists, and combinations of arginase inhibitors and TLR3 agonists. [Figure 4A] Figure 4A shows the changes in immune cells in tumors induced by arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 4B] Figure 4B shows the changes in immune cells in tumors induced by arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 4C] Figure 4C shows the changes in immune cells in tumors induced by arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 4D] Figure 4D shows the increased CD8+ (Figures 4A and 4B) and CD103+ (Figure 4F) T cell function in tumor inflow lymph nodes with arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 4E] Figure 4E shows the increased CD8+ (Figures 4A and 4B) and CD103+ (Figure 4F) T cell function in tumor inflow lymph nodes with arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 4F] Figure 4F shows the increased CD8+ (Figures 4A and 4B) and CD103+ (Figure 4F) T cell function in tumor inflow lymph nodes with arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 5] Figures 5A and 5B show the increase in IFNγ (Figure 5A) and TNFα (Figure 5B), which produce CD8+ T cell function, in tumor inflow lymph nodes induced by arginase inhibitors, anti-PDL1, and combinations of arginase inhibitors and anti-PDL1. [Figure 6A] Figure 6A shows the antitumor activity of the combination of an arginase inhibitor (compound 12) and radiotherapy (RT) in a Lewis lung syngenic tumor model. [Figure 6B] Figure 6B shows that the combination of an arginase inhibitor (compound 12) and radiotherapy (RT) reduced the volume of Lewis lung tumors at the end of the study (day 19). [Modes for carrying out the invention]

[0017] This disclosure relates to a method for treating cancer patients. In one embodiment, the method comprises administering to a patient a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, in combination with an immunomodulator. Compounds of formula (Ia) or (Ib), or any subgenus or species thereof, are useful in therapy as arginase inhibitors.

[0018] Unless specifically modified by the term "intact," as in "intact antibody," the term "antibody" as used herein also includes antibody fragments such as Fab, F(ab')2, Fv, scFv, Fd, dAb, and other antibody fragments that retain antigen-binding function, such as the ability to bind to antigens such as CTLA-4, PD1, PD-L1, or NKG2A. Typically, such fragments will contain an antigen-binding domain.

[0019] The term "mAb" refers to a monoclonal antibody. The antibodies of this disclosure may include, but are not limited to, whole-natural antibodies, bispecific antibodies; chimeric antibodies; Fab, Fab', single-chain V region fragments (scFv), fusion polypeptides, and non-conventional antibodies.

[0020] The terms “to treat,” “to treat,” and “treatment” include reducing or inhibiting arginase or cancer-related enzyme activity or protein activity in the subject, or remission of one or more symptoms of cancer in the subject, or slowing or delaying the progression of cancer in the subject. The terms “to treat,” “to treat,” and “treatment” also include reducing or inhibiting tumor growth or proliferation of cancer cells in the subject.

[0021] The terms “inhibit,” “inhibit,” or “the act of inhibiting” include a reduction in the baseline activity of a biological activity or biological process.

[0022] The term “pharmaceutical composition” includes compositions comprising an active ingredient, a pharmaceutically acceptable excipient, carrier, or diluent, wherein the active ingredient is a compound of formula (Ia) or (Ib) (including any subgenus or species thereof), a pharmaceutically acceptable salt thereof, or an immunomodulator described herein. The term “pharmaceutically acceptable excipient, carrier, or diluent” includes compounds, substances, compositions, and / or formulations suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, within the bounds of reasonable medical judgment as can be ascertained by those skilled in the art. In some embodiments, the pharmaceutical composition is in the form of a solid dosage form such as capsules, tablets, granules, powders, or sachets. In some embodiments, the pharmaceutical composition is in the form of an injectable sterile solution in one or more aqueous or non-aqueous, non-toxic, parenterally acceptable buffer systems, diluents, solubilizers, co-solvents, or carriers. Sterile injection preparations may also be sterile aqueous or oily suspensions for injection, or suspensions in non-aqueous diluents, carriers, or co-solvents, which may be formulated using one or more suitable dispersants or wetting agents and suspending agents according to known procedures. Pharmaceutical compositions may be solutions for IV bolus / infusion or lyophilized systems (alone or with excipients) for reduction with or without other excipients. Lyophilized substances may be prepared from non-aqueous or aqueous solvents. The dosage form may also be a concentrate for further dilution for subsequent infusions.

[0023] The term “patient” includes warm-blooded mammals, including, for example, primates, dogs, cats, rabbits, rats, and mice. In some embodiments, the subject is a primate, such as a human. In some embodiments, the patient has cancer. In some embodiments, cancer is breast cancer, bladder cancer, head and neck cancer, non-small cell lung cancer (NSCLC), small cell lung cancer, colorectal cancer, gastrointestinal stromal cancer, gastroesophageal carcinoma, renal cell carcinoma, prostate cancer, liver cancer, colon cancer, pancreatic cancer, ovarian cancer, lymphoma (including non-Hodgkin lymphoma), cutaneous T-cell lymphoma, or melanoma. In some embodiments, cancer is a hematological malignancy, including multiple myeloma, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), chronic myelomonocytic leukemia (CMML), and diffuse large B-cell lymphoma (DLBCL).

[0024] The phrase “effective amount” includes a compound of formula (Ia) or (Ib) (including all subgenus or species thereof) and / or an amount thereof of the immunomodulator described herein that would induce a biological or medical response in a subject (e.g., a decrease or inhibition of arginase or cancer-related enzyme or protein activity, remission of cancer symptoms, or slowing or delaying cancer progression). In some embodiments, the phrase “effective amount” includes a compound of formula (Ia) or (Ib) (including all subgenus or species thereof) and / or an amount thereof of the immunomodulator described herein that would be effective in at least partially mitigating, inhibiting and / or remitting cancer, or inhibiting arginase, and / or reducing or inhibiting tumor growth or proliferation of cancer cells in a subject.

[0025] compound In one embodiment, formula (Ia) [ka] [wherein n is 0 or 1; R 1 is -H or -C(O)CH(R1a )NHR 1b and; R 1a These are -H, -(C1~C6) alkyl, and CH2OR 1c Selected from; R 1b is -H; or alternatively, R 1a and R 1b These, together with the atoms they bond to, form a five-membered heterocycle; R 1c [is H or -CH3] The compound or a pharmaceutically acceptable salt thereof is administered to the subject in combination with an immunomodulator.

[0026] In one embodiment, a compound of formula (Ia) is disclosed. In another embodiment, a pharmaceutically acceptable salt of the compound of formula (Ia) is disclosed.

[0027] In some embodiments of equation (Ia), R 1 is -H or -C(O)CH(R 1a )NH2;R 1a The alkyl group is selected from -H or -(C1~C6) alkyl groups.

[0028] In some embodiments of equation (Ia), R 1 is -H.

[0029] In some embodiments of equation (Ia), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a is -H and R 1b is -H.

[0030] In some embodiments of equation (Ia), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a is a -(C1~C6) alkyl group; R 1b is -H.

[0031] In some embodiments of equation (Ia), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a CH2OR 1c And; R 1b is -H.

[0032] In some embodiments of equation (Ia), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a and R 1b These, together with the atoms they bond to, form a five-membered heterocycle.

[0033] In any of the embodiments of formula (Ia) described above, the compound has the following structural formula: [ka] [In the formula, R 1 It is expressed by one of the following: [which is the same as defined above in equation (Ia)].

[0034] In one embodiment, formula (IIa) is disclosed: [ka] [wherein n is 0 or 1; R 2 [is a compound selected from -H or -(C1~C4) alkyl], or a pharmaceutically acceptable salt thereof.

[0035] In one embodiment, a compound of formula (IIa) is disclosed. In another embodiment, a pharmaceutically acceptable salt of the compound of formula (IIa) is disclosed.

[0036] In some embodiments of equation (IIa), R 3 is -H.

[0037] In some embodiments of equation (IIa), R3 It is a -(C1~C4) alkyl group.

[0038] In any of the embodiments of formula (IIa) described above, the compound has the following structural formula: [ka] [In the formula, R 2 It is expressed by one of the following: [which is the same as defined above in equation (IIa)].

[0039] In one embodiment, formula (Ib) [ka] [wherein n is 0 or 1; R 1 is -H or -C(O)CH(R 1a )NHR 1b and; R 1a These are -H, -(C1~C4) alkyl, and CH2OR 1c Selected from; R 1b is -H; or alternatively, R 1a and R 1b These, together with the atoms they bond to, form a five-membered heterocycle; R 1c [is H or -CH3] The compound or a pharmaceutically acceptable salt thereof is administered to the subject in combination with an immune checkpoint inhibitor.

[0040] In one embodiment, a compound of formula (Ib) is disclosed. In another embodiment, a pharmaceutically acceptable salt of the compound of formula (Ib) is disclosed.

[0041] In some embodiments of formula (Ib), R 1 is -H.

[0042] In some embodiments of formula (Ib), R 1 is -C(O)CH(R1a )NHR 1b And; R 1a is -H and R 1b is -H.

[0043] In some embodiments of formula (Ib), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a is a -(C1~C6) alkyl group; R 1b is -H.

[0044] In some embodiments of formula (Ib), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a CH2OR 1c And; R 1b is -H.

[0045] In some embodiments of formula (Ib), R 1 is -C(O)CH(R 1a )NHR 1b And; R 1a and R 1b These, together with the atoms they bond to, form a five-membered heterocycle.

[0046] In any of the above embodiments of formula (Ib), the compound has the following structural formula: [ka] [In the formula, R 1 It is expressed by one of the following: [in formula (Ib), it is the same as defined above].

[0047] In one embodiment, formula (IIb) is disclosed: [ka] [wherein n is 0 or 1; R 2[is a compound selected from -H or -(C1~C4) alkyl], or a pharmaceutically acceptable salt thereof.

[0048] In one embodiment, a compound of formula (IIb) is disclosed. In another embodiment, a pharmaceutically acceptable salt of the compound of formula (IIb) is disclosed.

[0049] In some embodiments of formula (IIb), R 2 is -H.

[0050] In some embodiments of formula (IIb), R 2 It is a -(C1~C4) alkyl group.

[0051] In some embodiments of formula (IIb), the compound has the following structural formula: [ka] [In the formula, R 2 It is expressed by one of the following: [which is the same as defined above in equation (IIb)].

[0052] In some embodiments, compounds of formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), and (IIa2) (including all their subgenera and species) are converted via intramolecular cyclization to compounds of formulas (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all their subgenera and species), and vice versa. That is, this is a reciprocal conversion process. Compounds of formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), and (IIa2) (including all their subgenera and species), and compounds of formulas (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all their subgenera and species), are each partially or completely converted to others depending on the conditions, such as the temperature, pressure, humidity, pH, and / or composition of the medium (e.g., solvent). This is illustrated in the scheme below: [ka] In the formula, R 1 This is the same as what is defined in equations (Ia) and (Ib) above.

[0053] In some embodiments, the compounds disclosed are those listed in Table 1, or pharmaceutically acceptable salts thereof.

[0054] [Table 1]

[0055] [Table 2]

[0056] [Table 3]

[0057] [Table 4]

[0058] [Table 5]

[0059] [Table 6]

[0060] The term "C1-C4 alkyl" includes acyclic alkyl moieties having 1 to 4 carbon atoms, while the term "C1-C6 alkyl" includes acyclic alkyl moieties having 1 to 6 carbon atoms. Examples of C1-C4 alkyl moieties include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, and tert-butyl.

[0061] The phrase “pharmaceutically acceptable salt” typically includes acid or base addition salts of formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all subgenera or species thereof), as well as acid or base addition salts that retain the biological efficacy and properties of the compounds in Table 1, which are not biologically undesirable or otherwise undesirable. In many cases, the compounds of formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all subgenera or species thereof), as well as the compounds in Table 1, can form acidic salts and / or basic salts in the presence of basic groups and / or carboxyl groups or similar groups.

[0062] Pharmacokinetically acceptable acid addition salts can be formed from inorganic and organic acids, such as acetate, aspartate, benzoate, besilate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphor sulfonate, chloride / hydrochloride, chlortheophyllonate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide / iodide, isethionate, lactate, and lactate. These may be tobionates, lauryl sulfates, malates, maleates, malons, mandelates, mesilates, methylsulfates, naphthoates, napsylates, nicotinates, nitrates, octadecanoates, oleates, oxalates, palmitates, palmoates, phosphates / hydrogen phosphates / dihydrogen phosphates, polygalacturonates, propions, stearates, succinates, subsalicylates, sulfates / hydrogen sulfates, tartrates, tosylates, and trifluoroacetates. Examples of inorganic acids from which salts can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of organic acids from which salts can be derived include acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, and sulfosalicylic acid.

[0063] Pharmaceutically acceptable base addition salts can be formed with inorganic bases and organic bases. Examples of inorganic bases from which the salt can be derived include ammonia and salts of ammonium, as well as metals in columns I - XII of the periodic table. In certain embodiments, the salt is derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly preferred salts include salts of ammonium, potassium, sodium, calcium, and magnesium. Examples of organic bases from which the salt can be derived include, for example, primary amines, secondary amines, and tertiary amines, substituted amines such as naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Specific organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0064] Pharmaceutically acceptable salts of the compounds of formula (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including any subgenera or species thereof), and the compounds of Table 1 can be synthesized from the basic or acidic moieties by conventional chemical methods. Generally, such salts are prepared by reacting the free acid form of these compounds with a stoichiometric amount of an appropriate base (e.g., Na + Ca 2+ Mg 2+ or K +It can be prepared by reacting with a hydroxide, carbonate, bicarbonate, or the like), or by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. Such reactions are typically carried out in water or an organic solvent, or in a mixture of both. Generally, when feasible, it is desirable to use a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. A list of further suitable salts can be found, for example, in “Remington’s Pharmaceutical Sciences,” 20th ed., Mack Publishing Company, Easton, Pa., (1985); Berge et al., “J. Pharm. Sci., 1977, 66, 1-19, and “Handbook of Pharmaceutical Salts: Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0065] Also, any formula given herein is intended to represent the formula (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including any subgenus or species thereof), as well as the unlabeled form and the isotope-labeled form of the compounds in Table 1. Isotope-labeled compounds have the structure represented by the formula shown herein, except that one or more atoms are replaced by atoms of the same element but with different mass numbers. Examples of isotopes that can be incorporated into the formula (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including any subgenus or species thereof), as well as the compounds in Table 1, and their pharmaceutically acceptable salts, include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, for example, 2 H, 3 H, 11 C, 13 C,14 C, 15 N, 35 S, 36 Cl, and 125 I is an example. Formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all subgenera or species thereof), and the isotope-labeled compounds of Table 1 may generally be prepared by conventional techniques known to those skilled in the art, or by processes similar to those described in the accompanying examples, using appropriate isotope-labeled reagents instead of the previously used unlabeled reagents.

[0066] Formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all subgenera or species thereof), and the compounds in Table 1, may have various isomeric forms. The terms “optical isomer,” “stereoisomer,” or “diastereoisomer” refer to any of the various stereoisomer configurations that may exist for formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all subgenera or species thereof), and the compounds in Table 1. Because substituents can be attached to the chiral centers of carbon atoms, it is understood that the compounds of this disclosure include enantiomers, diastereomers, and racemates. The term “enantiomer” includes pairs of stereoisomers that are mirror images of each other and cannot be superimposed. A 1:1 mixture of a pair of enantiomers is a racemic mixture. This term is used to indicate a racemic mixture as needed. The term “diastereomer” or “diastereoisomer” includes stereoisomers that have at least two chiral atoms but are not mirror images of each other. Absolute stereochemistry is indicated according to the Kahn-Ingold-Prelogue RS system. If a compound is a pure enantiomer, the stereochemistry at each chiral center may be indicated as either R or S. Divided compounds whose absolute configuration is unknown may be indicated as (+) or (-) depending on the direction in which they rotate plane polarization at the wavelength of the sodium D line (dextrorotatory or levorotatory). Formulas (Ia), (Ia1), (Ia2), (IIa), (IIa1), (IIa2), (Ib), (Ib1), (Ib2), (IIb), (IIb1), and (IIb2) (including all their subgenera or species), as well as certain compounds in Table 1, contain one or more chiral centers or axes, and thus may give rise to enantiomers, diastereomers, or other stereoisomers that can be defined as (R)- or (S)- from an absolute stereochemical standpoint. This disclosure is intended to include all such possible isomers (including racemic mixtures, optically pure forms, and intermediate mixtures).Optically active (R)-isomers and (S)-isomers can be prepared using chiral synthons or chiral reagents, or they can be separated using prior art known in the art (e.g., chiral HPLC).

[0067] Immunomodulators As used herein, “immunomodulatory agent” refers to an agent that enhances an immune response (e.g., an antitumor immune response). An immunomodulator may be an antibody or its antigen-binding fragment, a protein, a peptide, a DNA or RNA fragment, a small molecule, or a combination thereof. In some embodiments, an immunomodulator is an immune checkpoint inhibitor. In some embodiments, an immunomodulator is an immunostimulant. As used herein, “immune checkpoint inhibitor” means an agent (i.e., an immune checkpoint agent) that inhibits a protein or peptide that blocks the immune system from attacking, for example, cancer cells. In some embodiments, an immune checkpoint agent that blocks the immune system prevents the production and / or activation of T cells. In some embodiments, an immune checkpoint inhibitor is cytotoxic T lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD1), or programmed cell death ligand 1 (PD-L1). PD-L1 and PD1 form cell surface-bound ligand-receptor pairs that weaken the immune response and prevent an overreaction of the immune system in healthy individuals. In some embodiments, cancer cells hijack the normal PD-L1 / PD1 immune checkpoint mechanism by overexpressing ligand PD-L1 (which binds to PD1 on effector CD8 T cells), thereby preventing T cells from initiating an immune response against cancer cells and / or tumors. PD-L1 is frequently expressed in a wide variety of cancers. Overexpression of tumor PD-L1 correlates with poor prognosis in many cancers (see, e.g., Hamid et al., Expert Opin Biol Ther 13(6):847-861, 2013). As used herein, “immunostimulant” means a substance that stimulates the immune system by inducing or increasing the activation of any of its components, without having antigen specificity in the immune response. In some embodiments, the immunostimulant is a Toll-like receptor 3 (TLR3) agonist, such as polyinosinic acid:polycytidylic acid, also known as poly(I:C) or poly(I:C).

[0068] A review describing immune checkpoint pathways and their blockade by immune checkpoint inhibitors is provided by Pardoll in Nature Reviews Cancer (April, 2012), pages 252-264. Immune checkpoint inhibitors exert antitumor activity by blocking one or more endogenous immune checkpoint pathways that downregulate the antitumor immune response. Inhibition or blockade of immune checkpoint pathways typically involves inhibiting checkpoint receptor-ligand interactions by immune checkpoint inhibitors, thereby reducing or eliminating signaling and the resulting decrease in the antitumor response.

[0069] As used herein, “immune checkpoint inhibitor” means a drug (i.e., an immune checkpoint agent) that inhibits a protein or peptide that blocks the immune system from attacking, for example, cancer cells. An immune checkpoint inhibitor may be an antibody or its antigen-binding fragment, a protein, a peptide, a small molecule, or a combination thereof. In some embodiments, an immune checkpoint inhibitor that blocks the immune system prevents the production and / or activation of T cells. In some embodiments, an immune checkpoint agent is a cytotoxic T lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD1), programmed cell death ligand 1 (PD-L1), or an inhibitory receptor that recognizes HLA-E and is expressed by a subset of NK cells and T cells (such as NKG2A). PD-L1 and PD1 form cell surface-bound ligand-receptor pairs that weaken the immune response and prevent an overreaction of the immune system in healthy individuals. In some embodiments, cancer cells hijack the normal PD-L1 / PD1 immune checkpoint mechanism by overexpressing ligand PD-L1 (which binds to PD1 on effector CD8 T cells), thereby preventing T cells from initiating an immune response against cancer cells and / or tumors. PD-L1 is frequently expressed in a wide variety of cancers. Tumor PD-L1 overexpression correlates with poor prognosis in many cancers (see, e.g., Hamid et al., Expert Opin Biol Ther 13(6):847-861, 2013).

[0070] In some embodiments of this disclosure, immune checkpoint inhibitors inhibit signaling interactions between immune checkpoint receptors and their corresponding ligands. Immune checkpoint inhibitors may act by inhibiting (antagonizing) immune checkpoint receptors (some examples of receptors include CTLA-4, PD-1, and NKG2A) or by inhibiting ligands of immune checkpoint receptors (some examples of ligands include PD-L1 and PD-L2), thereby blocking the activation of the immune checkpoint pathway. In such embodiments, the effect of the immune checkpoint inhibitor is to reduce or eliminate downregulation of specific aspects of the immune system's antitumor response in the tumor microenvironment.

[0071] In some embodiments, immune checkpoint inhibitors inhibit the CTLA-4 pathway or the PD-L1 / PD1 pathway. In some embodiments, the immune checkpoint inhibitor is an antibody. In some embodiments, the immune checkpoint inhibitor comprises an antibody that inhibits CTLA-4, PD1, or PD-L1. Immune checkpoint inhibitors, immune checkpoint inhibitors and examples thereof are provided, for example, in International Publication No. 2016 / 062722.

[0072] In some embodiments, the immune checkpoint inhibitor is an anti-CTLA-4 antibody or its derivative or antigen-binding fragment. In embodiments, the anti-CTLA-4 antibody selectively binds to the CTLA-4 protein or its antigen-binding fragment. Examples of anti-CTLA-4 antibodies, as well as their derivatives and fragments, are described, for example, in U.S. Patent Nos. 6,682,736; 7,109,003; 7,123,281; 7,411,057; 7,807,797; 7,824,67; 8,143,379; 8,491,895; and U.S. Patent Application Publication 2007 / 0243184. In some embodiments, the anti-CTLA-4 antibody is tremelimumab or ipilimumab.

[0073] The immune checkpoint receptor cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) is expressed on T cells and is involved in signaling pathways that reduce T cell activation levels. CTLA-4 is thought to downregulate T cell activation through competitive binding and sequestration of CD80 and CD86. In addition, CTLA-4 is involved in T Reg It has been shown to be involved in enhancing the immunosuppressive activity of cells.

[0074] In some embodiments, the immune checkpoint inhibitor is an anti-PD-L1 antibody, a derivative thereof, or an antigen-binding fragment. In some embodiments, the anti-PD-L1 antibody, a derivative thereof, or an antigen-binding fragment selectively binds to the PD-L1 protein or a fragment thereof. Examples of anti-PD-L1 antibodies, as well as their derivatives and fragments, are described, for example, in International Publication Brochures 01 / 14556, 2007 / 005874, 2009 / 089149, 2011 / 066389, and 2012 / 145493; U.S. Patent Nos. 8,217,149 and 8,779,108; U.S. Patent Publication Nos. 2012 / 0039906, 2013 / 0034559, 2014 / 0044738, and 2014 / 0356353. In some embodiments, the anti-PD-L1 antibody is MEDI4736 (durvalumab), MDPL3280A, 2.7A4, AMP-814, MDX-1105, atezolizumab (MPDL3280A), or BMS-936559.

[0075] The immune checkpoint receptor programmed death 1 (PD-1) is expressed by activated T cells upon prolonged exposure to antigens. The association of PD-1 with its known binding ligands, PD-L1 and PD-L2, primarily occurs within the tumor microenvironment, leading to downregulation of the antitumor-specific T cell response. Both PD-L1 and PD-L2 are known to be expressed on tumor cells. PD-L1 and PD-L2 expression on tumors has been associated with reduced survival outcomes.

[0076] In some embodiments, the anti-PD-L1 antibody is MEDI4736, also known as durvalumab. In some embodiments, the anti-PD-L1 antibody comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identity to any of SEQ ID NOs: 1-8. MEDI4736 is an anti-PD-L1 antibody that is selective for the PD-L1 polypeptide and blocks the binding of PD-L1 to the PD-1 receptor and the CD80 receptor. MEDI4736 can mitigate the PD-L1-mediated suppression of human T cell activation in vitro and can further suppress tumor growth in xenograft models via a T cell-dependent mechanism. MEDI4736 is further described in U.S. Patent No. 8,779,108. The fragmentary crystalline (Fc) domain of MEDI4736 contains a triple mutation within the constant domain of the IgG1 heavy chain, which reduces its binding to complement components C1q and Fcγ receptors, which are responsible for mediating antibody-dependent cell-mediated cytotoxicity (ADCC).

[0077] In some embodiments, MEDI4736 or its antigen-binding fragment comprises a heavy chain and a light chain or a heavy chain variable region and a light chain variable region. In some embodiments, MEDI4736 or its antigen-binding fragment for use comprises a light chain variable region containing the amino acid sequence of SEQ ID NO: 1 and a heavy chain variable region containing the amino acid sequence of SEQ ID NO: 2. In some embodiments, MEDI4736 or its antigen-binding fragment comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 3-5, and the light chain variable region comprising the Kabat-defined CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 6-8. Those skilled in the art can readily identify the Chothia definition, the Abm definition, or other CDR definitions known to those skilled in the art. In some embodiments, MEDI4736 or its antigen-binding fragment comprises the variable heavy chain and variable light chain CDR sequences of the 2.14H90PT antibody described in International Publication Brochure 2011 / 066389.

[0078] In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody or its derivative or antigen-binding fragment. In some embodiments, the anti-PD-1 antibody selectively binds to the PD-1 protein or its antigen-binding fragment. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, or pidilizumab.

[0079] The NKG2A receptor is an inhibitory receptor that binds to HLA-E and is expressed in tumor-infiltrating cytotoxic NK and CD8 T lymphocytes. By expressing HLA-E, cancer cells can protect themselves from being killed by NKG2A+ immune cells. HLA-E is frequently upregulated in cancer cells of many solid tumors or hematological malignancies. Monalizumab (IPH2201), a humanized IgG4, blocks the binding of NKG2A to HLA-E, enabling activation of NK and cytotoxic T cell responses. Examples of anti-NKG2A antibodies and their derivatives and fragments are described in International Publication No. 2016 / 041947 (whose contents, including but not limited to sequence listings, are incorporated herein by reference in their entirety).

[0080] In some embodiments of this disclosure, the immune checkpoint inhibitor compound is an organic small molecule (molecular weight less than 1000 daltons), a peptide, a polypeptide, a protein, an antibody, an antibody fragment, or an antibody derivative. In some embodiments, the immune checkpoint inhibitor compound is an antibody. In some embodiments, the antibody is a monoclonal antibody, specifically a human or humanized monoclonal antibody.

[0081] Monoclonal antibodies, antibody fragments, and antibody derivatives for blocking immune checkpoint pathways can be prepared by any of several methods known to those skilled in the art, including, but not limited to, somatic cell hybridization techniques and hybridoma methods. The preparation of hybridomas is described in Antibodies, A Laboratory Manual, Harlow and Lane, 1988, Cold Spring Harbor Publications, New York. Human monoclonal antibodies can be identified and isolated by screening a phage display library of human immunoglobulin genes by methods described, for example, in U.S. Patent Nos. 5,223409, 5,403484, 5571698, 6582915, and 6593081. Monoclonal antibodies can also be prepared using the general method described in U.S. Patent No. 6,331415 (Cabilly).

[0082] For example, human monoclonal antibodies can be prepared using XenoMouse® (Abgenix, Freemont, CA) or hybridomas of XenoMouse-derived B cells. XenoMouse is a mouse host possessing functional human immunoglobulin genes as described in U.S. Patent No. 6,162,963 (Kucherlapati).

[0083] Methods for preparing and using immune checkpoint antibodies are described in the following exemplary publications: Preparation and therapeutic use of anti-CTLA-4 antibodies are described in U.S. Patent No. 7,229,628 (Allison), No. 7,311,910 (Linsley), and No. 8,017,144 (Korman). Preparation and therapeutic use of anti-PD-1 antibodies are described in U.S. Patent No. 8,008,449 (Korman) and U.S. Patent Application Publication No. 2011 / 0271358 (Freeman). Preparation and therapeutic use of anti-PD-L1 antibodies are described in U.S. Patent No. 7,943,743 (Korman). Preparation and therapeutic use of anti-TIM-3 antibodies are described in U.S. Patent No. 8,101,176 (Kuchroo) and No. 8,552,156 (Tagayanagi). The preparation and therapeutic use of anti-LAG-3 antibodies are described in U.S. Patent Publication No. 2011 / 0150892 (Thudium) and International Publication No. 2014 / 008218 (Lonberg). The preparation and therapeutic use of anti-KIR antibodies are described in U.S. Patent No. 8119775 (Moretta). The preparation of antibodies that block the BTLA regulatory inhibitory pathway (anti-BTLA antibodies) is described in U.S. Patent No. 8563694 (Mataraza).

[0084] In some embodiments of this disclosure, the immune checkpoint inhibitor compound is a CTLA-4 receptor inhibitor, a PD-1 receptor inhibitor, a LAG-3 receptor inhibitor, a TIM-3 receptor inhibitor, a BTLA receptor inhibitor, or a KIR receptor inhibitor. In some embodiments, the immune checkpoint inhibitor compound is a PD-L1 inhibitor or a PD-L2 inhibitor.

[0085] In some embodiments of this disclosure, the immune checkpoint inhibitor is an inhibitor of the PD-L1 / PD-1 pathway or the PD-L2 / PD-1 pathway. In some embodiments, the inhibitor of the PD-L1 / PD-1 pathway is MEDI4736.

[0086] In some embodiments of this disclosure, the immune checkpoint inhibitor compound is an anti-CTLA-4 receptor antibody, an anti-PD-1 receptor antibody, an anti-LAG-3 receptor antibody, an anti-TIM-3 receptor antibody, an anti-BTLA receptor antibody, an anti-KIR receptor antibody, an anti-PD-L1 antibody, or an anti-PD-L2 antibody.

[0087] In some embodiments of this disclosure, the anti-CTLA-4 receptor antibody is ipilimumab or tremelimumab. In some embodiments, the anti-PD-1 receptor antibody is lambrolizumab, pizilizumab, or nivolumab. In some embodiments, the anti-KIR receptor antibody is lirilumab.

[0088] Radiation therapy Radiotherapy, also known as high-dose ionizing radiation therapy, is used for cancer treatment in combination with compounds of formula (Ia) or formula (Ib), or pharmaceutically acceptable salts thereof. In some embodiments, the method involves administering radiotherapy to a patient in combination with compounds of formula (Ia) or formula (Ib), or pharmaceutically acceptable salts thereof, and an immunomodulator.

[0089] Radiotherapy may involve X-rays, gamma rays, or charged particles. Radiotherapy may also be external beam radiotherapy or internal radiotherapy (also known as proximal radiotherapy). Whole-body radiotherapy using radioactive materials such as radioactive iodine may also be used. Examples of external beam radiotherapy include 3D structured radiotherapy, intensity-controlled radiotherapy, image-guided radiotherapy, tomotherapy, stereotactic radiotherapy, proton therapy, or other charged particle beams.

[0090] Radiotherapy may involve X-rays, gamma rays, or charged particles. Radiotherapy may also be external beam radiotherapy or internal radiotherapy (also known as proximal radiotherapy). Whole-body radiotherapy using radioactive materials such as radioactive iodine may also be used. Examples of external beam radiotherapy include 3D structured radiotherapy, intensity-controlled radiotherapy, image-guided radiotherapy, tomotherapy, stereotactic radiotherapy, proton therapy, or other charged particle beams.

[0091] In some embodiments, the radiotherapy is fractionated radiotherapy. In one embodiment, the fractionated radiotherapy comprises 2 to 14 fractions. In another embodiment, the fractionated radiotherapy comprises 2 to 7 fractions. In yet another embodiment, the fractionated radiotherapy comprises 3 to 6 fractions. In one embodiment, the fractionated radiotherapy comprises 2, 3, 4, 5, 6, or 7 fractions. In one embodiment, the fractionated radiotherapy comprises 5 fractions. In some embodiments, the radiotherapy fractions are administered daily. In one embodiment, the radiotherapy may comprise two or more daily doses and / or daily doses. In one configuration, the radiotherapy fractions are administered on days 1, 2, 3, 4, and 5.

[0092] Combination therapy This disclosure provides a method for treating cancer, wherein a compound of formula (Ia) or (Ib) (including any subgenus or species thereof), or a pharmaceutically acceptable salt thereof, and an immunomodulator are administered in combination. In another embodiment, this disclosure provides a method for treating cancer, wherein a compound of formula (Ia) or (Ib) (including any subgenus or species thereof), or a pharmaceutically acceptable salt thereof, and radiotherapy are administered in combination, optionally together with an immunomodulator.

[0093] In some embodiments, the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and the immunomodulator are administered sequentially, separately, or simultaneously. In some embodiments, the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and radiotherapy are administered on the same day or on different days. In one embodiment, radiotherapy is used before treatment with the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and / or the immunomodulator. In another embodiment, radiotherapy is used after treatment with the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and / or the immunomodulator. In yet another embodiment, radiotherapy is administered simultaneously with treatment with the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and / or the immunomodulator.

[0094] In some embodiments, the compound of formula (Ia) or (Ib) is selected from the compounds listed in Table 1, i.e., compounds 1 to 33, and the immunomodulator is selected from durvalumab, tremelimumab, monalizumab, or a combination thereof.

[0095] In some embodiments, administration of a compound of formula (Ia) or (Ib) in combination with an immunomodulator results in additive and / or synergistic effects. As used herein, the term “synergistic” refers to a combination of therapies that is more effective than the additive effect of a single therapy (e.g., MEDI4736 or this antigen-binding fragment in combination with an arginase inhibitor described herein).

[0096] In some embodiments, the administration of a combination of the compound of formula (Ia) or (Ib) and an immunomodulatory agent by the method provided herein, for example, favorably enhances antigen presentation and / or promotes T cell activation, thereby providing a safer and more effective treatment for the patient compared to the administration of a single agent alone. In some embodiments, the method provided herein results in an increase in CD8+ T cells, NK cells, and / or CD103+ dendritic cells compared to the administration of an immunomodulatory agent alone or a compound of formula (Ia) or (Ib) alone. In some embodiments, the method provided herein results in an increase in the patient's interferon-γ (IFNγ) levels compared to the administration of a single agent alone. In some embodiments, the method provided herein results in an increase in the patient's interleukin-2 (IL-2) levels compared to the administration of a single agent alone.

[0097] Sequence IDs 1-8 correspond to the amino acid sequences of MEDI4736, an anti-PD-L1 antibody as described in the embodiments herein. Sequence ID 3 corresponds to the amino acid sequence of the light chain variable region of MEDI4736. Sequence ID 4 corresponds to the amino acid sequence of the heavy chain variable region of MEDI4736. Sequence IDs 5-10 correspond to the CDR of MEDI4736.

[0098] Sequence IDs 9-16 correspond to the amino acid sequences of tremelimumab, an anti-CTLA-4 antibody as described in the embodiments herein.

[0099] Sequence IDs 17-24 correspond to the amino acid sequences of monalizumab, an anti-NKG2A antibody as described in the embodiments herein.

[0100] All references cited herein, such as patents, patent applications, articles, textbooks, etc., and references cited therein, are incorporated herein by reference in their entirety, unless they have already been cited. [Examples]

[0101] The aspects of this disclosure may be further defined by reference to the non-limiting examples described below, which describe in detail the preparation of certain compounds and intermediates of the disclosure, as well as methods of using the compounds of the disclosure. It will be apparent to those skilled in the art that many modifications to both materials and methods can be made without departing from the scope of this disclosure.

[0102] Unless otherwise specified, (i) Unless otherwise specified, all synthesis shall be carried out at ambient temperature (i.e., within the range of 17-25°C) and in an atmosphere of an inert gas such as nitrogen; (ii) Evaporation is carried out by rotary evaporation or in a vacuum using a Genevac apparatus or Biotage v10 evaporator, and after removing residual solids by filtration, post-treatment procedures are performed; (iii) Flash chromatography purification was performed using a pre-packed RediSep Rf Gold® silica column (20-40 μm, spherical particles), a GraceResolv® cartridge (Davisil® silica), or a Silicycle cartridge (40-63 μm) on an automated Teledyne Isco CombiFlash® Rf or Teledyne Isco CombiFlash® Companion® system. (iv) Preparative chromatography was performed using a Gilson preparative HPLC instrument equipped with UV collection; or preparative chromatography was performed using a Waters AutoPurification HPLC-MS instrument equipped with MS and UV trigger collection; (v) Chiral preparative chromatography was performed using a Gilson instrument with UV collection (233 injector / fraction collector, 333 & 334 pumps, 155 UV detector) or a Varian Prep Star instrument (2×SD1 pump, 325 UV detector, 701 fraction collector) pump (running with Gilson 305 injection); or chiral preparative chromatography was performed using a Waters Prep 100 SFC-MS instrument with MS and UV trigger collection, or a Thar MultiGram III SFC instrument with UV collection. (vi) The yield, if any, is not necessarily the maximum achievable value; (vii) The structure of the final product of equation I was generally confirmed by nuclear magnetic resonance (NMR) spectroscopy; NMR chemical shift values ​​were measured on a delta scale [proton magnetic resonance spectra were obtained using a Bruker Avance III 600 (600 MHz), Bruker Avance 400 (400 MHz), Bruker Avance 300 (300 MHz), or Bruker DRX 500 (500 MHz) instrument]; measurements were performed at ambient temperature unless otherwise specified; the following abbreviations were used: s, singlet; d, doublet; t, triplet; q, quadruplet; m, multiplet; dd, doublet of doublet; ddd, doublet of doublet of doublet; dt, triplet of doublet; bs, broad signal. (viii) Generally, the final product of formula I is also characterized by mass spectrometry after liquid chromatography (LCMS or UPLC); UPLC is performed using a Waters UPLC equipped with a Waters SQ mass spectrometer (column temperature 40°C, UV = 220-300 nm or 190-400 nm, mass spectrometry = ESI with positive / negative switching) at a flow rate of 1 mL / min for 1.50 minutes (total runtime to return to the starting state etc. is 1.70 minutes) using a solvent system of 97%A + 3%B to 3%A + 97%B, where A = 0.1% formic acid or 0.05% trifluoroacetic acid in water (for acidic treatment), or 0.1% ammonium hydroxide in water (for basic treatment), and B = acetonitrile. The column used for acid analysis was Waters Acquity HSS T3 (1.8 μm, 2.1 × 50 mm), and the column used for basic analysis was Waters Acquity BEH C18 (1.7 μm, 2.1 × 50 mm). Alternatively, UPLC was performed using a Waters UPLC equipped with a Waters SQ mass spectrometer (column temperature 30°C, UV = 210-400 nm, mass spectrometry = ESI with positive / negative switching) at a flow rate of 1 mL / min for 1.5 minutes (total runtime to return to equilibrium to the starting state was 2 minutes) using a solvent gradient of 2-98% B, where A = 0.1% formic acid in water and B = 0.1% formic acid in acetonitrile (for acid treatment), or A = 0.1% ammonium hydroxide in water and B = acetonitrile (for basic treatment).The column used for acid analysis was Waters Acquity HSS T3 (1.8 μm, 2.1 × 30 mm), and the column used for basic analysis was Waters Acquity BEH C18 (1.7 μm, 2.1 × 30 mm); LCMS was performed over 4 minutes with a flow rate of 1.1 mL / min using a Waters Alliance HT (2795) and a Phenomenex Gemini-NX C18 (5 μm, 110A, 2.1 × 50 mm) column, with a retention of 0.5 minutes at 95% A to 95% B, where A = 0.1% formic acid in acetonitrile and B = 0.1% formic acid (for acid treatment), or A = 0.1% ammonium hydroxide in water and B = acetonitrile (for basic treatment). In addition, Shimadzu UFLC and Waters HSS were used with Shimadzu LCMS-2020 mass spectrometers. Using a C18 (1.8μm, 2.1×50mm), Shim-pack XR-ODS (2.2μm, 3.0×50mm), or Phenomenex Gemini-NX C18 (3μm, 3.0×50mm) column, the flow rate is 0.7mL / min (for Waters HSS C18 column), 1.0mL / min (for Shim-pack XR-ODS column), or 1.2mL / min (for Phenomenex Gemini-NX column). LCMS was performed for 2.2 minutes at the flow rate (for C18), with a 0.6 minute hold at 95%A to 95%B, where A = 0.1% formic acid or 0.05% trifluoroacetic acid in water (for acidic treatment), or 0.1% ammonium hydroxide or 6.5 mM ammonium carbonate in water (for basic treatment), and B = acetonitrile. Reported molecular ions correspond to [M+H]+ unless otherwise specified; for molecules with multiple isotopic patterns (Br, Cl, etc.), the reported values ​​are obtained at the lowest isotopic mass unless otherwise specified. (ix) Ion exchange purification was generally performed using the SCX-2 (Biotage) cartridge. (x) The purity of the intermediate is evaluated by thin-layer chromatography, mass spectrometry, LC-MS, UPLC / MS, HPLC (high-performance liquid chromatography), and / or NMR analysis; (xi) The following abbreviations were used: EtOH: Ethanol æ:ethyl acetate LDA: Lithium diisopropylamide MeOH: methanol TFA: Trifluoroacetic acid MeCN: Acetonitrile LCMS: Liquid Chromatography Mass Spectrometry rt or RT: Room temperature aq: water-based THF: Tetrahydrofuran KHMDS: Potassium bis(trimethylsilyl)amide DCM: Dichloromethane DMF: Dimethylformamide HATU: (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate) BOC: tert-butoxycarbonyl DTNB: 5,5'-Dithiobis(2-nitrobenzoic acid) TNB: 2-Nitro-5-thiobenzoic acid HEPES: (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid).

[0103] The preparations of compounds 1-9 in Table 1 are shown in Examples 1-9 below. Furthermore, the preparations of compounds 10-33 in Table 1 are described as Examples 7-30 in International Publication No. 2019 / 159120, and their contents are incorporated in their entirety by reference.

[0104] Example 1: (2R,4S)-4-amino-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0105] Intermediate 1: 2-benzyl 1-(tert-butyl)(R)-4-oxopiperidine-1,2-diccarboxylate N,N'-diisopropylcarbodiimide (3.49 mL, 22.4 mmol) and DMAP (0.249 g, 2.04 mmol) were added at 0°C to a stirred solution of (R)-1-(tert-butoxycarbonyl)-4-oxopiperidine-2-carboxylic acid (4.955 g, 20.37 mmol) and benzyl alcohol (2.11 mL, 20.4 mmol) in DCM (150 mL). The reaction mixture was stirred for 17 hours while slowly warming to room temperature. The reaction mixture was filtered, and the filtrate was concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-50% ethyl hexane) to obtain 2-benzyl 1-(tert-butyl)(R)-4-oxopiperidine-1,2-dicarboxylate (intermediate 1, 6.50 g, yield 96%) as a colorless oil and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 1.40(5H,br s),1.49(4H,br s),2.43-2.65(2H,m),2.70-2.92(2H,m),3.53-3.74(1H,m),3.94-4.10(1H,m),4.89(0.5H,br s),5.10-5.23(2.5H,m),7.31-7.41(5H,m);m / z:(ES + )[M+Na] + =356.

[0106] Intermediate 2: 2-benzyl 1-(tert-butyl)(2R,4R)-4-hydroxypiperidine-1,2-dicarboxylate Sodium borohydride (0.738 g, 19.5 mmol) was added in small amounts at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(R)-4-oxopiperidine-1,2-dicarboxylate (intermediate 1, 6.504 g, 19.51 mmol) in a MeOH / THF (1:20, 105 mL) mixture. The mixture was stirred for 5 hours, then carefully quenched with 1 M HCl (aq) (15 mL, gas generation) and warmed to room temperature. The mixture was diluted with water (25 mL) and ethyl acetate (50 mL). The phases were separated, and the aqueous phase was extracted with ethyl acetate (4 × 20 mL). The combined organic matter was washed with saturated NaCl aqueous solution, dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-45% SiO2 in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4R)-4-hydroxypiperidine-1,2-dicarboxylate (intermediate 2, 3.84 g, yield 59%) as a colorless rubber mixture of 5:1 diastereomers (the larger diastereomer being the title compound) and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 1.38(5H,br s),1.40-1.44(2H,m),1.46(5.4H,br s),1.60-1.69(1.4H,m),1.70-1.79(0.2H,m),1.83-1.97(1.2H,m),2.48(1H,br dd),2.92-3.08(1H,m),3.28-3.46(0.2H,m),3.54-3.70(1H,m),3.76-3.84(0.1H,m),3.87-3.96(0.1H,m),4.00(0.5H,br m / z:(ES + )[M+Na] + =358.

[0107] Intermediate 3: 2-benzyl 1-(tert-butyl)(2R,4R)-4-((methylsulfonyl)oxy)piperidine-1,2-dicarboxylate Methanesulfonic anhydride (3.59 g, 20.6 mmol) was added in small amounts at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4R)-4-hydroxypiperidine-1,2-dicarboxylate (intermediate 2, 3.84 g, 11.5 mmol; a mixture of 5:1 diastereomers) and triethylamine (3.35 mL, 24.0 mmol) in DCM (50 mL) at 0°C. The cooling bath was terminated, and the reaction mixture was warmed to room temperature. After 6 hours, the reaction mixture was diluted in DCM (50 mL) and washed sequentially with 1 M HCl (aq) (50 mL) and saturated NaCl aqueous solution (50 mL). The organic layer was dried over MgSO4, filtered, concentrated to dryness, and crude 2-benzyl 1-(tert-butyl)(2R,4R)-4-((methylsulfonyl)oxy)piperidine-1,2-dicarboxylate (intermediate 3, 4.73 g, yield 100%) was obtained as a pale orange rubber and as a mixture of diastereomers. The crude material was used directly without further purification. m / z:(ES + )[M+Na] + = 436.

[0108] Intermediate 4: 2-benzyl 1-(tert-butyl)(2R,4S)-4-azidopiperidine-1,2-dicarboxylate Sodium azide (3.72 g, 57.2 mmol) was added to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4R)-4-((methylsulfonyl)oxy)piperidine-1,2-dicarboxylate (intermediate 3, 4.73 g, 11.4 mmol; mixture of diastereomers) in DMF (50 mL), and the reaction was heated to 60°C for 20 hours. The mixture was cooled to room temperature and filtered, and the filtrate was diluted with water (400 mL) and SiO (40 mL). The phases were separated, and the aqueous phase was extracted with SiO (4 × 40 mL). The combined organic matter was washed with saturated NaCl aqueous solution (2 × 40 mL), dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-30% ethyl hexane) to obtain the pure diastereomer 2-benzyl 1-(tert-butyl)(2R,4S)-4-azidopiperidine-1,2-dicarboxylate (intermediate 4, 2.58 g, yield 63%) as a colorless rubber and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 1.39(4H,br s),1.46(5H,br s),1.63-1.73(1H,m),1.74-1.87(1H,m),1.95(1H,ddd),2.50-2.61(1H,m),3.03-3.44 (1H,m),3.73-3.89(0.5H,m),3.90-4.01(1.5H,m),4.58-4.74(0.5H,m),4.88(0.5H,br s),5.15-5.34(2H,m),7.30-7.43(5H,m);m / z:(ES + )[M-Boc] + =261.

[0109] Intermediate 5: 2-benzyl 1-(tert-butyl)(4S)-4-azido-2-(buto-2-en-1-yl)piperidine-1,2-dicarboxylate 2-Benzyl 1-(tert-butyl)(2R,4S)-4-azidopiperidine-1,2-dicarboxylate (intermediate 4, 1.94 g, 5.38 mmol) and clotyl bromide (0.977 mL, 8.07 mmol) were dissolved in THF (30 mL), and the solution was cooled to -78°C. KHMDS solution (1 M in 2-methyltetrahydrofuran, 7.0 mL, 7.0 mmol) was added dropwise over 10 minutes. The reaction mixture was slowly warmed to room temperature and stirred for a total of 18 hours. The crude reaction mixture was quenched with saturated NH4Cl aqueous solution, then diluted with saturated NaCl aqueous solution and SiO2 (50 mL). The phases were separated, and the aqueous layer was extracted with SiO2 (3 × 30 mL). The combined organic matter was washed with saturated NaCl aqueous solution, dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (2-30% ethyl hexane) to obtain 2-benzyl 1-(tert-butyl)(4S)-4-azido-2-(buto-2-en-1-yl)piperidine-1,2-dicarboxylate (intermediate 5, 2.106 g, yield 94%) as a syn / antidiastereomer mixture and as a mixture of rotational isomers and E / Z olefins. 1 H NMR(500MHz,CD2Cl2)δ 1.40-1.42(4H,m),1.43(5H,br s),1.49-1.58(1H,m),1.59-1.66(0.6H,m),1.67-1.74(3.4H,m),1.86(0.5H,dd),1.89-2.05(2H,m),2.07 -2.19(1H,m),2.42(0.5H,dd),2.58-2.69(1H,m),2.71-2.83(0.5H,m),3.01-3.16(0.5H,m),3.21(0.5H,br dd),3.31-3.44(0.5H,m),3.61-3.77(1.5H,m),3.97-4.07(0.5H,m),5.10-5 .27(2H,m),5.36-5.45(1H,m),5.51-5.74(2H,m),7.32-7.47(5H,m);m / z:(ES + )[M-Boc] + =315.

[0110] Intermediate 6: 2-benzyl 1-(tert-butyl)(2R,4S)-4-azide-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate Bis(1,5-cyclooctadiene)diiridium(I) dichloride (50 mg, 0.074 mmol) and bis(diphenylphosphino)methane (57 mg, 0.15 mmol) were added to an oven-dried round-bottom flask. The flask was sealed and purged with N2. The solids were dissolved in DCM (9 mL), and 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.32 mL, 2.2 mmol) was slowly added to the solution. The reaction mixture was stirred at room temperature for 10 minutes. 2-benzyl 1-(tert-butyl)(4S)-4-azido-2-(buto-2-en-1-yl)piperidine-1,2-dicarboxylate (intermediate 5, 616 mg, 1.49 mmol) was added to the reaction mixture as a solution in DCM (3 mL), and the reaction mixture was stirred at room temperature for 66 hours. The reaction mixture was cooled to 0°C and carefully quenched with MeOH (1 mL) and water (5 mL). The layers were separated, and the aqueous layer was extracted with DCM (3 times × 15 mL). The combined organic matter was dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-15% ethyl hexane) to obtain the pure diastereomer 2-benzyl 1-(tert-butyl)(2R,4S)-4-azido-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 6, 261 mg, yield 32%) as a clear, colorless rubber. 1H NMR(500MHz,CDCl3)δ 0.79(2H,t),1.25(12H,s),1.29-1.35(1H,m),1.36-1.39(1H,m),1.4 1(9H,s),1.42-1.46(2H,m),1.57-1.68(1H,m),1.85-1.94(3H,m),1.9 5-2.01(1H,m),2.05(1H,dd),2.92-3.11(1H,m),3.49-3.72(1H,m),3. 98-4.03(1H,m),5.09(1H,d),5.18(1H,d),7.29-7.42(5H,m);m / z:(ES + )[M+Na] + = 565.

[0111] Intermediate 7: (2R,4S)-4-amino-1-(tert-butoxycarbonyl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 50 mg, 0.047 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-azido-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 6, 268 mg, 0.494 mmol) in ELISA (3 mL). The suspension was stirred at room temperature for 17 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with MeOH (5 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-45% MeOH in DCM) to obtain (2R,4S)-4-amino-1-(tert-butoxycarbonyl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 7, 156 mg, yield 74%) as a white dry film. 1H NMR(500MHz,CD2Cl2)δ 0.71(2H,t),1.07-1.16(1H,m),1.19(14H,s),1.31-1.37(2H,m),1.40(9H,s),1.80-1.96(1H,m),2.02(3H,br d),2.33(1H,br s),3.00(1H,br s),3.53(1H,br s),3.92(1H,br s),8.60(3H,br s);m / z:(ES + )[M+H] + = 427.

[0112] Example 1: (2R,4S)-4-amino-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.53 mL, 6.9 mmol) was added dropwise to a stirred solution of (2R,4S)-4-amino-1-(tert-butoxycarbonyl)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 7, 146 mg, 0.342 mmol) in DCM (2 mL) at room temperature. After 2 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (3.0 mL, 3.0 mmol) and Et2O (3 mL). Phenylboronic acid (125 mg, 1.03 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (20 mL) and water (5 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20cc column). The desired product was eluted from the column with 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-amino-2-(4-boronobutyl)piperidine-2-carboxylic acid (62 mg, yield 74%) as a white solid. 1H NMR(500MHz,D2O)δ 0.71-0.82(2H,m),1.10-1.30(2H,m),1.33-1.44(2H,m),1.45-1.55(1H,m),1.62(1H,dd),1.77(1H,ddd),1 m / z:(ES + )[M+H] + =245.

[0113] Example 2: (2R,4S)-4-((S)-2-amino-3-methylbutanamide)-2-(4-voronobutyl)piperidine-2-carboxylic acid [ka]

[0114] Intermediate 8: 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate Zinc (270 mg, 4.14 mmol) and AcOH (1.20 mL, 20.9 mmol) were added to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-azido-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 6, 748 mg, 1.38 mmol) in THF (10 mL). The rapidly stirred mixture was heated at 30°C for 18 hours. The mixture was cooled to room temperature, diluted with DCM (30 mL), and filtered through diatomaceous earth. The filter cake was washed with DCM, and the filtrate was concentrated to dryness. The resulting residue was partitioned between siRNA (40 mL) and saturated NaHCO3 aqueous solution. The phases were separated, and the organic matter was washed with saturated NaHCO3 aqueous solution and saturated NaCl aqueous solution. The organic matter was dried over MgSO4, filtered, and concentrated to dryness to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 713 mg, yield 100%) as a clear, colorless rubber. The crude material was used directly without further purification. 1 H NMR(500MHz,CDCl3)δ 0.79(2H,t),1.24(12H,s),1.32-1.38(2H,m),1.39-1.47(13H,m),1.68(2H,br m / z:(ES + )[M+H] + = 517.

[0115] Intermediate 9: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.12 mL, 0.63 mmol) was slowly added at room temperature to a stirred solution of COMU (270 mg, 0.63 mmol) and Boc-Val-OH (137 mg, 0.631 mmol) in DMF (2 mL). The solution was stirred at room temperature for 30 minutes and then cooled to 0°C. 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 310 mg, 0.60 mmol) in DMF (2 mL) and N,N-diisopropylethylamine (0.10 mL, 0.60 mmol) was added, and the reaction mixture was stirred for 17 hours while slowly warming it to room temperature. The reaction mixture was diluted with water (40 mL), and the resulting precipitate was collected by filtration. The solid was dissolved in ethyl acetate, dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (10-100% ethyl acetate in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 9, 184 mg, yield 43%) as a colorless film and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.73-0.80(2H,m),0.85(3H,d),0.89(3H,d),1.22(12H,br s),1.23-1.29(2H,m),1.37-1.45(20H,m),1.62-1.74(1H,m),1.86-1.99(3H,m),2.00-2 .12(3H,m),2.97(1H,t),3.78(1H,t),3.94-4.06(1H,m),4.07-4.14(1H,m),5.00(1H,br s),5.05-5.24(2H,m),6.05(1H,br d), 7.28-7.36 (5H, m); m / z: (ES + )[M+H] + =716.

[0116] Intermediate 10: (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 27 mg, 0.025 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 9, 184 mg, 0.257 mmol) in ELISA (2 mL). The suspension was stirred at room temperature for 16 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (20 mL) and MeOH (20 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (20-100% ethyl hexane followed by 10% MeOH in DCM) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 10, 116 mg, yield 72%) as a white solid and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.78(3H,br d),0.83-0.91(2H,m),0.94(3H,br d),1.20-1.25(12H,m),1.40(9H,br s),1.42-1.53(11H,m),1.51-1.66(1H,m),1.75-2.18(4H,m),2.19-2.34(1 H,m),2.88-3.06(1H,m),3.85-4.06(2H,m),4.07-4.26(1H,m),5.14(1H,br s),5.93(1H,br s),6.73(1H,br s),7.30-7.48(1H,m);m / z:(ES+ )[M+H] + =627.

[0117] Example 2: (2R,4S)-4-((S)-2-amino-3-methylbutanamide)-2-(4-voronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.433 mL, 5.63 mmol) was added dropwise to a stirred solution of (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 10, 176 mg, 0.281 mmol) in DCM (2 mL) at room temperature. After 3 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (3.0 mL, 3.0 mmol) and Et2O (3 mL). Phenylboronic acid (103 mg, 0.845 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (20 mL) and water (5 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion exchange chromatography (PoraPak Rxn CX 20 cc column). The desired product was eluted from the column with 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-((S)-2-amino-3-methylbutanamide)-2-(4-voronobutyl)piperidine-2-carboxylic acid (Example 2, 89 mg, yield 92%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.73-0.83(2H,m),0.88-0.96(6H,m),1.14-1.24(1H,m),1.25-1.35(1H,m),1.37-1.50(2H,m),1.64-1.76(1H,m),1.79- m / z:(ES + )[M+H] + =344.

[0118] Example 3: (2R,4S)-4-(2-aminoacetamide)-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0119] Intermediate 11: 2-benzyl 1-(tert-butyl)(2R,4S)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.12 mL, 0.63 mmol) was slowly added at room temperature to a stirred solution of COMU (270 mg, 0.63 mmol) and Boc-Gly-OH (110 mg, 0.63 mmol) in DMF (2 mL). The solution was stirred at room temperature for 30 minutes and then cooled to 0°C. A solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 310 mg, 0.60 mmol) in DMF (2 mL) and N,N-diisopropylethylamine (0.11 mL, 0.60 mmol) was added, and the reaction mixture was stirred for 17 hours while slowly warming it to room temperature. The reaction mixture was diluted with water (60 mL) and the pH was adjusted to approximately 5 with acetic acid. The aqueous phase was extracted with ethyl acetate (4 × 15 mL). The combined organic matter was washed with saturated aqueous NaCl solution (2 × 10 mL), dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (10-100% ethyl acetate in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 11, 204 mg, yield 51%) as a colorless film and as a mixture of rotational isomers. 1H NMR(500MHz,CDCl3)δ 0.75(2H,br t),1.21(14H,s),1.24(2H,br d),1.38(9H,s),1.40(10H,s),1.71(1H,dd),1.81-1.91(1H,m),1.93-2.04(3H,m),2.86-3.04(1H,m),3.62(2H,br m / z:(ES + )[M+H] + =674.

[0120] Intermediate 12: (2R,4S)-1-(tert-butoxycarbonyl)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 32 mg, 0.030 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 11, 204 mg, 0.303 mmol) in ELISA (2 mL). The suspension was stirred at room temperature for 16 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (20 mL) and MeOH (20 mL), filtered through diatomaceous earth, and the filtrate was concentrated to dryness. The resulting residue was purified by flash silica chromatography (25-100% ethyl acetate in hexane) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 12, 117 mg, yield 66%) as a white solid and as a mixture of rotational isomers. 1H NMR(500MHz,CDCl3)δ 0.78(2H,t),1.17-1.29(13H,m),1.40(10H,br s),1.45(9H,s),1.48-1.57(2H,m),1.76-2.01(3H,m),2.04-2.13(1H,m),2.98(1H,br t),3.47-3.66(1H,m),3.75(1H,s),3.90-4.06(2H,m),4.13-4.25(1H,m),5.41(1H,br s),5.92(1H,br s),6.73(1H,br s),7.65(1H,br s);m / z:(ES + )[M+H] + = 584.

[0121] Example 3: (2R,4S)-4-(2-aminoacetamide)-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.31 mL, 4.0 mmol) was added dropwise to a stirred solution of (2R,4S)-1-(tert-butoxycarbonyl)-4-(2-((tert-butoxycarbonyl)amino)acetamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 12, 117 mg, 0.201 mmol) in DCM (2 mL) at room temperature. After 3 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (3.0 mL, 3.0 mmol) and Et2O (3 mL). Phenylboronic acid (73 mg, 0.60 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (20 mL) and water (5 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20cc column). The desired product was eluted from the column with 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-(2-aminoacetamide)-2-(4-voronobutyl)piperidine-2-carboxylic acid (Example 3, 61 mg, yield 100%) as a white solid. 1H NMR(500MHz,D2O)δ 0.72-0.84(2H,m),1.14-1.25(1H,m),1.26-1.34(1H,m),1.41(2H,quin),1.72(1H,dtd),1.79-1.94(2H,m),1 .96-2.08(2H,m),2.10(1H,dd),3.14-3.25(1H,m),3.30-3.36(1H,m),3.37(2H,s),4.08-4.19(1H,m);m / z:(ES + )[M+H] + =302.

[0122] Example 4: (2R,4S)-4-[[(2S)-2-aminopropanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0123] Intermediate 13: 2-benzyl 1-tert-butyl(2R,4S)-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.17 mL, 1.0 mmol) was slowly added at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 245 mg, 0.474 mmol), Boc-Ala-OH (108 mg, 0.571 mmol), and COMU (244 mg, 0.571 mmol) in DMF (1.5 mL). The reaction mixture was stirred for 1.5 hours while slowly warming to room temperature. The reaction mixture was diluted with water (20 mL) and ethyl acetate (20 mL), and the phases were separated. The aqueous phase was extracted with SiO2 (3 × 20 mL), the combined organic matter was washed with saturated NaCl aqueous solution, dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (15-80% SiO2 in hexane) to obtain 2-benzyl 1-tert-butyl(2R,4S)-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-1,2-dicarboxylate (intermediate 13, 191 mg, yield 59%) as a pale yellow rubber and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.69-0.83(2H,m),1.22(12H,d),1.26(5H,td),1.38(9H,br s),1.40(9H,br d),1.42-1.49(3H,m),1.57-1.73(1H,m),1.83-1.98(3H,m),2.01-2.05(2H,m),2.91-3.03(1H,m),4.02(2H,br s),4.96(1H,br s),5.05-5.22(2H,m),6.21(1H,br s),7.27-7.36(5H,m);m / z:(ES + )[M+H] + =688.

[0124] Intermediate 14: (2R,4S)-1-tert-butoxycarbonyl-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-2-carboxylic acid Pd / C (10 wt%, 15 mg, 0.014 mmol) was added to a solution of 2-benzyl 1-tert-butyl (2R,4S)-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-1,2-dicarboxylate (intermediate 13, 190 mg, 0.28 mmol) in ELISA (2 mL). The suspension was stirred at room temperature for 17 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (15 mL) and MeOH (2 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (20-100% ethyl hexane) to obtain (2R,4S)-1-tert-butoxycarbonyl-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-2-carboxylic acid (intermediate 14, 134 mg, yield 81%) as a dry film and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.78(2H,br t),1.20-1.25(12H,m),1.29-1.36(5H,m),1.41(9H,s),1.44(11H,s),1.48-1.62(2H,m),1.77-2.01(3H,m),2.09(2H,br s),2.97(1H,br m / z:(ES),3.92-4.05(1H,m),5.07(1H,br s),5.48(1H,br s),6.71(1H,br s),7.61(1H,br s); + )[M+H] + = 598.

[0125] Example 4: (2R,4S)-4-[[(2S)-2-aminopropanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.34 mL, 4.4 mmol) was added dropwise to a stirred solution of (2R,4S)-1-tert-butoxycarbonyl-4-[[(2S)-2-(tert-butoxycarbonylamino)propanoyl]amino]-2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl]piperidine-2-carboxylic acid (intermediate 14, 130 mg, 0.22 mmol) in DCM (1 mL) at room temperature. After 3 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (2.0 mL, 2.0 mmol) and Et2O (2 mL). Phenylboronic acid (80 mg, 0.65 mmol) was added, and the clear two-phase solution was stirred at room temperature for 3 hours. The mixture was diluted with Et2O (20 mL) and water (5 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20cc column). The desired product was eluted from the column using 5% ammonia in MeOH (20 mL). The obtained substance was further purified by reverse-phase chromatography (RediSep Rf Gold® C18, 0-15% acetonitrile in water) to obtain (2R,4S)-4-[[(2S)-2-aminopropanoyl]amino]-2-(4-voronobutyl)piperidine-2-carboxylic acid (Example 4, 25 mg, yield 37%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.72-0.80(2H,m),1.12-1.23(1H,m),1.24-1.26(1H,m),1.27(3H,d),1.40(2H,quin),1.66-1.76(1H,m),1.78-1.92(2H,m) m / z:(ES + )[M+H] + =316.

[0126] Example 5: (2R,4S)-4-[[(2S)-2-aminobutanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0127] Intermediate 15: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.165 mL, 0.94 mmol) was slowly added at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 244 mg, 0.47 mmol), Boc-Abu-OH (96 mg, 0.47 mmol), and COMU (206 mg, 0.48 mmol) in DMF (3 mL). The reaction mixture was stirred for 16 hours while slowly warming to room temperature. The crude reaction mixture was diluted with water (30 mL) and extracted with ELISA (3 × 10 mL). The combined organic matter was sequentially washed with saturated NaHCO3 aqueous solution (20 mL) and saturated NaCl aqueous solution (15 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (15-60% ethyl hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 15, 215 mg, yield 65%) as a colorless rubber and as a mixture of rotational isomers. 1H NMR(500MHz,CDCl3)δ 0.71-0.79(2H,m),0.87(3H,br t),1.19(4H,br s),1.21(9H,s),1.36(5H,br s),1.38(8H,s),1.39-1.41(8H,m),1.48-1.58(2H,m),1.68(1H,br dd),1.72-1.81(1H,m),1.84-1.98(3H,m),1.99-2.02(1H,m),2.88-3.04(1H,m),3.89(1H,br d),3.95-4.07(2H,m),5.00(1H,br d),5.05-5.22(2H,m),6.20(1H,br s),7.27-7.36(5H,m);m / z:(ES + )[M+H] + =703.

[0128] Intermediate 16: (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10% wt, 16 mg, 0.015 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 15, 215 mg, 0.31 mmol) in ELISA (3 mL). The suspension was stirred at room temperature for 24 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (10 mL) and MeOH (1 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-100% ethyl acetate in hexane) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 16, 147 mg, yield 78%) as a dry film and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.75(2H,br s),0.89(3H,br s),1.20(12H,s),1.24-1.33(2H,m),1.37(10H,br s),1.40(10H,s),1.42-1.61(4H,m),1.85(3H,br s),1.98(2H,br s),2.01-2.06(1H,m),2.95(1H,br s),3.98(2H,br s),5.27(0.5H,br s),5.68(0.5H,br s),6.74(0.5H,br s),7.52(0.5H,br s);m / z:(ES + )[M+H] + =612.

[0129] Example 5: (2R,4S)-4-[[(2S)-2-aminobutanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.37 mL, 4.8 mmol) was added dropwise to a stirred solution of (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)butanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 16, 147 mg, 0.24 mmol) in DCM (1 mL) at room temperature. After 2 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (2.0 mL, 2.0 mmol) and Et2O (2 mL). Phenylboronic acid (88 mg, 0.72 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (5 mL) and water (2 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20cc column). The desired product was eluted from the column using 5% ammonia in MeOH (20 mL). The obtained substance was further purified by reverse-phase chromatography (RediSep Rf Gold® C18, 0-15% acetonitrile in water) to obtain (2R,4S)-4-[[(2S)-2-aminobutanoyl]amino]-2-(4-voronobutyl)piperidine-2-carboxylic acid (Example 5, 37 mg, yield 47%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.76(2H,br t),0.87(3H,t),1.13-1.23(1H,m),1.24-1.34(1H,m),1.40(2H,quin),1.63(2H,dq),1 .67-1.74(1H,m),1.78-1.85(1H,m),1.86-1.96(2H,m),1.99-2.07(1H,m),2.13(1H,br dd),3.10-3.24(1H,m),3.25-3.41(2H,m),4.07-4.21(1H,m);m / z:(ES + )[M+H] + =330.

[0130] Example 6: (2R,4S)-4-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0131] Intermediate 17: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.17 mL, 0.94 mmol) was added at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 244 mg, 0.47 mmol), Boc-Leu-OH (96 mg, 0.47 mmol), and COMU (206 mg, 0.48 mmol) in DMF (3 mL). The reaction mixture was stirred for 16 hours while slowly warming to room temperature. The crude reaction mixture was diluted with water (30 mL) and extracted with siRNA (3 × 10 mL). The combined organic matter was sequentially washed with saturated NaHCO3 aqueous solution (20 mL) and saturated NaCl aqueous solution (15 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (15-60% ethyl hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 17, 224 mg, yield 65%) as a white foam and as a mixture of rotational isomers. 1H NMR(500MHz,CDCl3)δ 0.73-0.80(2H,m),0.89(3H,d),0.90(3H,d),1.22(12H,s),1.27(1H,br s),1.39(9H,s),1.40(9H,s),1.41-1.48(4H,m),1.55-1.64(2H,m),1.68(1H,br dd),1.85-1.98(3H,m),2.01(1H,br d),2.02-2.05(1H,m),2.93-3.02(1H,m),3.94-4.08(3H,m),4.83(1H,br d),5.05-5.22(2H,m),6.20(1H,br s),7.28-7.31(1H,m),7.33(4H,d);m / z:(ES + )[M+H] + =731.

[0132] Intermediate 18: (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 13 mg, 0.012 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 17, 174 mg, 0.24 mmol) in ELISA (2.5 mL). The suspension was stirred at room temperature for 23 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (10 mL) and MeOH (1 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (15-100% ethyl hexane) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 18, 149 mg, yield 98%) as a dry film and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.78(2H,br t),0.91(6H,br d),1.23(11H,s),1.27-1.36(2H,m),1.40(10H,s),1.43(11H,s),1.46-1.55(2H,m),1.60-1.73(2H,m),1. 78-1.96(3H,m),1.97-2.02(1H,m),2.05-2.14(1H,m),2.82-3.08(1H,m),3.92-4.08(2H,m),5.00(0.4H,br s),5.49(0.6H,br d),6.76(0.4H,br d),7.60(0.6H,br s);m / z:(ES + )[M+H] + =640.

[0133] Example 6: (2R,4S)-4-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.36 mL, 4.7 mmol) was added dropwise to a stirred solution of (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-4-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 18, 149 mg, 0.23 mmol) in DCM (1 mL) at room temperature. After 2 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (2.0 mL, 2.0 mmol) and Et2O (2 mL). Phenylboronic acid (85 mg, 0.70 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (10 mL) and water (2 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20 cc column). The desired product was eluted from the column with 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-[[(2S)-2-amino-4-methyl-pentanoyl]amino]-2-(4-voronobutyl)piperidine-2-carboxylic acid (Example 6, 81 mg, yield 97%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.74-0.82(2H,m),0.89(3H,d),0.91(3H,d),1.15-1.25(1H,m),1.24- 1.34(1H,m),1.38-1.47(3H,m),1.47-1.54(1H,m),1.60(1H,dt),1.65 -1.76(1H,m),1.79-1.98(3H,m),2.01-2.09(1H,m),2.14(1H,dd),3.15-3.23(1H,m),3.31(1H,dt),3.39(1H,t),4.10-4.17(1H,m);m / z:(ES + )[M-H2O+H] + =340.

[0134] Example 7: (2R,4S)-4-[[(2S,3S)-2-amino-3-methyl-pentanoyl]amino]-2-(4-voronobutyl)piperidine-2-carboxylic acid [ka]

[0135] Intermediate 19: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.24 mL, 1.4 mmol) was slowly added at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 355 mg, 0.687 mmol), Boc-Ile-OH (159 mg, 0.687 mmol), and COMU (300 mg, 0.70 mmol) in DMF (4 mL). The reaction mixture was stirred for 16 hours while slowly warming to room temperature. The crude reaction mixture was diluted with water (30 mL), and the mixture was stirred for 10 minutes. The resulting precipitate was collected by filtration. The solid was dissolved in ELISA (20 mL), washed with saturated NaCl aqueous solution (5 mL), dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-60% ELISA in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 19, 424 mg, yield 85%) as a white foamy substance and as a mixture of rotational isomers. 1H NMR(500MHz,CDCl3)δ 0.73-0.80(2H,m),0.82-0.90(6H,m),0.97-1.11(1H,m),1.21(12H,s),1.26-1.31(1H,m),1.32-1.36(1H,m),1.38(9H,br s),1.39(9H,s),1.41-1.43(2H,m),1.67(1H,br dd),1.75-1.84(2H,m),1.86-1.98(3H,m),2.00(1H,br d),2.03(1H,br m / z:(ES + )[M+H] + =730.

[0136] Intermediate 20: (2R,4S)-1-(tert-butoxycarbonyl)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 22 mg, 0.021 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 19, 302 mg, 0.41 mmol) in ELISA (4 mL). The suspension was stirred at room temperature for 20 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (10 mL) and MeOH (1 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (15-100% ethyl acetate in hexane, followed by 0-50% MeOH in ethyl acetate) and (2R,4S)-1-(tert-butoxycarbonyl)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-cal Boronic acid (intermediate 20, 223 mg, yield 84%) was obtained as a dry film and as a mixture of rotational isomers, and the boronic acid byproduct (2R,4S)-2-(4-boronobutyl)-1-(tert-butoxycarbonyl)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)piperidine-2-carboxylic acid (30 mg, yield 13%) was obtained as a white solid and as a mixture or rotational isomer. Intermediate 20: 1H NMR(500MHz,CDCl3)δ 0.72-0.81(2H,m),0.82-0.88(3H,m),0.89-0.94(3H,m),0.98-1.11(1H,m ),1.21-1.24(12H,m),1.26-1.31(1H,m),1.32-1.38(2H,m),1.40(10H,br s),1.42(10H,br s),1.45-1.55(1H,m),1.53-1.64(1H,m),1.76-1.97(4H,m),1.98-2.08(3H,m),2.83-3.10(1H,m),3.90-4.08(2H,m),5.15(0.5H,br d),5.81(0.5H,br s),6.74(0.5H,br s),7.48(0.5H,br s);m / z:(ES + [M+H] + =640. By-products of poromonic acid: 1 H NMR(500MHz, CDCl3)δ 0.76-0.86(2H,m),0.86-0.96(6H,m),1.03-1.13(1H,m),1.43(9H,br s),1.44(12H,s),1.53-1.68(1H,m),1.78-1.90(2H,m),1.90-1.99(2H,m),1.99- 2.04(1H,m),2.07-2.21(1H,m),2.98-3.11(1H,m),3.43-3.61(1H,m),3.90(1H,br s),3.96-4.12(2H,m),5.19(1H,br s),5.53-5.76(1H,m),6.72(1H,br s);m / z:(ES + [M+H] + =558.

[0137] Example 7: (2R,4S)-4-[[(2S,3S)-2-アミノ-3-メチル-ペンタノイル]アミノ]-2-(4-ボロノブチル)ピペリジン-2-カルボン acid Trifluoroacetic acid (0.62 mL, 8.1 mmol) was added dropwise to a stirred solution in DCM (2 mL) containing (2R,4S)-1-(tert-butoxycarbonyl)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 20 mg, 223 mg, 0.35 mmol) and the boronic acid by-product (2R,4S)-2-(4-boronobutyl)-1-(tert-butoxycarbonyl)-4-((2S,3S)-2-((tert-butoxycarbonyl)amino)-3-methylpentanamide)piperidine-2-carboxylic acid (30 mg, 0.05 mmol) at room temperature. After 2 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (3.0 mL, 3.0 mmol) and Et2O (3 mL). Phenylboronic acid (147 mg, 1.21 mmol) was added, and the clear two-phase solution was stirred at room temperature for 3 hours. The mixture was diluted with Et2O (5 mL) and water (1 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20 cc column). The desired product was eluted from the column using 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-[[(2S,3S)-2-amino-3-methyl-pentanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid (Example 7, 126 mg, yield 88%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.78(2H,td),0.85-0.91(6H,m),1.11-1.25(2H,m),1.26-1.34(1H,m),1.37-1.49(3H,m),1.64-1.76(2H,m),1.80-1.91 (2H,m),1.92-1.99(1H,m),2.01-2.09(1H,m),2.17(1H,dd),3.15-3.24(2H,m),3.31(1H,dt),4.10-4.21(1H,m);m / z:(ES + )[M+H] + =358.

[0138] Example 8: (2R,4S)-4-[[(2S)-2-amino-3,3-dimethyl-butanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0139] Intermediate 21: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.24 mL, 1.4 mmol) was slowly added at 0°C to a stirred solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 355 mg, 0.687 mmol), Boc-tert-Leu-OH (159 mg, 0.687 mmol), and COMU (300 mg, 0.70 mmol) in DMF (4 mL). The reaction mixture was stirred for 16 hours while slowly warming to room temperature. The crude reaction mixture was diluted with water (30 mL), and the mixture was stirred for 10 minutes. The resulting precipitate was collected by filtration. The solid was dissolved in SiO2 (20 mL), washed with saturated NaCl aqueous solution (5 mL), dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-55% SiO2 in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 21, 372 mg, yield 74%) as a white foam. 1H NMR(500MHz,CDCl3)δ 0.73-0.81(2H,m),0.95(9H,s),1.24(12H,s),1.41(10H,s),1.42-1.46(12H,m),1.68(1H,d d),1.87-1.95(1H,m),1.96-2.01(2H,m),2.02-2.05(2H,m),2.92-3.04(1H,m),3.69(1H,br d),3.98-4.12(2H,m),5.05-5.25(3H,m),5.50-5.62(1H,m),7.30-7.39(5H,m);m / z:(ES + )[M+H] + =730.

[0140] Intermediate 22: (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 27 mg, 0.025 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 21, 372 mg, 0.511 mmol) in ELISA (4 mL). The suspension was stirred at room temperature for 20 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (10 mL) and MeOH (1 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (40-100% ethyl acetate in hexane, followed by 0-40% MeOH in ethyl acetate) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-ca Rubonic acid (intermediate 22, 265 mg, yield 81%) was obtained as a dry film and a mixture of rotational isomers, and the boronic acid byproduct (2R,4S)-2-(4-boronobutyl)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)piperidine-2-carboxylic acid (32 mg, yield 11%) was obtained as a transparent dry film and a mixture of rotational isomers. Intermediate 22: 1H NMR(500MHz, CDCl3)δ 0.75(2H,br t),0.93(9H,s),1.20(12H,s),1.24-1.33(2H,m),1.36(9H,s),1.38(9H,br s),1.40(3H,br s),1.51-1.64(1H,m),1.72-1.82(1H,m),1.83-1.98(3H,m),2.03(1H,br s),2.79-3.06(1H,m),3.76(0.4H,br d),3.85-4.07(2.6H,m),5.42(0.6H,br d),6.39-6.71(1H,m),6.74-6.99(0.4H,m);m / z:(ES + [M+H] + =640. By-products of poromonic acid: 1 H NMR(500MHz, CDCl3)δ 0.77-0.86(2H,m),0.97(9H,s),1.42(9H,br s),1.43(14H,br s),1.52-1.67(1H,m),1.75-1.90(2H,m),1.91-2.04(3H,m),2.08-2.19(1H,m),2.97-3.14(1H,m),3.44-3.63(1H,m),3.87(1H,br d),3.97-4.11(2H,m),5.50(1H,br s),6.53-6.79(1H,m);m / z:(ES + [M+H] + =558.

[0141] Example 8: (2R,4S)-4-[[(2S)-2-アミノ-3,3-ジメチル-ブタノイル]アミノ]-2-(4-ボロノブチル)ピペリジン-2-カルボン acid Trifluoroacetic acid (0.73 mL, 9.4 mmol) was added dropwise at room temperature to a stirred solution in DCM (2 mL) of (2R,4S)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 22 mg, 265 mg, 0.414 mmol) and the boronic acid by-product (2R,4S)-2-(4-boronobutyl)-1-(tert-butoxycarbonyl)-4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethylbutanamide)piperidine-2-carboxylic acid (32 mg, 0.057 mmol). After 2 hours, the solution was concentrated under reduced pressure, and the resulting residue was dissolved in 1 M HCl (aq) (3.0 mL, 3.0 mmol) and Et2O (3 mL). Phenylboronic acid (172 mg, 1.41 mmol) was added, and the clear two-phase solution was stirred at room temperature for 4 hours. The mixture was diluted with Et2O (5 mL) and water (1 mL), and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was freeze-dried and purified by ion-exchange chromatography (PoraPak Rxn CX 20 cc column). The desired product was eluted from the column using 5% ammonia in MeOH (20 mL) to obtain (2R,4S)-4-[[(2S)-2-amino-3,3-dimethyl-butanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid (Example 8, 158 mg, yield 94%) as a white solid. 1 H NMR(500MHz,D2O)δ 0.74-0.81(2H,m),0.95(9H,s),1.15-1.25(1H,m),1.26-1.34(1H,m),1.38-1.48(2H,m),1.65-1.75(1H,m),1.80-1.91(2H,m ),1.96(1H,ddd),2.01-2.10(1H,m),2.19(1H,dd),3.05(1H,s),3.15-3.24(1H,m),3.30(1H,dt),4.10-4.23(1H,m);m / z:(ES + )[M+H] + =358.

[0142] Example 9: (2R,4S)-4-[[(2R)-2-amino-3-methyl-butanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid [ka]

[0143] Intermediate 23: 2-benzyl 1-(tert-butyl)(2R,4S)-4-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate N,N-diisopropylethylamine (0.063 mL, 0.36 mmol) was slowly added at 0°C to a stirred solution of HATU (61 mg, 0.16 mmol) and Boc-D-Val-OH (33 mg, 0.15 mmol) in DMF (1 mL). The solution was stirred for 10 minutes, and then a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-amino-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 8, 75 mg, 0.15 mmol) in DMF (1 mL) was added. The reaction mixture was stirred for 16 hours while slowly warming it to room temperature. The crude reaction mixture was diluted with 0.1 M HCl (aq) (30 mL) and ethyl acetate. The phases were separated, and the aqueous phase was extracted with SiO2 (3 × 15 mL). The combined organic matter was washed with saturated NaCl aqueous solution, dried over MgSO4, filtered, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (5-40% SiO2 in hexane) to obtain 2-benzyl 1-(tert-butyl)(2R,4S)-4-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 23, 74 mg, yield 71%) as a colorless film and as a mixture of rotational isomers. 11H NMR (500 MHz, CDCl3) δ 0.75 (2H, t), 0.84 (3H, d), 0.89 (3H, d), 1.21 (12H, s), 1.30 - 1.35 (1H, m), 1.37 (9H, s), 1.39 (9H, s), 1.42 (4H, br s), 1.69 (1H, dd), 1.81 - 1.91 (1H, m), 1.92 - 2.01 (3H, m), 2.02 (2H, s), 2.94 - 3.02 (1H, m), 3.75 (1H, dd), 3.91 - 4.03 (1H, m), 5.03 - 5.19 (3H, m), 6.10 (1H, d), 7.28 - 7.39 (5H, m); m / z: (ES + ) [M + H] + = 715.

[0144] Intermediate 24: (2R,4S)-1-(tert-Butoxycarbonyl)-4-((R)-2-((tert-Butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid Pd / C (10 wt%, 5 mg, 0.005 mmol) was added to a solution of 2-benzyl 1-(tert-butyl)(2R,4S)-4-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-1,2-dicarboxylate (intermediate 23, 69 mg, 0.10 mmol) in ELISA (1 mL). The suspension was stirred at room temperature for 19 hours under a hydrogen atmosphere (the balloon flask was evacuated and refilled with hydrogen three times). The reaction mixture was diluted with ELISA (10 mL) and MeOH (1 mL), filtered through diatomaceous earth, and concentrated to dryness. The resulting residue was purified by flash silica chromatography (40-100% ethyl acetate in hexane, followed by 0-40% MeOH in ethyl acetate) to obtain (2R,4S)-1-(tert-butoxycarbonyl)-4-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (intermediate 24, 40 mg, yield 66%) as a dry film and as a mixture of rotational isomers. 1 H NMR(500MHz,CDCl3)δ 0.77(2H,t),0.91(6H,br d),1.20-1.24(13H,m),1.29(1H,br d),1.33-1.42(11H,m),1.43(10H,s),1.70-1.82(1H,m),1.83-1.90(1H,m),1. 91-2.02(4H,m),3.00(1H,t),3.83-3.94(1H,m),3.96-4.07(1H,m),4.13(1H,br s),5.40(1H,d),6.03-6.33(1H,m),6.90-7.22(1H,m);m / z:(ES + )[M+H] + = 625.

[0145] Example 9: (2R,4S)-4-[[(2R)-2-amino-3-methyl-butanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid Trifluoroacetic acid (0.10 mL, 1.3 mmol) was added dropwise at room temperature to a stirred solution of (2R,4S)-1-(tert-butoxycarbonyl)-4-((R)-2-((tert-butoxycarbonyl)amino)-3-methylbutanamide)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)piperidine-2-carboxylic acid (Intermediate 24, 40 mg, 0.06 mmol) in DCM (1 mL). After 2 hours, the solution was concentrated under reduced pressure and the resulting residue was dissolved in 1 M HCl(aq) (1.0 mL, 1.0 mmol) and Et2O (1 mL). Phenylboronic acid (24 mg, 0.20 mmol) was added and the clear biphasic solution was stirred at room temperature for 20 hours. The mixture was diluted with Et2O (5 mL) and water (1 mL) and the layers were separated. The aqueous layer was washed with Et2O. The aqueous layer was lyophilized and purified by ion exchange chromatography (PoraPak Rxn CX 20 cc column). The desired product was eluted from the column using 5% ammonia in MeOH (20 mL) to give (SR,4S)-4-[[(2R)-2-amino-3-methyl-butanoyl]amino]-2-(4-boronobutyl)piperidine-2-carboxylic acid (Example 9, 20 mg, 91% yield) as a white solid. 1 H NMR(500MHz,D2O)δ 0.73-0.83(2H,m),0.87-0.95(6H,m),1.14-1.23(1H,m),1.24-1.33(1H,m),1.41(2H,quin),1.65-1.77(1H,m),1.79-1.87(1H,m),1.88-1.98(3H,m),2.01-2.09(1H,m),2.15(1H,dd),3.14(1H,d),3.15-3.24(1H,m),3.31(1H,dt),4.11-4.20(1H,m);m / z:(ES + )[M-H2O+H] + =326.

[0146] Example 10. Efficacy of Compound 12, an arginase 1 inhibitor, alone or in combination with anti-PDL1 or anti-PDL1 and anti-NKG2a or poly I:C in a preclinical model of cancer Method: MC38-OVA: Mouse MC38 colorectal cancer cells expressing the OVA antigen (5 × 10⁻¹⁰ 5 Cells (mouse) were subcutaneously transplanted into the right flank of 6-8 week old female C57BL / 6 mice. Six days post-transplantation, groups of mice were treated with a vehicle (water), 30 mg / kg of compound 12, 10 mg / kg of anti-PDL1 (MEDI4736), or a combination thereof. Compound 12 was administered orally twice daily in water. MEDI4736 was administered intraperitoneally twice weekly for two weeks in 1XPBS. The length and width of the tumors were measured using calipers, and the formula (length × width) was used. 2 ) * The tumor volume was calculated using π / 6 and then reported as the calculated tumor volume.

[0147] To monitor changes in immune cells, tumors were harvested at either 4, 10, or 14 days post-treatment. The tumors were excised and mechanically chopped. The tumors were then incubated with a tumor dissociation enzyme mix (Miltenyi Biotec) in a mild MACS instrument (Miltenyi Biotec) at 37°C for 40 minutes. Single-cell suspensions were prepared and stained for various immunocellular markers to measure changes in immune cells post-treatment using multicolor flow cytometry. Furthermore, the functional response of CD8+ cytotoxic T cells was evaluated by ex vivo restimulation with phorbol 12-myristate-13-acetate (PMA) and ionomycin prior to analysis.

[0148] CT.26 WT: Mouse CT.26 WT colorectal cancer cells (5 × 10 5Cells (mouse) were subcutaneously transplanted into the right flank of 6-8 week old female Balb / C mice. On day 6 post-transplantation, one group of mice was treated with a vehicle (water), 30 mg / kg of compound 12, 10 mg / kg of anti-PDL1 (MEDI4736), or a combination thereof. An additional group was treated with 10 mg / kg of anti-NKG2A (monalizumab) in combination with either compound 12 or MEDI4736, or with a tripartite combination of compound 12, monalizumab, and MEDI4736. Compound 12 was administered orally twice daily in water. MEDI4736 was administered intraperitoneally twice weekly for 2 weeks in 1XPBS. Monalizumab was administered intravenously twice weekly for 1.5 weeks in 1XPBS (3 doses). The length and width of the tumor were measured using calipers, and the formula (length × width) was used. 2 ) * The tumor volume was calculated using π / 6 and then reported as the calculated tumor volume.

[0149] LL / 2: Mouse Lewis lung (LL / 2) cancer cells (1 × 10⁻¹⁰ 6 Cells (from a mouse) were subcutaneously transplanted into the right flank of 6-8 week old C57BL / 6 mice. Six days post-transplant, the mouse groups were treated with a vehicle (water), 30 mg / kg of compound 12, 7.5 mg / kg of a TLR3 agonist (PolyI:C), or a combination thereof. Compound 12 was administered orally twice daily in water. PolyI:C was administered intraperitoneally three times a week in water. The length and width of the tumors were measured using calipers, and the formula (length × width) was used. 2 ) * The tumor volume was calculated using π / 6 and then reported as the calculated tumor volume.

[0150] Results: As shown in Figures 1A-1D, 2A-2D, and 3A-3F, monotherapy with compound 12 was moderately effective in all three mouse syngenic tumor models. Anti-PDL1 administered alone also showed moderate efficacy in both colorectal cancer tumor models. Combinations of drugs showed significantly stronger efficacy: 87% tumor growth inhibition in MC38-OVA and 46% overall tumor growth inhibition with one complete remission in the CT.26 WT model. Addition of anti-NKG2a to compound 12 + anti-PDL1 treatment resulted in 53% overall tumor growth inhibition and three complete remissions. The TLR3 agonist, poly-I:C, showed moderate monotherapy activity at TGI on day 16, at 76%. The combination of compound 12 + poly-I:C resulted in 87% tumor growth inhibition.

[0151] As shown in Figures 4A-4F, in vivo PD combination studies of ARG inhibitors with anti-PDL1 in the MC38-OVA model showed an increase in multiple tumor immune cell populations (approximately 4x CD8+ T cells, 2x NK cells, 2x CD103+ DCs) and increased CD8 T cell activation. Furthermore, as shown in Figures 5A and 5B, the ARG inhibitors resulted in an increase in surrounding IFNg and TNFa-producing CD8+ T cells (effluxing LNs) and showed inhibition with one complete remission in the CT.26WT model.

[0152] Example 11. Efficacy of compound 12, an arginase-1 inhibitor, in combination with radiotherapy (RT) in a preclinical model of cancer. Methods: 6-8 week old BL / 6 mice (Charles River Labs) were transplanted subcutaneously into the right flank with 1e6 LL / 2 mouse lung cancer cells. Six days post-transplant, the animals were randomized into four groups (n=12 / group) based on mean cage tumor volume. Group 1 mice were anesthetized for simulated irradiation on days 6 and 9, and started with vehicle (water) PO and BID on day 6. Group 2 mice were anesthetized and irradiated with 5 Gy of targeted radiation on days 6 and 9. Group 3 mice were started with AZ8400 at 30 mg / kg PO-BID on day 6, and Group 4 mice were started with AZ8400 at 30 mg / kg on day 6, and anesthetized and irradiated on days 6 and 9. Body weight and tumor volume were considered for mice with tumors >1500 mm². 3 The mice were measured twice a week until they reached a certain point, at which point they were humanely slaughtered.

[0153] result: As shown in Figures 5A and 5B, the combination of compound 12 and radiotherapy demonstrated remarkably potent efficacy in the mouse syngenic lung tumor model LL / 2, achieving approximately 60% tumor growth inhibition compared to administration of compound 12 at 30 mg / kg twice daily with radiotherapy (5 Gray on days 6 and 9 post-transplant; approximately 30% tumor growth inhibition, p>0.05 compared to vehicle) (p<0.05 compared to vehicle and monotherapy).

[0154] [Table 7]

[0155] [Table 8]

[0156] [Table 9]

Claims

1. A method for treating cancer patients, using formula (Ia) or (Ib): 【Chemistry 1】 [wherein n is 0 or 1; R １ is -H or -C(O)CH(R １ａ ) NHR １ｂ And; R １ａ is -H, -(C １ ~C ６ ) alkyl, and CH ２ OR １ｃ Selected from; R １ｂ is -H; or alternatively, R １ａ and R １ｂ together with the atoms to which they are attached form a 5-membered heterocycle; R １ｃ is H or -CH ３ [is] A method comprising administering to the patient an effective amount of the compound or a pharmaceutically acceptable salt thereof, and an effective amount of an immunomodulator.

2. R １ is -H or -C(O)CH(R １ａ ) NH ２ And; R １ａ is -H or -(C １ ~C ６ ) Selected from alkyl groups, The method according to claim 1.

3. The compound of formula (Ia) or (Ib) is formula (IIa) or (IIb): 【Chemistry 2】 [wherein n is 0 or 1; R ２ is -H or -(C １ ~C ４ ) Selected from alkyl groups] The method according to claim 1 or 2, as represented by the following:

4. The method according to any one of claims 1 to 3, wherein the compound of formula (Ia) or (Ib) is selected from the compounds in Table 1.

5. The method according to any one of claims 1 to 4, wherein the immunomodulator is an immune checkpoint inhibitor or an immunostimulator.

6. The method according to claim 5, wherein the immune checkpoint inhibitor is selected from a CTLA-4 receptor inhibitor, a PD-1 receptor inhibitor, a PD-L1 inhibitor, a PD-L2 inhibitor, an NKG2A receptor inhibitor, or a combination thereof.

7. The method according to claim 5, wherein the immunostimulant is a TLR3 agonist.

8. The method according to any one of claims 1 to 6, wherein the immune checkpoint inhibitor is an antibody or an antigen-binding fragment thereof.

9. The method according to any one of claims 1 to 6 and 8, wherein the immune checkpoint inhibitor is an anti-CTLA-4 receptor antibody, an anti-PD-1 receptor antibody, an anti-PD-L1 antibody, an anti-PD-L2 antibody, or an anti-NKG2A receptor antibody.

10. The method according to claim 9, wherein the immune checkpoint inhibitor is durvalumab, tremelimumab, monalizumab, or a combination thereof.

11. The method according to any one of claims 1 to 10, wherein the cancer is breast cancer, bladder cancer, head and neck cancer, non-small cell lung cancer, small cell lung cancer, colorectal cancer, gastrointestinal stromal tumor, gastroesophageal cancer, renal cell carcinoma, prostate cancer, liver cancer, colon cancer, pancreatic cancer, ovarian cancer, melanoma, non-Hodgkin lymphoma, cutaneous T-cell lymphoma, multiple myeloma, acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), chronic myeloid leukemia (CML), chronic lymphocytic leukemia (CLL), chronic myelomonocytic leukemia (CMML), and diffuse large B-cell lymphoma (DLBCL).

12. The method according to any one of claims 1 to 11, wherein the compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, is administered sequentially, separately, or simultaneously with the immunomodulator.

13. A compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer patients, which is administered to the patient sequentially, separately, or simultaneously with an immunomodulator.

14. An immunomodulator for use in the treatment of cancer, wherein an immune checkpoint inhibitor is administered to the patient sequentially, separately, or simultaneously with a compound of formula (Ia) or (Ib), or a pharmaceutically acceptable salt thereof.

15. i) Equation (Ia) or (Ib): 【Transformation 3】 [wherein n is 0 or 1; R １ is -H or -C(O)CH(R １ａ ) NHR １ｂ And; R １ａ is -H, -(C １ ~C ６ ) alkyl, and CH ２ OR １ｃ Selected from; R １ｂ is -H; or alternatively, R １ａ and R １ｂ These, together with the atoms they bond to, form a five-membered heterocycle; R １ｃ is H or -CH ３ [is] ii) Pharmaceutical products comprising the compounds or pharmaceutically acceptable salts thereof, and immunomodulators.

16. The pharmaceutical product according to claim 15, wherein the compound of (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and the immunomodulator are available in a single dosage form.

17. The pharmaceutical product according to claim 15, wherein the compound of (Ia) or (Ib), or a pharmaceutically acceptable salt thereof, and the immunomodulator are present in a separation agent form.

18. A method for treating cancer patients, using formula (Ia) or (Ib): 【Chemistry 4】 [wherein n is 0 or 1; R １ is -H or -C(O)CH(R １ａ ) NHR １ｂ And; R １ａ is -H, -(C １ ~C ６ ) alkyl, and CH ２ OR １ｃ Selected from; R １ｂ is -H; or alternatively, R １ａ and R １ｂ These, together with the atoms they bond to, form a five-membered heterocycle; R １ｃ is H or -CH ３ [is] A method comprising administering to the patient an effective amount of the compound or a pharmaceutically acceptable salt thereof, and an effective amount of radiotherapy.

19. The method according to claim 18, wherein the compound (Ia) or (Ib) is the same as that defined in any one of claims 2 to 4.

20. The method according to claim 18 or 19, wherein the radiotherapy is fractionated radiotherapy.

21. The method according to any one of claims 18 to 20, further comprising administering an effective amount of an immunomodulator to the patient.

22. The method according to any one of claims 18 to 21, wherein the immunomodulator is the same as that defined in any one of claims 5 to 10.

Images