Use of anti-aging glycopeptides to suppress immune rejection of grafts

Abstract

The present specification discloses the use and method of using a gem-difluorinated C-glycopeptide compound of general formula I, or a pharmaceutically acceptable base, acid addition salt, hydrate or solvate of a compound of general formula I, for suppressing or preventing immune rejection of an isolated graft that has been contacted with said compound prior to transplantation into a subject in need thereof.

Description

[Technical Field]

[0001] Cross-reference of related applications. This application claims priority to U.S. Provisional Patent Application No. 62 / 965,289, filed on 24 January 2020, and U.S. Provisional Patent Application No. 63 / 118,715, filed on 26 November 2020, the specifications of which are herein by reference in their entirety.

[0002] The subject matter disclosed generally relates to methods for transplanting grafts, and more specifically, to methods for suppressing or preventing immune rejection of grafts transplanted into subjects requiring transplantation after discontinuation of immunosuppressant therapy following transplantation. [Background technology]

[0003] Antifreeze biocompounds, and especially glycoproteins, are found in the natural environment. These compounds are present in some fish species, for example, and enable them to survive in low-temperature environments (i.e., near-zero or below-freezing temperatures). Scientists have studied how antifreeze compounds extracted from the natural environment (fish, amphibians, plants, insects, etc.) affect these phenomena. Research has focused on synthesizing analogous compounds that are sufficiently stable for commercial applications and whose activity is at least equivalent to or greater than that of the natural molecules.

[0004] Antifreeze proteins (AFPs) are attracting increasing attention due to their ability to protect cells under various conditions. They are naturally present in fish in the Arctic and Antarctic regions, as well as invertebrates living in other cold climates, and are involved in maintaining cellular and tissue function even below freezing temperatures. AFPs were successfully isolated in the 1950s and have been demonstrated to have the ability to non-coagulate the freezing temperature of body fluids by binding to ice crystals.

[0005] Early experiments using these compounds in the field of organ and tissue transplantation have shown promising results, making them attractive therapeutic candidates for protecting cells from adverse conditions associated with the recovery-preservation-reperfusion process. Furthermore, in the cryopreservation of various cells, including islets of Langerhans, it has been demonstrated that supplementing with AFP during cryopreservation significantly improves their viability and function. The anti-aging glycopeptide (AAGP) used in this invention TM This stems from attempts to obtain analogues of antifreeze glycoproteins.

[0006] Anti-aging glycopeptide (AAGP) TM The compound is a gem-difluorinated C-glycopeptide that has been proposed to be applicable under severe cellular stress conditions such as nutrient deficiency, high temperature / freezing storage, oxidative stress caused by hydrogen peroxide (H2O2), ultraviolet irradiation, and inflammation. [Overview of the Initiative] [Problems that the invention aims to solve]

[0007] Cell, organ (or part thereof), and tissue transplantation are promising approaches to replace tissue lost due to disease. However, the long-term survival and functional integration rates of transplanted cells, organs, or tissues remain problematic, due to immune rejection disorders that cause immune rejection reactions, and the toxicity of immunosuppressants that cause loss of function in the graft.

[0008] Therefore, in this field, there is a need for means to improve graft survival. [Means for solving the problem]

[0009] According to this embodiment, a method for suppressing or preventing immune rejection of a graft, wherein, before transplantation into a subject in need, the isolated graft is subjected to general formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 represents H, AA1, or AA1 - AA2, R 5 represents OH, AA1, or AA1 - AA2, AA1 and AA2 independently represent amino acids having non - polar side chains, and R 1 , R 2 , R 3 are independent groups, where R 1 , R 2 and R 3 two of which are selected from H, CH 3、 CH2Ph, CH(CH3) 2、 CH2CH(CH3)2 or CH(CH3)CH2CH3, and the remaining R 1 , R 2 , R 3 is [Chemical formula] [wherein, n is an integer from 3 to 4, Y, Y’ are independent groups, where Y, Y’ represent H, OR, N3, NR’R” or SR’’’, where R represents H, benzyl group, trimethylsilyl group, tert - butyldimethylsilyl group, tert - butyldiphenylsilyl group or acetate group, R’, R” independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group or C(=O)-Bn, R’’’ represents H, alkyl group, or acetate group, R 6 represents H, CH3, CH2OH, CH2 - glycoside group or CH2 - OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert - butyldimethylsilyl group, tert - butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8represents a hydrogen atom H or a free or protected alcohol functional group.} represents a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I with a base or acid. The process of bringing it into contact: b) A subject requiring the graft is receiving immunosuppressant therapy to inhibit or prevent immune rejection of the graft, and the step is to implant the graft into the subject requiring the graft. c) discontinuing the immunosuppressant therapy after a sufficient amount of time has elapsed for the transplantation of the graft into the subject who requires it. The present invention provides a method for suppressing or preventing immune rejection of a graft, including the present invention.

[0010] The above method involves step a) before step a'): a') Steps to isolate the graft This may further include:

[0011] The immunosuppressant may be sirolimus, tacrolimus, cyclosporine, everolimus, or a combination thereof.

[0012] The test subject may be a human subject.

[0013] Isolated grafts can be isolated from living donors, cadaveric donors, or a combination thereof.

[0014] The compound represented by formula I is represented by formula II: [ka] {In the formula, N is an integer from 1 to 5, and R 1 , R 2 , R 3 It is an independent group, Here, R 1 , R 2 and R 3 Two of them are selected from H, CH3, and the remaining R 1, R 2 and R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 Here, OH, OGP', NH2, N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. ] represents and R 1 and R 2 However, if it is H or CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', or SR''''. Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 OH, OGP'NH 2、 N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H or CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', and SR''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 R is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), 7 is represented by OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H or CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom or a freed or protected alcohol functional group. It may be a compound represented by}.

[0015] The compound represented by formula I is, formula III: [ka] It may be a compound represented by [formula].

[0016] Contact of isolated grafts may be with compounds represented by formula I, formula II, or formula III in concentrations ranging from approximately 0.01 mg / mL to approximately 5 mg / mL.

[0017] Contact with isolated grafts may involve contact with compounds represented by formula I, formula II, or formula III at concentrations ranging from approximately 1 mg / mL to approximately 5 mg / mL.

[0018] Isolated grafts are organs, organ fragments, tissues, and isolated tissues. done Includes cells, or combinations thereof.

[0019] The organ or organ fragment may be the liver, heart, kidney, lung, pancreas, intestine, thymus, uterus, stomach, testes, penis, or hand.

[0020] Tissues include bone, bone marrow, tendons, cornea, heart valves, skin, or It could be a blood vessel.

[0021] Isolation done Cells are isolated. done Pancreatic cells, and isolation done It may be one of the following: pancreatic progenitor cells, adult stem cells, neuronal sensory progenitor cells, reticular cells, glial cells, nerve cells, cardiomyocytes, hepatocytes, or hematopoietic stem cells.

[0022] Isolation done Pancreatic cells are isolated. done α cells, isolated done beta cells, isolated done δ cells, isolated done They may be gamma cells, epsilon cells, or a combination thereof.

[0023] Isolation done Pancreatic cells are isolated. done It could be a beta cell.

[0024] Neuronal sensory progenitor cells may be photoreceptor progenitor cells.

[0025] The graft can be exposed to the compound for approximately 1 minute to 1 hour before transplantation.

[0026] In the method of the present invention, the organ may be the pancreas, and the method may be a method for treating diabetes.

[0027] In the method of the present invention, the isolated cells may be neuronal sensory progenitor cells, and this method may be used for the treatment of retinal degenerative diseases.

[0028] Retinal degenerative diseases may include age-related macular degeneration (AMD), retinitis pigmentosa (RP), retinal vasculitis, or sarcoidosis.

[0029] The isolated grafts may be washed to remove compounds represented by formulas I, II, or III.

[0030] According to other embodiments, General Formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7represents OH, OGP’, NH2, N3, NHGP’ or NGP’GP” (where GP’ and GP” are independently selected from an alkyl group, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 is [Chemical formula] [wherein, n is an integer from 3 to 4, Y and Y’ are independent groups, where Y and Y’ represent H, OR, N3, NR’R” or SR’’’, where R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group or an acetate group, R’ and R” independently represent H, an alkyl group, an allyl group, a benzyl group, a tosylate group, a C(=O)-alkyl group or C(=O)-Bn, R’’’ represents H, an alkyl group, or an acetate group, R 6 is H, CH3, CH2OH, a CH2-glycoside group or CH2-OGP (where GP represents a protecting group selected from an alkyl group, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group). R 7 represents OH, OGP’, NH2, N3, NHGP’ or NGP’GP” (where GP’ and GP” are independently selected from an alkyl group, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group). R 8 represents a hydrogen atom H or a free or protected alcohol functional group.], and R 2 and R 3 when represent H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 is [Chemical formula] [wherein, n is an integer from 3 to 4, Y and Y’ are independent groups, where, Y and Y’ represent H, OR, N3, NR’R” or SR’’’, where, R represents H, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group or acetate group, R’ and R” independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group or C(=O)-Bn, R’’’ represents H, alkyl group, or acetate group, R 6 is H, CH3, CH2OH, CH2-glycoside group or CH2-OGP (where, GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group).), R 7 is OH, OGP’, NH2, N3, NHGP’ or NGP’GP” (where, GP’ and GP” are independently selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group).), R 8represents a hydrogen atom H or a free or protected alcohol functional group.} The use of a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, may provide a use to suppress or prevent immunorejection of isolated grafts that have come into contact with these compounds before implantation into a subject requiring such use.

[0031] Subjects requiring this treatment may be receiving immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0032] Immunosuppressant therapy may be discontinued after sufficient time has elapsed to transplant the graft into the subject who requires it.

[0033] The immunosuppressant may be sirolimus, tacrolimus, cyclosporine, everolimus, or a combination thereof.

[0034] The test subject may be a human subject.

[0035] Isolated grafts can be isolated from living donors, cadaveric donors, or a combination thereof.

[0036] The compound represented by formula I is represented by formula II: [ka] {In the formula, N is an integer from 1 to 5, and R 1 , R 2 , R 3 It is an independent group, Here, R 1 , R 2 and R 3 Two of them are selected from H, CH3, and the remaining R 1 , R 2 and R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 Here, OH, OGP', NH2, N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. ] represents and R 1 and R 2 However, if it is H or CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', or SR''''. Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 OH, OGP'NH 2、 N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H or CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', and SR''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 R is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), 7 is represented by OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H or CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R7 represents OH, OGP’, NH2, N3, NHGP’ or NGP’GP” (where GP’ and GP” are independently selected from an alkyl group, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group). ), and R 8 represents a hydrogen atom or a free or protected alcohol functional group. It can be a compound represented by {}.

[0037] The compound represented by formula I can be a compound represented by formula III:

Chemical formula

[0038] The contact of the isolated graft can be made with a compound represented by formula I, formula II or formula III at about 0.01 mg / mL to about 5 mg / mL.

[0039] The contact of the isolated graft can be made with a compound represented by formula I, formula II or formula III at about 1 mg / mL to about 5 mg / mL.

[0040] The isolated graft includes an organ, a fragment of an organ, tissue, isolated cells, or a combination thereof.

[0041] The organ or the fragment of the organ can be a liver, a heart, a kidney, a lung, a pancreas, an intestine, a thymus, a uterus, a stomach, a testis, a penis, a hand.

[0042] The tissue can be bone, bone marrow, tendon, cornea, heart valve, skin, and blood vessel.

[0043] The isolated cells can be any one of isolated pancreatic cells, isolated pancreatic progenitor cells, adult stem cells, neural sensory progenitor cells, reticular cells, glial cells, nerve cells, cardiomyocytes, hepatocytes, and hematopoietic stem cells.

[0044] Isolated pancreatic cells may be isolated α cells, isolated β cells, isolated δ cells, isolated γ cells, ε cells, or a combination thereof.

[0045] Isolated pancreatic cells may be isolated β-cells.

[0046] Neuronal sensory progenitor cells may be photoreceptor progenitor cells.

[0047] The isolated grafts can be exposed to the compound for approximately 1 minute to 1 hour before transplantation.

[0048] The isolated grafts can be washed to remove compounds represented by formulas I, II, or III.

[0049] In the use of this invention, the organ may be the pancreas, and the use may be for the treatment of diabetes.

[0050] In the use of the present invention, the isolated cells may be neurosensory progenitor cells, and the use may be for the treatment of retinal degenerative diseases.

[0051] Retinal degenerative diseases may include age-related macular degeneration (AMD), retinitis pigmentosa (RP), retinal vasculitis, or sarcoidosis.

[0052] According to other embodiments, a) Before transplanting it to a subject who needs it, the isolated graft should be prepared using General Formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3is an independent group, where R 1 , R 2 and R 3 two of which are selected from H, CH 3、 CH2Ph, CH(CH3) 2、 CH2CH(CH3)2 or CH(CH3)CH2CH3, and the remaining R 1 , R 2 , R 3 is [Chemical formula] [wherein, n is an integer from 3 to 4, Y and Y' are independent groups, where Y and Y' represent H, OR, N3, NR'R" or SR''', where R represents H, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group or acetate group, R' and R" independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group or C(=O)-Bn, R''' represents H, alkyl group, or acetate group, R 6 represents H, CH3, CH2OH, CH2-glycoside group or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group).), R 7 represents OH, OGP', NH2, N3, NHGP' or NGP'GP" (where GP' and GP" are independently selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group).), R 8 represents a hydrogen atom H, or a free or protected alcohol functional group.], and R 1 and R 2However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group.} represents a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, b) The step of implanting the graft into the subject who has not received, or requires, immunosuppressive drug treatment to inhibit or prevent immune rejection of the graft, The present invention provides a method for inhibiting or preventing immune rejection, including the following:

[0053] According to other embodiments, General formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 The two are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8The present invention provides a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, for use in inhibiting or preventing immunorejection of isolated grafts that have been in contact with these compounds before implantation into a subject requiring the compound.

[0054] Subjects requiring this treatment may be receiving immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0055] Immunosuppressant therapy may be discontinued after sufficient time has elapsed to transplant the graft into the subject who requires it.

[0056] According to other embodiments, before implanting it into a subject requiring it, General Formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 The present invention provides a graft for suppressing or preventing immunorejection of an isolated graft that has been contacted with a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, wherein} represents a hydrogen atom H or a free or protected alcohol functional group.

[0057] Subjects requiring this treatment may be receiving immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0058] Immunosuppressant therapy may be discontinued after sufficient time has elapsed to transplant the graft into the subject who requires it.

[0059] The following terms are defined as follows:

[0060] As used herein, the term “immunosuppression” is intended to mean activation or reduction of the effectiveness of the immune system. Parts of the immune system itself may have an immunosuppressive effect on other parts of the immune system, and immunosuppression may occur as a side effect of treating other diseases. However, in this invention… In a more specific context, immunosuppression is performed to prevent the body from rejecting organ transplants through immune rejection disorders. Individuals who are immunosuppressed, or whose immune systems are weakened for other reasons, are considered immunocompromised.

[0061] As used herein, the term “discontinuation” in relation to the discontinuation of immunosuppressant therapy is intended to mean ceasing the provision of the immunosuppressant. In the context of the present invention, this is after a sufficient amount of time has elapsed for the transplantation of the graft as defined below.

[0062] As used herein, the term “graft” is intended to mean a living organ, part thereof, tissue, and / or cells that are surgically transplanted. Types of grafts generally include autografts (e.g., skin grafts), which are grafts taken from a part of one individual's body and transplanted to another part of the same individual; homogeneous grafts (e.g., grafts between identical twins), which are grafts taken from one individual and transplanted to another individual of the same genetic makeup; allografts, which are grafts taken from one individual and transplanted to another individual of the same species that is not genetically identical; and xenografts, which are grafts taken from one individual and transplanted to an individual of a different species (e.g., from an animal to a human). In the context of the present invention, grafts usually refer to allografts and xenografts that are subject to immune rejection. According to one embodiment, grafts may be cryopreserved, for example, by freezing in liquid nitrogen. According to other different embodiments, grafts may be cryopreserved in any form and / or not frozen at all. Therefore, the method or use of the present invention will not involve the step of cryopreserving isolated grafts.

[0063] As used herein, the terms “graft transplant” and “transplant” are intended to mean the insertion, fixation, or attachment of a graft to the human body.

[0064] As used herein, the term “sufficient time for transplantation” is intended to mean the time it takes for the transplanted graft to adhere to and / or fix in the human body. This time varies and differs depending on the type of graft being transplanted. It can range from minutes, hours, a day or days, a week or weeks or even months. Sufficient time is approximately 1 minute to 10 minutes, or approximately 1 minute to 15 minutes, or approximately 1 minute to 30 minutes, or approximately 1 minute to 45 minutes, or approximately 1 minute to 1 hour, or approximately 1 minute to 2 hours, or approximately 1 minute to 6 hours, or approximately 1 minute to 12 hours, or approximately 1 minute to 18 hours, or approximately 1 minute to 1 day, or approximately 1 minute to 2 days, or approximately 1 minute to 3 days, or approximately 1 minute to 4 days, or approximately 1 minute to 5 days, or approximately 1 minute to 6 days, or approximately 1 minute to 1 week, or approximately 1 minute to 2 weeks, or approximately 1 minute to 3 Weeks, or about 1 minute to about 1 month, or about 1 minute to about 2 months, or about 1 minute to about 3 months, or about 1 minute to about 4 months, or about 1 minute to about 5 months, or about 1 minute to about 6 months, or about 10 minutes to about 15 minutes, or about 10 minutes to about 30 minutes, or about 10 minutes to about 45 minutes, or about 10 minutes to about 1 hour, or about 10 minutes to about 2 hours, or about 10 minutes to about 6 hours, or about 10 minutes to about 12 hours, or about 10 minutes to about 18 hours, or about 10 minutes to about 1 day, or about 10 minutes to about 2 days, or about 10 minutes to about 3 days, or about 10 minutes to about 4 days, or about 10 minutes to about 5 days, or about 10 minutes to about 6 days, or about 10 minutes to about 1 week, or about 10 minutes to about 2 weeks, or about 10 minutes to about 3 weeks, or about 10 minutes to about 1 month, or about 10 minutes to about 2 months, or about 10 minutes to about 3 months, or about 10 minutes to about 4 months, or about 10 minutes to about 5 months, or about 10 minutes to about 6 months, or about 15 minutes to about 30 minutes, or about 15 minutes to about 45 minutes, or about 15 minutes to about 1 hour, or about 15 minutes to about 2 hours, or about 15 minutes to about 6 hours, or about 15 minutes to about 12 hours, or about 15 minutes to about 18 hours, or about 15 minutes to about 1 day, or about 15 minutes to about 2 days, or about 15 minutes to about 3 days, or about 15 minutes to about 4 days, or about 15 minutes to about 5 days, or about 15 minutes to about 6 days, or about 15 minutes to about 1 week, or about 15 minutes to about 2 weeks, or about 15 minutes to about 3 weeks, or about 15 minutes to about 1 month, or about 15 minutes to about 2 months, or about 15 minutes to about 3 months, or about 15 minutes to about 4 months, or about 15 minutes to about 5 months, or about 15 minutes to about 6Months, or approximately 30 minutes to approximately 45 minutes, or approximately 30 minutes to approximately 1 hour, or approximately 30 minutes to approximately 2 hours, or approximately 30 minutes to approximately 6 hours, or approximately 30 minutes to approximately 12 hours, or approximately 30 minutes to approximately 18 hours, or approximately 30 minutes to approximately 1 day, or approximately 30 minutes to approximately 2 days, or approximately 30 minutes to approximately 3 days, or approximately 30 minutes to approximately 4 days, or approximately 30 minutes to approximately 5 days, or approximately 30 minutes to approximately 6 days, or approximately 30 minutes to approximately 1 week, or approximately 30 minutes to approximately 2 weeks, or approximately 30 minutes to approximately 3 weeks, or approximately 30 minutes to approximately 1 month, or approximately 30 minutes to approximately 2 months, or approximately 30 minutes to approximately 3 months, or approximately 30 minutes to approximately 4 months, or approximately 3 0 minutes to approximately 5 months, or approximately 30 minutes to approximately 6 months, or approximately 45 minutes to approximately 1 hour, or approximately 45 minutes to approximately 2 hours, or approximately 45 minutes to approximately 6 hours, or approximately 45 minutes to approximately 12 hours, or approximately 45 minutes to approximately 18 hours, or approximately 45 minutes to approximately 1 day, or approximately 45 minutes to approximately 2 days, or approximately 45 minutes to approximately 3 days, or approximately 45 minutes to approximately 4 days, or approximately 45 minutes to approximately 5 days, or approximately 45 minutes to approximately 6 days, or approximately 45 minutes to approximately 1 week, or approximately 45 minutes to approximately 2 weeks, or approximately 45 minutes to approximately 3 weeks, or approximately 45 minutes to approximately 1 month, or approximately 45 minutes to approximately 2 months, or approximately 45 minutes to approximately 3 months, or approximately 45 minutes to approximately 4 months Month, or approximately 45 minutes to approximately 5 months, or approximately 45 minutes to approximately 6 months, or approximately 1 hour to approximately 2 hours, or approximately 1 hour to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 18 hours, or approximately 1 hour to approximately 1 day, or approximately 1 hour to approximately 2 days, or approximately 1 hour to approximately 3 days, or approximately 1 hour to approximately 4 days, or approximately 1 hour to approximately 5 days, or approximately 1 hour to approximately 6 days, or approximately 1 hour to approximately 1 week, or approximately 1 hour to approximately 2 weeks, or approximately 1 hour to approximately 3 weeks, or approximately 1 hour to approximately 1 month, or approximately 1 hour to approximately 2 months, or approximately 1 hour to approximately 3 months, or approximately 1 hour to approximately 4 months, or approximately 1 hour Intervals to approximately 5 months, or approximately 1 hour to approximately 6 months, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 18 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 1 month, or approximately 2 hours to approximately 2 months, or approximately 2 hours to approximately 3 months, or approximately 2 hours to approximately 4 months, or approximately 2 hours to approximately 5 months, or approximately 2 hours to approximately 6 months,Or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 18 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 1 month, or approximately 6 hours to approximately 2 months, or approximately 6 hours to approximately 3 months, or approximately 6 hours to approximately 4 months, or approximately 6 hours to approximately 5 months, or approximately 6 hours to approximately 6 months, or approximately 12 hours to approximately 18 hours, or approximately 12 hours to approximately 1 day, or approximately 12 hours Intervals to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 1 month, or approximately 12 hours to approximately 2 months, or approximately 12 hours to approximately 3 months, or approximately 12 hours to approximately 4 months, or approximately 12 hours to approximately 5 months, or approximately 12 hours to approximately 6 months, or approximately 18 hours to approximately 1 day, or approximately 18 hours to approximately 2 days, or approximately 18 hours to approximately 3 days, or approximately 18 hours to approximately 4 days, or approximately 18 hours to approximately 5 days, or Approximately 18 hours to approximately 6 days, or approximately 18 hours to approximately 1 week, or approximately 18 hours to approximately 2 weeks, or approximately 18 hours to approximately 3 weeks, or approximately 18 hours to approximately 1 month, or approximately 18 hours to approximately 2 months, or approximately 18 hours to approximately 3 months, or approximately 18 hours to approximately 4 months, or approximately 18 hours to approximately 5 months, or approximately 18 hours to approximately 6 months, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 1 month, or approximately 1 day to approximately 2 months, or approximately 1 day to approximately 3 months, or approximately 1 day to approximately 4 months, or approximately 1 day to approximately 5 months, or approximately 1 day to approximately 6 months, or approximately 2 days to approximately 3 days, or approximately 2 days to approximately 4 days, or approximately 2 days to approximately 5 days, or approximately 2 days to approximately 6 days, or approximately 2 days to approximately 1 week, or approximately 2 days to approximately 2 weeks, or approximately 2 days to approximately 3 weeks, or approximately 2 days to approximately 1 month, or approximately 2 days to approximately 2 months, or approximately 2 days to approximately 3 months, or approximately 2 days to approximately 4 months, or approximately 2 days to approximately 5 months, or approximately 2 days to approximately 6 months, or approximately 3 days to approximately 4 days, or approximately 3 days to approximately 5 days, or approximately 3 days to approximately 6 days, or approximately 3 days to approximately 1 week, or approximately 3 days to approximately 2 weeks,Or approximately 3 days to approximately 3 weeks, or approximately 3 days, up to about 1 month, or about 3 days to about 2 months, or about 3 days to about 3 months, or about 3 days to about 4 months, or about 3 days to about 5 months, or about 3 days to about 6 months, or about 4 days to about 5 days, or about 4 days to about 6 days, or about 4 days to about 1 week, or about 4 days to about 2 weeks, or about 4 days to about 3 weeks, or about 4 days to about 1 month, or about 4 days to about 2 months, or about 4 days to about 3 months, or about 4 days to about 4 months, or about 4 days to about 5 months, or about 4 days to about 6 months, or about 5 days to about 6 days, or about 5 days to about 1 week, Or approximately 5 days to approximately 2 weeks, or approximately 5 days to approximately 3 weeks, or approximately 5 days to approximately 1 month, or approximately 5 days to approximately 2 months, or approximately 5 days to approximately 3 months, or approximately 5 days to approximately 4 months, or approximately 5 days to approximately 5 months, or approximately 5 days to approximately 6 months, or approximately 6 days to approximately 1 week, or approximately 6 days to approximately 2 weeks, or approximately 6 days to approximately 3 weeks, or approximately 6 days to approximately 1 month, or approximately 6 days to approximately 2 months, or approximately 6 days to approximately 3 months, or approximately 6 days to approximately 4 months, or approximately 6 days to approximately 5 months, or approximately 6 days to approximately 6 months, or approximately 1 week to approximately 2 weeks, or approximately 1 week. Up to approximately 3 weeks, or approximately 1 week to approximately 1 month, or approximately 1 week to approximately 2 months, or approximately 1 week to approximately 3 months, or approximately 1 week to approximately 4 months, or approximately 1 week to approximately 5 months, or approximately 1 week to approximately 6 months, or approximately 2 weeks to approximately 3 weeks, or approximately 2 weeks to approximately 1 month, or approximately 2 weeks to approximately 2 months, or approximately 2 weeks to approximately 3 months, or approximately 2 weeks to approximately 4 months, or approximately 2 weeks to approximately 5 months, or approximately 2 weeks to approximately 6 months, or approximately 3 weeks to approximately 1 month, or approximately 3 weeks to approximately 2 months, or approximately 3 weeks to approximately 3 months, or approximately 3 weeks to approximately 4 months, or approximately 3 weeks to approximately It may be 5 months, or about 3 weeks to about 6 months, or about 1 month to about 2 months, or about 1 month to about 3 months, or about 1 month to about 4 months, or about 1 month to about 5 months, or about 1 month to about 6 months, or about 2 months to about 3 months, or about 2 months to about 4 months, or about 2 months to about 5 months, or about 2 months to about 6 months, or about 3 months to about 4 months, or about 3 months to about 5 months, or about 3 months to about 6 months, or about 4 months to about 5 months, or about 4 months to about 6 months, or about 5 months to about 6 months.

[0065] As used herein, the terms “host-versus-graft disease” and “graft-versus-host disease” refer to syndromes characterized by inflammation in different organs that are commonly associated with all types of transplants of a transplanted subject, including solid organ transplants, some solid organ transplants, cell transplants such as bone marrow transplants and stem cell transplants, and pancreatic cell transplants, in which either the graft or the host triggers an immune response against the host or the graft, respectively.

[0066] As used herein, the terms “transplant” or “organ transplant” refer to a medical procedure in which an organ is removed from one body and placed in the body of a recipient in order to replace a damaged or missing organ, part thereof, tissue, and / or cells. The donor and recipient may be in the same location, or the organ may be transported from the donor's location to another location. An organ, part thereof, tissue, and / or cells transplanted within the body of the same person is called an autologous transplant. A recent transplant between two subjects of the same species is called an allogeneic transplant. Allogeneic grafts may come from a living or cadaveric source.

[0067] As used herein in the context of the present invention, the terms “inhibit,” “inhibition,” or “inhibiting” mean to delay, hinder, suppress, reduce, delay, protect, or prevent. For example, “inhibition of host-versus-graft disease,” “inhibition of graft-versus-host disease,” or “inhibition of immune rejection” of an organ, part thereof, tissue, and / or cell means, as used herein, to delay, hinder, suppress, reduce, reduce, protect, or prevent the onset of host-versus-graft / graft-versus-host disease and / or immune rejection and / or immune rejection.

[0068] As used herein, the terms “immune rejection” or “transplant rejection” refer to the case in which a transplanted organ, part thereof, tissue, and / or cells are rejected by the recipient’s immune system and destroyed. Transplant rejection can be mitigated by determining the molecular similarity between the donor and recipient and by using immunosuppressants after transplantation.

[0069] As used herein, the term “contact” means bringing an organ, a part thereof, a tissue, and / or cells into contact with the compound of the present invention for a sufficient time to provide the effect conferred by the compound. In some embodiments, contact is intended to mean incubating an organ, a part thereof, a tissue, and / or cells with a suitable solution containing the compound. In some embodiments, this may include perfusion of the organ, a part thereof, or tissue (i.e., passage of blood, a blood substitute, or other fluid through blood vessels or other natural channels within the organ or tissue).

[0070] As used herein, the term “organ” is intended to mean a part of an organism that is generally self-contained and possesses a specific life function, such as the human heart or liver.

[0071] The term “subject” preferably refers to a human subject, but may also refer to any mammal, including animal models. Mammals of interest include, but are not limited to, rodents such as mice and rats, livestock such as pigs, horses, and cattle, pets such as dogs and cats, and primates. Subjects may also be referred to as “patients” in this specification.

[0072] As used herein, the term “composition” is intended to encompass products containing specific amounts of specific components, and any products resulting directly or indirectly from specific amounts of specific components in combination. In relation to pharmaceutical compositions or other general compositions, the term is intended to encompass products containing active and inactive components constituting a carrier, and any products resulting directly or indirectly from any combination, complexation, or aggregation of any two or more components, from the dissociation of one or more components, or from other types of reactions or interactions of one or more components. Accordingly, general pharmaceutical compositions or other compositions of the present invention encompass any compositions prepared by mixing the compounds of the present invention with pharmaceutically acceptable carriers. “Pharmaceutically acceptable” or “acceptable” means that the carrier, diluent, or excipient is compatible with the other components of the formulation and is not harmful to its recipient.

[0073] The term “neurosensory progenitor cells” is intended to mean cells derived from human embryonic stem cells or induced pluripotent stem cells, isolated by known protocols and / or as described below herein, and cells grown in vivo, ex vivo, and / or in vitro.

[0074] The terms “pancreatic cells,” “isolated pancreatic cells,” “islet cells,” or “isolated islet cells” are intended to mean cells derived from the islets of Langerhans of the pancreas from living or deceased donors isolated through known protocols and / or as described below herein, as well as cells derived from the pancreas grown in vivo, ex vivo, and / or in vitro. Cells of the islets of Langerhans include α-cells, which produce the hormone glucagon and account for approximately 15-20% of islet cells; β-cells, which produce the hormones insulin and amylin and account for approximately 65-80% of islet cells; δ-cells, which produce the hormone somatostatin and account for approximately 3-10% of islet cells; PP-cells (also known as γ-cells), which produce the hormone pancreatic polypeptide (3-5% of islet cells); and ε-cells, which produce the hormone ghrelin and account for 1% of islet cells.

[0075] The terms “pancreatic β-cells” and “isolated pancreatic β-cells” are intended to mean cells derived from the pancreas of a living or deceased donor, isolated through known protocols and / or as described below herein, and pancreatic β-cells grown in vivo, ex vivo, and / or in vitro.

[0076] The terms “pancreatic progenitor cells” and “isolated pancreatic progenitor cells” are intended to mean cells derived from any suitable source, such as embryonic stem cells (of human or other origin), that are differentiated or have been differentiated into pancreatic progenitor cells in vivo, ex vivo, or in vitro using a known protocol. Isolated pancreatic progenitor cells may become pancreatic β-cells by further spontaneous differentiation or by treatment using a known protocol, in vivo, ex vivo, or in vitro. For example, isolated pancreatic progenitor cells may be obtained in vitro using a differentiation protocol and then further differentiated into pancreatic β-cells in vitro using a differentiation protocol. Alternatively, isolated pancreatic progenitor cells may be obtained in vitro using a differentiation protocol and then further differentiated into pancreatic β-cells in vivo after implantation / transplantation to a patient in need.

[0077] "Alkyl group," and other groups having the prefix "alk," such as alkoxy and alkanoyl groups, means a carbon chain that may be linear, branched, or a combination thereof, unless otherwise defined. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, and nonyl groups. If the specified number of carbon atoms allows, for example, C3-10, the term alkyl includes cycloalkyl groups and combinations of linear or branched alkyl chains with cycloalkyl structures. If the number of carbon atoms is not specified, C1-6 is intended.

[0078] A "cycloalkyl group" is a subset of an alkyl group, meaning a saturated carbon ring having a specified number of carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. Cycloalkyl groups are generally monocyclic unless otherwise specified. Cycloalkyl groups are saturated unless otherwise defined.

[0079] The term "alkoxy group" refers to a linear or branched alkoxide with a specified number of carbon atoms (e.g., a C1-6 alkoxy group) or any number within this range [i.e., a methoxy group (MeO-), an ethoxy group, an isopropoxy group, etc.].

[0080] The term "alkylthio group" refers to a linear or branched alkyl sulfide [i.e., methylthio group (MeS-), ethylthio group, isopropylthio group, etc.] with a specified number of carbon atoms (e.g., C1-6 alkylthio group) or any number within this range.

[0081] The term "alkylamino group" refers to a linear or branched alkylamine with a specified number of carbon atoms (e.g., C1-6 alkylamino) or any number within this range [i.e., methylamino group, ethylamino group, isopropylamino group, t-butylamino group, etc.].

[0082] The term "alkylsulfonyl group" refers to a linear or branched alkylsulfone with a specified number of carbon atoms (e.g., C1-6 alkylsulfonyl group), or any number within this range [i.e., methylsulfonyl group (MeSO2-), ethylsulfonyl group, isopropylsulfonyl group, etc.].

[0083] The term "alkylsulfinyl group" refers to a linear or branched alkyl sulfoxide with a specified number of carbon atoms (e.g., C1-6 alkylsulfinyl group), or any number within this range [i.e., methylsulfinyl group (MeSO-), ethylsulfinyl group, isopropylsulfinyl group, etc.].

[0084] "Alkyloxycarbonyl group" means a linear or branched ester of the carboxylic acid derivative of the present invention having a specified number of carbon atoms (e.g., a C1-6 alkyloxycarbonyl group), or any number within this range [i.e., a methyloxycarbonyl group (MeOCO-), an ethyloxycarbonyl group, or a butyloxycarbonyl group].

[0085] An "aryl group" refers to a monocyclic or polycyclic aromatic ring system containing a carbocyclic atom. Preferred aryl groups are monocyclic or bicyclic 6-10 membered aromatic ring systems. Phenyl and naphthyl groups are preferred aryl groups. The most preferred aryl group is the phenyl group.

[0086] A "heterocycle" refers to a saturated or unsaturated non-aromatic ring or cyclic system containing at least one heteroatom selected from O, S, and N, and further containing sulfur oxidants, i.e., SO and SO2. Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1,4-dioxane, morpholine, 1,4-dithiane, piperazine, piperidine, 1,3-dioxolane, imidazolidine, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane, 1,3-dioxane, 1,3-dithiane, oxathiane, thiomorpholine, 2-oxopiperidine-1-yl, 2-oxopyrrolidine-1-yl, 2-oxazetidine-1-yl, 1,2,4-oxadiazine-5(6H)-on-3-yl, etc.

[0087] "Heteroaryl" refers to an aromatic or partially aromatic heterocycle containing at least one ring heteroatom selected from O, S, and N. Therefore, heteroaryls also include heteroaryls fused to other types of rings, such as aryl, cycloalkyl, and non-aromatic heterocycles. Examples of heteroaryl groups include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl (especially 1,3,4-oxadiazole-2-yl and 1,2,4-oxadiazole-3-yl), thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furyl, triazinyl, thienyl, pyrimidyl, benzisoxazolyl, benzoxazolyl, benzothiazolyl, and benzothiazolyl. These include zolyl groups, dihydrobenzofuranyl groups, indolinyl groups, pyridadinyl groups, indazolyl groups, isoindolyl groups, dihydrobenzothienyl groups, indolidinyl groups, cinolinyl groups, phthalazinyl groups, quinazolinyl groups, naphthilidinyl groups, carbazolyl groups, benzodioxolyl groups, quinoxalinyl groups, prinyl groups, fluzanyl groups, isobenzylfuranyl groups, benzimidazolyl groups, benzofuranyl groups, benzothienyl groups, quinolyl groups, indolyl groups, isoquinolyl groups, and dibenzofuranyl groups. Heterocyclyl groups and heteroaryl groups include rings and ring systems containing 3 to 15 atoms, forming 1 to 3 rings.

[0088] "Halogen" refers to fluorine, chlorine, bromine, and iodine. Chlorine and fluorine are generally preferred. When the halogen is substituted on an alkyl or alkoxy group, fluorine is most preferred (e.g., CF3O and CF3CH2O).

[0089] Before describing the present invention in detail, several terms are defined. As used herein, the singular forms "a," "an," and "the" include multiple references unless the context clearly indicates otherwise.

[0090] It should be noted that terms such as “preferably,” “generally,” and “typically” are not used herein to limit the scope of the claimed invention or to imply that certain features are significant, essential, or even more important to the structure or function of the claimed invention. Rather, these terms are intended simply to highlight alternative or additional features that may or may not be available in particular embodiments of the invention.

[0091] For the purposes of describing and defining the present invention, the term “substantially” is used herein to describe the degree of inherent uncertainty that may be attributable to any quantitative comparison, value, measurement, or other expression. Furthermore, herein, “substantially” is used to describe the extent to which a quantitative expression may deviate from the stated standard without altering the fundamental function of the subject matter in question.

[0092] The features and advantages of the subject matter herein will become more apparent in light of the following detailed description of selected embodiments, as shown in the accompanying figures. As should be understood, all disclosed and claimed subject matter is modifiable in various ways without departing from the claims. Therefore, the drawings and description are considered illustrative and not restrictive, and the entire scope of the subject matter is described in the claims.

[0093] Further features and advantages of this disclosure will become apparent from the following detailed description in conjunction with the attached drawings. [Brief explanation of the drawing]

[0094] [Figure 1] Figure 1 is a flowchart of the research design presented in Example 1. [Figure 2] Figure 2 is a flowchart of the procedure used to generate photoreceptor progenitor cells (PPCs) from pluripotent human ES cells (hESCs) used in Example 1. [Figure 3]Figure 3 is a flowchart of the rabbit processing and tissue processing in Example 1. [Figure 4] Figure 4 shows flat-mount images from the control (AAGPTM-free PPC cells) and treatment (AAGPTM-enabled PPC cells) groups 6 months after transplantation. [Figure 5] Figure 5A shows CD4+ T cells in the conjunctival epithelium after induction of acute dry eye disease. Figure 5B shows CD4+ T cells in the conjunctival epithelium after induction of acute dry eye disease.

[0095] Please note that throughout the attached drawings, similar features are identified by the same reference number. [Modes for carrying out the invention]

[0096] Detailed explanation In the embodiment, a) Before transplanting it into a subject who needs it, the isolated graft is prepared using general formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group.} represents a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, b) The subject requiring it is receiving immunosuppressant therapy to suppress or prevent immune rejection of the graft, and the process involves transplanting the graft to the subject requiring it. c) The process of discontinuing immunosuppressant therapy after sufficient time has elapsed for the transplantation of the graft to the subject who requires it. Methods for suppressing or preventing immune rejection, including the following, are disclosed.

[0097] This method further involves step a') before step a). a') Steps to isolate the graft, It may include.

[0098] The immunosuppressant may be sirolimus, tacrolimus, cyclosporine, everolimus, or a combination thereof.

[0099] The test subject may be a human subject.

[0100] The subject shall, in order to suppress or prevent immune rejection of the graft, undergo treatment for at least approximately 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks prior to graft implantation, or approximately 1 to 2 hours, or approximately 1 to 6 hours, or approximately 1 to 12 hours, or approximately 1 hour to 1 day, or approximately 1 hour to 2 days, or approximately 1 hour to 3 days, or approximately 1 hour to 4 days, or approximately 1 hour to 5 days, or approximately 1 hour to 6 days, or approximately 1 hour to 1 week, or approximately 1 hour to 2 weeks, or about 1 hour to about 3 weeks, or about 1 hour to about 4 weeks, or about 2 hours to about 6 hours, or about 2 hours to about 12 hours, or about 2 hours to about 1 day, or about 2 hours to about 2 days, or about 2 hours to about 3 days, or about 2 hours to about 4 days, or about 2 hours to about 5 days, or about 2 hours to about 6 days, or about 2 hours to about 1 week, or about 2 hours to about 2 weeks, or about 2 hours to about 3 weeks, or about 2 hours to about 4 weeks, or about 3 hours to about 12 hours, or about 3 hours to about 1 day, or about 3 hours to about 2 days, or about 3 hours to about 3 days, or about 3 hours to Approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours or approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days,Or approximately 2 to 4 days, or approximately 2 to 5 days, or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 to 4 days, or approximately 3 to 5 days, or approximately 3 to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks, or approximately 3 days to 3 weeks, or approximately 3 days to 4 weeks, or approximately 4 to 5 days, or approximately 4 days to 6 days, or approximately 4 days to 1 week, or approximately 4 days to 2 weeks, or Immunosuppressant treatment may be administered for approximately 4 days to approximately 3 weeks, or approximately 4 days to approximately 4 weeks, or approximately 5 days to approximately 6 days, or approximately 5 days to approximately 1 week, or approximately 5 days to approximately 2 weeks, or approximately 5 days to approximately 3 weeks, or approximately 5 days to approximately 4 weeks, or approximately 6 days to approximately 1 week, or approximately 6 days to approximately 2 weeks, or approximately 6 days to approximately 3 weeks, or approximately 6 days to approximately 4 weeks, or approximately 1 week to approximately 2 weeks, or approximately 1 week to approximately 3 weeks, or approximately 1 week to approximately 4 weeks, or approximately 2 weeks to approximately 3 weeks, or approximately 2 weeks to approximately 4 weeks, or approximately 3 weeks to approximately 4 weeks.

[0101] Isolated grafts can be isolated from living donors, cadaveric donors, or a combination thereof.

[0102] The compound represented by formula I is represented by formula II: [ka] (II) {In the formula, N is an integer from 1 to 5, and R 1 , R 2 , R 3 It is an independent group, Here, R 1 , R 2 and R 3 Two of them are selected from H, CH3, and the remaining R 1 , R 2 and R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 Here, OH, OGP', NH2, N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. ] represents and R 1 and R 2 However, if it is H or CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', or SR''''. Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 It is selected from H, CH3, CH2OH, CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 7 OH, OGP'NH 2、 N3, NHGP', NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H or CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'', and SR''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 R is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), 7 is represented by OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups), R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H or CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 is selected from H, CH3, CH2OH, or CH2-OGP (where GP represents a protecting group selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom or a freed or protected alcohol functional group. It may be a compound represented by}.

[0103] The compound represented by formula I is, formula III: [ka] It may be a compound represented by [formula].

[0104] Contacting the isolated graft may be done using a compound represented by formula I, formula II, or formula III at a concentration of approximately 0.01 mg / mL to approximately 5 mg / mL. Contacting the isolated graft may also be done using a compound represented by formula I, formula II, or formula III at a concentration of approximately 1 mg / mL to approximately 5 mg / mL.

[0105] An isolated graft may include an organ, organ fragment, tissue, isolated cells, or a combination thereof. An organ or organ fragment may be the liver, heart, kidney, lung, pancreas, intestine, thymus, uterus, stomach, testis, penis, or hand. Tissue may be bone, bone marrow, tendon, cornea, heart valve, skin, or blood vessel. Isolated cells may be any one of isolated pancreatic cells, isolated pancreatic progenitor cells, adult stem cells, neurosensory progenitor cells, reticular cells, glial cells, nervous system cells, cardiomyocytes, hepatocytes, or hematopoietic stem cells. Isolated pancreatic cells may be isolated α cells, isolated β cells, isolated δ cells, isolated γ cells, ε cells, or a combination thereof. Isolated pancreatic cells may be isolated β cells. Neurosensory progenitor cells may be photoreceptor progenitor cells. According to one embodiment, the graft may be cryopreserved, for example, by freezing in liquid nitrogen. According to another different embodiment, the graft may not have undergone any form of cryopreservation and / or freezing. Therefore, the method or use of the present invention will not involve the step of cryopreserving isolated grafts.

[0106] The graft should be prepared for at least approximately 1 minute, or approximately 15 minutes, or approximately 30 minutes, or approximately 1 hour, or approximately 6 hours, or approximately 12 hours, or approximately 24 hours, or approximately 48 hours, or approximately 72 hours, or approximately 1 minute to approximately 15 minutes, or approximately 1 minute to approximately 30 minutes, or approximately 1 minute to approximately 1 hour, or approximately 1 minute to approximately 6 hours, or approximately 1 minute to approximately 12 hours, or approximately 1 minute to approximately 24 hours, or approximately 1 minutes to approximately 48 hours, or approximately 1 minute to approximately 72 hours, or approximately 15 minutes to approximately 30 minutes, or approximately 15 minutes to approximately 1 hour, or approximately 15 minutes to approximately 6 hours, or approximately 15 minutes to approximately 12 hours, or approximately 15 minutes to approximately 24 hours, or approximately 15 minutes to approximately 48 hours, or approximately 15 minutes to approximately 72 hours, or approximately 30 minutes to approximately 1 hour, or approximately 30 minutes to approximately 6 hours, or approximately 30 minutes to approximately 12 hours, or approximately 30 minutes to approximately 24 hours, or approximately 30 minutes to approximately 48 hours, or approximately 30 minutes to approximately 72 hours, or approximately 1 hour to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 24 hours, or approximately 1 hour to approximately 48 hours, or approximately 1 hour to approximately 72 hours The compound may be exposed to the following conditions: for a period of time, or approximately 2 to 12 hours, or approximately 2 to 24 hours, or approximately 2 to 48 hours, or approximately 2 to 72 hours, or approximately 6 to 12 hours, or approximately 6 to 24 hours, or approximately 6 to 48 hours, or approximately 6 to 72 hours, or approximately 12 to 24 hours, or approximately 12 to 48 hours, or approximately 12 to 72 hours, or approximately 24 to 48 hours, or approximately 24 to 72 hours, or approximately 1 minute, 15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, or 72 hours.

[0107] The period after discontinuation of immunosuppressant therapy, or in other words, sufficient time for transplantation, is at least 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks or 4 weeks after graft transplantation, or about 1 hour to about 2 hours, or about 1 hour to about 6 hours, or about 1 hour to about 12 hours, or about 1 hour to about 1 day, or about 1 hour to about 2 days, or about 1 hour to about 3 days, or about 1 hour to about 4 days, or about 1 hour to about 5 days, or about 1 hour to about 6 days, or about 1 hour to about 1 week, or about 1 hour to about 2 weeks, or about 1 hour to about 3 weeks, or about 1 hour to about 4 weeks, or about 2 hours to about 6 hours, or about 2 hours to about 12 hours, or about 2 hours to about 1 day, or about 2 hours to about 2 days, or about 2 hours to about 3 days, or about 2 hours to about 4 days, or about 2 hours to about 5 days, or about 2 hours to about 6 days, or about 2 hours to about 1 week, or about 2 hours to about 2 weeks, or about 2 hours to about 3 weeks, or about 2 hours to about 4 weeks, or about 3 hours to about 12 hours, or about 3 hours to about 1 day, or about 3 hours to about 2 days, or about 3 hours to about 3 days, or about 3 hours to Approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours or approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days,Or approximately 2 to 4 days, or approximately 2 to 5 days, or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 to 4 days, or approximately 3 to 5 days, or approximately 3 to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks, or approximately 3 days to 3 weeks, or approximately 3 days to 4 weeks, or approximately 4 days to 5 days, or approximately 4 days to Approximately 6 days, or approximately 4 days to approximately 1 week, or approximately 4 days to approximately 2 weeks, or approximately 4 days to approximately 3 weeks, or approximately 4 days to approximately 4 weeks, or approximately 5 days to approximately 6 days, or approximately 5 days to approximately 1 week, or approximately 5 days to approximately 2 weeks, or approximately 5 days to approximately 3 weeks, or approximately 5 days to approximately 4 weeks, or approximately 6 days to approximately 1 week, or approximately 6 days to approximately 2 weeks, or approximately 6 days to approximately 3 weeks, or approximately 6 days to approximately 4 weeks, or approximately 1 week to approximately 2 weeks, or approximately 1 week to approximately 3 weeks, or approximately, It could be one week to about four weeks, or about two weeks to about three weeks, or about two weeks to about four weeks, or about three weeks to about four weeks.

[0108] In the method of the present invention, the organ may be the pancreas, and the method may be for the treatment of diabetes.

[0109] In the method of the present invention, the isolated cells may be neurosensory progenitor cells, and the method may be used for the treatment of retinal degenerative diseases. Retinal degenerative diseases may include age-related macular degeneration (AMD), retinitis pigmentosa (RP), retinal vasculitis, or sarcoidosis.

[0110] In some embodiments, before being implanted in a subject requiring it, the isolated graft may be washed to remove compounds represented by formulas I, II, or III.

[0111] In the embodiment, a) Before transplanting it into a subject who needs it, the isolated graft is prepared using general formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7, represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 OH, OGP', NH2, N3, NHGP', or NGP'GP" (where GP 'and GP' independently represents an alkyl group, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group.} represents a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, b) The process of implanting a graft into a subject who is in need of such treatment but has not received immunosuppressant therapy in order to suppress or prevent an immune rejection reaction to the graft. Methods for suppressing or preventing immune rejection, including the following, are disclosed.

[0112] For example, subjects who are not receiving immunosuppressant therapy and require it may be immunocompromised subjects or subjects with a weakened immune system. Examples of individuals with a weakened immune system in this embodiment include those with HIV / AIDS, cancer, and genetic disorders affecting the immune system (e.g., congenital agammaglobulinemia, congenital IgA deficiency, etc.). The risk of developing a serious illness may vary depending on the degree of immunosuppression in each individual.

[0113] According to other embodiments, General Formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 The use of a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I, is provided for use in suppressing or preventing immunorejection of an isolated graft that has been in contact with the compound before implantation into a subject requiring the compound.

[0114] For use, the compound should be applied before transplantation for approximately 1 minute to 15 minutes, or approximately 1 minute to 30 minutes, or approximately 1 minute to 1 hour, or approximately 1 minute to 6 hours, or approximately 1 minute to 12 hours, or approximately 1 minute to 24 hours, or approximately 1 minute to 48 hours, or approximately 1 minute to 72 hours, or approximately 15 minutes to 30 minutes, or approximately 15 minutes to 1 hour, or approximately 15 minutes to 6 hours, or approximately 15 minutes to 12 hours, or approximately 15 minutes to 24 hours, or approximately 15 minutes to 48 hours, or approximately 15 minutes to 72 hours, or approximately 30 minutes. or approximately 1 hour, or approximately 30 minutes to approximately 6 hours, or approximately 30 minutes to approximately 12 hours, or approximately 30 minutes to approximately 24 hours, or approximately 30 minutes to approximately 48 hours, or approximately 30 minutes to approximately 72 hours, or approximately 1 hour to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 24 hours, or approximately 1 hour to approximately 48 hours, or approximately 1 hour to approximately 72 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 24 hours, or approximately 2 hours to approximately 48 hours, or approximately 2 hours to approximately 72 hours, or approximately 6 hours to approximately 12 hours, or approximately 6 hours It can be used for intervals of up to about 24 hours, or about 6 hours to about 48 hours, or about 6 hours to about 72 hours, or about 12 hours to about 24 hours, or about 12 hours to about 48 hours, or about 12 hours to about 72 hours, or about 24 hours to about 48 hours, or about 24 hours to about 72 hours, or 1 minute, 15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, or 72 hours.

[0115] In using the present invention, subjects requiring it must undergo a period of at least approximately 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks before the implantation of the graft, or approximately 1 to approximately 2 hours, or approximately 1 to approximately 6 hours, or approximately 1 to approximately 12 hours, or approximately 1 hour to approximately 1 day, or approximately 1 hour to approximately 2 days, or approximately 1 hour to approximately 3 days, or approximately 1 hour to approximately 4 days, or approximately 1 hour to approximately 5 days, or approximately 1 hour to approximately 6 days, or approximately 1 hour to approximately 1 week, or approximately 1 hour to approximately 2 weeks, or approximately 1 Hours to approximately 3 weeks, or approximately 1 hour to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours Intervals to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 1 hour to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days, or approximately 2 days to approximately 4 days,Or approximately 2 to 5 days, or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 to 4 days, or approximately 3 to 5 days, or approximately 3 to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks, or approximately 3 days to 3 weeks, or approximately 3 days to 4 weeks, or approximately 4 to 5 days, or approximately 4 to 6 days, or approximately 4 days to 1 week, or approximately 4 days to 2 weeks, or approximately 4 days to 3 weeks, or approximately 4 days to Immunosuppressant therapy can be administered to suppress or prevent immune rejection of the graft for approximately 4 weeks, or approximately 5 to 6 days, or approximately 5 days to 1 week, or approximately 5 days to 2 weeks, or approximately 5 days to 3 weeks, or approximately 5 days to 4 weeks, or approximately 6 days to 1 week, or approximately 6 days to 2 weeks, or approximately 6 days to 3 weeks, or approximately 6 days to 4 weeks, or approximately 1 week to 2 weeks, or approximately 1 week to 3 weeks, or approximately 1 week to 4 weeks, or approximately 2 weeks to 3 weeks, or approximately 2 weeks to 4 weeks, or approximately 3 weeks to 4 weeks.

[0116] Immunosuppressant therapy may be discontinued in subjects who require it after sufficient time has elapsed for graft transplantation. The period after discontinuation of immunosuppressant therapy, in other words, sufficient time for transplantation, is at least about 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks after graft transplantation, or about 1 hour to about 2 hours, or about 1 hour. Up to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 1 day, or approximately 1 hour to approximately 2 days, or approximately 1 hour to approximately 3 days, or approximately 1 hour to approximately 4 days, or approximately 1 hour to approximately 5 days, or approximately 1 hour to approximately 6 days, or approximately 1 hour to approximately 1 week, or approximately 1 hour to approximately 2 weeks, or approximately 1 hour to approximately 3 weeks, or approximately 1 hour to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours Up to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours or approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days Between, or approximately 1 to 3 days, or approximately 1 to 4 days, or approximately 1 to 5 days, or approximately 1 to 6 days, or approximately 1 day to 1 week, or approximately 1 day to 2 weeks, or approximately 1 day to 3 weeks, or approximately 1 day to 4 weeks, or approximately 2 to 3 days, or approximately 2 to 4 days, or approximately 2 to 5 days, or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 to 4 days, or approximately 3 to 5 days, or approximately 3 to 6 days,Alternatively, it could be approximately 3 days to approximately 1 week, or approximately 3 days to approximately 2 weeks, or approximately 3 days to approximately 3 weeks, or approximately 3 days to approximately 4 weeks, or approximately 4 days to approximately 5 days, or approximately 4 days to approximately 6 days, or approximately 4 days to approximately 1 week, or approximately 4 days to approximately 2 weeks, or approximately 4 days to approximately 3 weeks, or approximately 4 days to approximately 4 weeks, or approximately 5 days to approximately 6 days, or approximately 5 days to approximately 1 week, or approximately 5 days to approximately 2 weeks, or approximately 5 days to approximately 3 weeks, or approximately 5 days to approximately 4 weeks, or approximately 6 days to approximately 1 week, or approximately 6 days to approximately 2 weeks, or approximately 6 days to approximately 3 weeks, or approximately 6 days to approximately 4 weeks, or approximately 1 week to approximately 2 weeks, or approximately 1 week to approximately 3 weeks, or approximately 1 week to approximately 4 weeks, or approximately 2 weeks to approximately 3 weeks, or approximately 2 weeks to approximately 4 weeks, or approximately 3 weeks to approximately 4 weeks.

[0117] According to other embodiments, the subjects requiring this may be those who have not received immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0118] For example, subjects who are not receiving immunosuppressant therapy and require it may be immunocompromised subjects or subjects with a weakened immune system. Examples of individuals with a weakened immune system in this embodiment include those with HIV / AIDS, cancer, and genetic disorders affecting the immune system (e.g., congenital agammaglobulinemia, congenital IgA deficiency, etc.). The risk of developing a serious illness may vary depending on the degree of immunosuppression in each individual.

[0119] According to another embodiment, a general formula I: is used for suppressing or preventing immune rejection of an isolated graft that has come into contact with the compound before implantation into a subject requiring it. [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R 2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R 1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 The following are provided: a gem-difluorinated C-glycopeptide compound represented by}, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by general formula I with a base or acid.

[0120] For use, the compound should be left on the skin for at least approximately 1 minute, or approximately 15 minutes, or approximately 30 minutes, or approximately 1 hour, or approximately 6 hours, or approximately 12 hours, or approximately 24 hours, or approximately 48 hours, or approximately 72 hours, or approximately 1 minute to approximately 15 minutes, or approximately 1 minute to approximately 30 minutes, or approximately 1 minute to approximately 1 hour, or approximately 1 minute to approximately 6 hours, or approximately 1 minute to approximately 12 hours, or approximately 1 minute to approximately 24 hours, or approximately 1 minute to approximately 48 hours, or approximately 1 minute to approximately 72 hours, or approximately 15 minutes to approximately 30 minutes, or approximately 15 minutes to approximately 1 hour, or approximately 15 minutes to approximately 6 hours, or approximately 15 minutes to approximately 12 hours, or approximately 15 minutes to approximately 24 hours before transplantation. , or approximately 15 minutes to approximately 48 hours, or approximately 15 minutes to approximately 72 hours, or approximately 30 minutes to approximately 1 hour, or approximately 30 minutes to approximately 6 hours, or approximately 30 minutes to approximately 12 hours, or approximately 30 minutes to approximately 24 hours, or approximately 30 minutes to approximately 48 hours, or approximately 30 minutes to approximately 72 hours, or approximately 1 hour to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 24 hours, or approximately 1 hour to approximately 48 hours, or approximately 1 hour to approximately 72 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 24 hours, or approximately 2 hours to approximately 48 hours, or approximately 2 hours to approximately 72 hours, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 2 It can be used for 4 hours, or approximately 6 to 48 hours, or approximately 6 to 72 hours, or approximately 12 to 24 hours, or approximately 12 to 48 hours, or approximately 12 to 72 hours, or approximately 24 to 48 hours, or approximately 24 to 72 hours, or approximately 1 minute, 15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, or 72 hours.

[0121] Subjects requiring this treatment should undergo a period of at least approximately 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks before graft implantation, or approximately 1 to 2 hours, or approximately 1 to 6 hours, or approximately 1 to 12 hours, or approximately 1 hour to 1 day, or approximately 1 hour to 2 days, or approximately 1 hour to 3 days, or approximately 1 hour to 4 days, or approximately 1 hour to 5 days, or approximately 1 hour to 6 days, or approximately 1 hour to 1 week, or approximately 1 hour to 2 weeks, or approximately 1 hour to 3 weeks, or approximately 1 hour Up to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours or approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 1 2 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days, or approximately 2 days to approximately 4 days, or approximately 2 days to approximately 5 days,Or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 days to 4 days, or approximately 3 days to 5 days, or approximately 3 days to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks, or approximately 3 days to 3 weeks, or approximately 3 days to 4 weeks, or approximately 4 days to 5 days, or approximately 4 days to 6 days, or approximately 4 days to 1 week, or approximately 4 days to 2 weeks, or approximately 4 days to 3 weeks, or approximately 4 days to 4 weeks, or They may receive immunosuppressant therapy to suppress or prevent immune rejection of the graft for approximately 5 to 6 days, or approximately 5 days to 1 week, or approximately 5 days to 2 weeks, or approximately 5 days to 3 weeks, or approximately 5 days to 4 weeks, or approximately 6 days to 1 week, or approximately 6 days to 2 weeks, or approximately 6 days to 3 weeks, or approximately 6 days to 4 weeks, or approximately 1 week to 2 weeks, or approximately 1 week to 3 weeks, or approximately 1 week to 4 weeks, or approximately 2 weeks to 3 weeks, or approximately 2 weeks to 4 weeks, or approximately 3 weeks to 4 weeks.

[0122] Immunosuppressant therapy may be discontinued in subjects requiring it after sufficient time has elapsed for graft transplantation. The period after discontinuation of immunosuppressant therapy, in other words, sufficient time for transplantation, is from at least 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks after graft transplantation, or about 1 to 2 hours, or about 1 to 6 hours, or about 1 to 12 hours, or about 1 hour to 1 day, or about 1 hour to 2 days, or about 1 hour to 3 days, or about 1 hour Up to approximately 4 days, or approximately 1 hour to approximately 5 days, or approximately 1 hour to approximately 6 days, or approximately 1 hour to approximately 1 week, or approximately 1 hour to approximately 2 weeks, or approximately 1 hour to approximately 3 weeks, or approximately 1 hour to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours Up to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours Up to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week Between, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days, or approximately 2 days to approximately 4 days, or approximately 2 days to approximately 5 days, or approximately 2 days to approximately 6 days, or approximately 2 days to approximately 1 week, or approximately 2 days to approximately 2 weeks, or approximately 2 days to approximately 3 weeks, or approximately 2 days to approximately 4 weeks, or approximately 3 days to approximately 4 days, or approximately 3 days to approximately 5 days, or approximately 3 days to approximately 6 days, or approximately 3 days to approximately 1 week, or approximately 3 days to approximately 2 weeks, or approximately 3 days to approximately 3 weeks, or approximately 3 days to approximately 4 weeks, or approximately 4 days to approximately 5 days,Alternatively, it could be approximately 4 to 6 days, or approximately 4 days to 1 week, or approximately 4 days to 2 weeks, or approximately 4 days to 3 weeks, or approximately 4 days to 4 weeks, or approximately 5 to 6 days, or approximately 5 days to 1 week, or approximately 5 days to 2 weeks, or approximately 5 days to 3 weeks, or approximately 5 days to 4 weeks, or approximately 6 days to 1 week, or approximately 6 days to 2 weeks, or approximately 6 days to 3 weeks, or approximately 6 days to 4 weeks, or approximately 1 week to 2 weeks, or approximately 1 week to 3 weeks, or approximately 1 week to 4 weeks, or approximately 2 weeks to 3 weeks, or approximately 2 weeks to 4 weeks, or approximately 3 weeks to 4 weeks.

[0123] According to other embodiments, the subjects requiring this may be subjects who have not received immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0124] For example, subjects who are not receiving immunosuppressant therapy and require it may be immunocompromised subjects or subjects with a weakened immune system. Examples of individuals with a weakened immune system in this embodiment include those with HIV / AIDS, cancer, and genetic disorders affecting the immune system (e.g., congenital agammaglobulinemia, congenital IgA deficiency, etc.). The risk of developing a serious illness may vary depending on the degree of immunosuppression in each individual.

[0125] According to other embodiments, in order to suppress or prevent immune rejection of an isolated graft that has come into contact with the compound, before implanting it into a subject requiring it, the general formula I: [ka] {In the formula, N is an integer from 1 to 5. R 4 This represents H, AA1, or AA1-AA2, R 5 represents OH, AA1, or AA1-AA2, AA1 and AA2 independently represent amino acids with nonpolar side chains, and R 1 , R2 , R 3 It is an independent group, Here R 1 , R 2 and R 3 Two of them are H and CH 3、 CH2Ph, CH(CH3) 2、 Selected from CH2CH(CH3)2 or CH(CH3)CH2CH3, the remaining R 1 , R 2 , R 3 teeth [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H, or a free or protected alcohol functional group. ] represents, and, R1 and R 2 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 3 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 1 and R 3 However, when representing H, CH3, CH2Ph, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 2 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. R 2 and R 3 However, when representing H, CH3, CH(CH3)2, CH2CH(CH3)2, or CH(CH3)CH2CH3, R 1 teeth, [ka] [In the formula, n is an integer between 3 and 4. Y and Y' are independent groups. Here, Y and Y' represent H, OR, N3, NR'R'' or SR'''', Here, R represents H, a benzyl group, a trimethylsilyl group, a tert-butyldimethylsilyl group, a tert-butyldiphenylsilyl group, or an acetate group. R' and R'' independently represent H, alkyl group, allyl group, benzyl group, tosylate group, C(=O)-alkyl group, or C(=O)-Bn. R''' represents H, an alkyl group, or an acetate group. R 6 This represents H, CH3, CH2OH, CH2-glycoside group, or CH2-OGP (where GP represents a protecting group selected from alkyl group, benzyl group, trimethylsilyl group, tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group, or acetate group). R 7 , represents OH, OGP', NH2, N3, NHGP', or NGP'GP'' (where GP' and GP'' are independently selected from alkyl groups, benzyl groups, trimethylsilyl groups, tert-butyldimethylsilyl groups, tert-butyldiphenylsilyl groups, or acetate groups). R 8 represents a hydrogen atom H or a free or protected alcohol functional group. Disclosed are gem-difluorinated C-glycopeptide compounds represented by}, or isolated grafts in contact with pharmaceutically acceptable addition salts, hydrates, or solvates of the compound represented by general formula I.

[0126] In use, the compound should be applied before transplantation for at least approximately 1 minute, or approximately 15 minutes, or approximately 30 minutes, or approximately 1 hour, or approximately 6 hours, or approximately 12 hours, or approximately 24 hours, or approximately 48 hours, or approximately 72 hours, or approximately 1 minute to approximately 15 minutes, or approximately 1 minute to approximately 30 minutes, or approximately 1 minute to approximately 1 hour, or approximately 1 minute to approximately 6 hours, or approximately 1 minute to approximately 12 hours, or approximately 1 minute to approximately 24 hours, or approximately 1 minute to approximately 48 hours, or approximately 1 minute to approximately 72 hours, or approximately 15 minutes to approximately 30 minutes, or approximately 15 minutes to approximately 1 hour, or approximately 15 minutes to approximately 6 hours, or approximately 15 minutes to Approximately 12 hours, or approximately 15 minutes to approximately 24 hours, or approximately 15 minutes to approximately 48 hours, or approximately 15 minutes to approximately 72 hours, or approximately 30 minutes to approximately 1 hour, or approximately 30 minutes to approximately 6 hours, or approximately 30 minutes to approximately 12 hours, or approximately 30 minutes to approximately 24 hours, or approximately 30 minutes to approximately 48 hours, or approximately 30 minutes to approximately 72 hours, or approximately 1 hour to approximately 6 hours, or approximately 1 hour to approximately 12 hours, or approximately 1 hour to approximately 24 hours, or approximately 1 hour to approximately 48 hours, or approximately 1 hour to approximately 72 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 24 hours, or approximately 2 hours to approximately 48 hours It can be used for approximately 2 hours to approximately 72 hours, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 24 hours, or approximately 6 hours to approximately 48 hours, or approximately 6 hours to approximately 72 hours, or approximately 12 hours to approximately 24 hours, or approximately 12 hours to approximately 48 hours, or approximately 12 hours to approximately 72 hours, or approximately 24 hours to approximately 48 hours, or approximately 24 hours to approximately 72 hours, or approximately 1 minute, 15 minutes, 30 minutes, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, or 72 hours.

[0127] Subjects requiring this treatment should undergo a period of at least approximately 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks before graft implantation, or approximately 1 to 2 hours, or approximately 1 to 6 hours, or approximately 1 to 12 hours, or approximately 1 hour to 1 day, or approximately 1 hour to 2 days, or approximately 1 hour to 3 days, or approximately 1 hour to 4 days, or approximately 1 hour to 5 days, or approximately 1 hour to 6 days, or approximately 1 hour to 1 week, or approximately 1 hour to 2 weeks, or approximately 1 hour to 3 weeks, or approximately 1 hour Up to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours or approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 1 2 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 day to approximately 5 days, or approximately 1 day to approximately 6 days, or approximately 1 day to approximately 1 week, or approximately 1 day to approximately 2 weeks, or approximately 1 day to approximately 3 weeks, or approximately 1 day to approximately 4 weeks, or approximately 2 days to approximately 3 days, or approximately 2 days to approximately 4 days, or approximately 2 days to approximately 5 days,Or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 days to 4 days, or approximately 3 days to 5 days, or approximately 3 days to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks, or approximately 3 days to 3 weeks, or approximately 3 days to 4 weeks, or approximately 4 days to 5 days, or approximately 4 days to 6 days, or approximately 4 days to 1 week, or approximately 4 days to 2 weeks, or approximately 4 days to 3 weeks, or approximately 4 days to 4 weeks, or They may receive immunosuppressant therapy to suppress or prevent immune rejection of the graft for approximately 5 to 6 days, or approximately 5 days to 1 week, or approximately 5 days to 2 weeks, or approximately 5 days to 3 weeks, or approximately 5 days to 4 weeks, or approximately 6 days to 1 week, or approximately 6 days to 2 weeks, or approximately 6 days to 3 weeks, or approximately 6 days to 4 weeks, or approximately 1 week to 2 weeks, or approximately 1 week to 3 weeks, or approximately 1 week to 4 weeks, or approximately 2 weeks to 3 weeks, or approximately 2 weeks to 4 weeks, or approximately 3 weeks to 4 weeks.

[0128] Immunosuppressant therapy may be discontinued after sufficient time has elapsed to allow for graft implantation in subjects requiring it. The period after discontinuation of immunosuppressant therapy, in other words, sufficient time for implantation, is at least about 1 hour, 2 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 4 weeks after graft implantation, or about 1 to 2 hours, or about 1 to 6 hours, or about 1 to 12 hours. Or approximately 1 hour to approximately 1 day, or approximately 1 hour to approximately 2 days, or approximately 1 hour to approximately 3 days, or approximately 1 hour to approximately 4 days, or approximately 1 hour to approximately 5 days, or approximately 1 hour to approximately 6 days, or approximately 1 hour to approximately 1 week, or approximately 1 hour to approximately 2 weeks, or approximately 1 hour to approximately 3 weeks, or approximately 1 hour to approximately 4 weeks, or approximately 2 hours to approximately 6 hours, or approximately 2 hours to approximately 12 hours, or approximately 2 hours to approximately 1 day, or approximately 2 hours to approximately 2 days, or approximately 2 hours to approximately 3 days, or approximately 2 hours to approximately 4 days, or approximately 2 hours to approximately 5 days, or approximately 2 hours to approximately 6 days, or approximately 2 hours to approximately 1 week, or Approximately 2 hours to approximately 2 weeks, or approximately 2 hours to approximately 3 weeks, or approximately 2 hours to approximately 4 weeks, or approximately 3 hours to approximately 12 hours, or approximately 3 hours to approximately 1 day, or approximately 3 hours to approximately 2 days, or approximately 3 hours to approximately 3 days, or approximately 3 hours to approximately 4 days, or approximately 3 hours to approximately 5 days, or approximately 3 hours to approximately 6 days, or approximately 3 hours to approximately 1 week, or approximately 3 hours to approximately 2 weeks, or approximately 3 hours to approximately 3 weeks, or approximately 3 hours to approximately 4 weeks, or approximately 6 hours to approximately 12 hours, or approximately 6 hours to approximately 1 day, or approximately 6 hours to approximately 2 days, or approximately 6 hours to approximately 3 days, or approximately 6 hours to approximately 4 days, or approximately 6 hours to approximately 5 days, or approximately 6 hours to approximately 6 days, or approximately 6 hours to approximately 1 week, or approximately 6 hours to approximately 2 weeks, or approximately 6 hours to approximately 3 weeks, or approximately 6 hours to approximately 4 weeks, or approximately 12 hours to approximately 1 day, or approximately 12 hours to approximately 2 days, or approximately 12 hours to approximately 3 days, or approximately 12 hours to approximately 4 days, or approximately 12 hours to approximately 5 days, or approximately 12 hours to approximately 6 days, or approximately 12 hours to approximately 1 week, or approximately 12 hours to approximately 2 weeks, or approximately 12 hours to approximately 3 weeks, or approximately 12 hours to approximately 4 weeks, or approximately 1 day to approximately 2 days, or approximately 1 day to approximately 3 days, or approximately 1 day to approximately 4 days, or approximately 1 to 5 days, or approximately 1 to 6 days, or approximately 1 day to 1 week, or approximately 1 day to 2 weeks, or approximately 1 day to 3 weeks, or approximately 1 day to 4 weeks, or approximately 2 to 3 days, or approximately 2 to 4 days, or approximately 2 to 5 days, or approximately 2 to 6 days, or approximately 2 days to 1 week, or approximately 2 days to 2 weeks, or approximately 2 days to 3 weeks, or approximately 2 days to 4 weeks, or approximately 3 to 4 days, or approximately 3 to 5 days, or approximately 3 to 6 days, or approximately 3 days to 1 week, or approximately 3 days to 2 weeks,Alternatively, it could be approximately 3 days to approximately 3 weeks, or approximately 3 days to approximately 4 weeks, or approximately 4 days to approximately 5 days, or approximately 4 days to approximately 6 days, or approximately 4 days to approximately 1 week, or approximately 4 days to approximately 2 weeks, or approximately 4 days to approximately 3 weeks, or approximately 4 days to approximately 4 weeks, or approximately 5 days to approximately 6 days, or approximately 5 days to approximately 1 week, or approximately 5 days to approximately 2 weeks, or approximately 5 days to approximately 3 weeks, or approximately 5 days to approximately 4 weeks, or approximately 6 days to approximately 1 week, or approximately 6 days to approximately 2 weeks, or approximately 6 days to approximately 3 weeks, or approximately 6 days to approximately 4 weeks, or approximately 1 week to approximately 2 weeks, or approximately 1 week to approximately 3 weeks, or approximately 1 week to approximately 4 weeks, or approximately 2 weeks to approximately 3 weeks, or approximately 2 weeks to approximately 4 weeks, or approximately 3 weeks to approximately 4 weeks.

[0129] According to other embodiments, the subjects requiring this may be subjects who have not received immunosuppressant therapy to suppress or prevent immune rejection of the graft.

[0130] For example, subjects who are not receiving immunosuppressant therapy and require it may be immunocompromised subjects or subjects with a weakened immune system. Examples of individuals with a weakened immune system in this embodiment include those with HIV / AIDS, cancer, and genetic disorders affecting the immune system (e.g., congenital agammaglobulinemia, congenital IgA deficiency, etc.). The risk of developing a serious illness may vary depending on the degree of immunosuppression in each individual.

[0131] The present invention comprises compounds as shown, and (if possible) also comprises individual diastereomers, enantiomers, and epimers of the compounds, as well as mixtures of those diastereomers and / or enantiomers, including racemic mixtures. While the specific stereochemistry disclosed herein is preferred, other stereoisomers, including diastereomers, enantiomers, epimers, and mixtures thereof, may also be useful. Inactive or low-activity diastereomers and enantiomers are useful for scientific studies related to target and / or activation mechanisms.

[0132] The compounds disclosed herein may be used in pharmaceutical compositions comprising (a) the compound or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier. The compounds may be used in pharmaceutical compositions comprising one or more other pharmaceutically active ingredients. Furthermore, the compounds may be used in pharmaceutical compositions in which the compound represented by formula I, II, or III or a pharmaceutically acceptable salt thereof is the sole active ingredient.

[0133] Compounds represented by structural formulas I, II, and / or III may contain one or more asymmetric centers and can therefore exist as racemates and racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers. The present invention is intended to encompass all such isomers of compounds represented by structural formulas I, II, and / or III.

[0134] Compounds represented by structural formulas I, II, and / or III can be separated into their individual diastereoisomers by fractional crystallization from a suitable solvent such as methanol, ethyl acetate, or a mixture thereof, or by chiral chromatography using an optically active stationary phase. Absolute stereochemistry can be determined, if necessary, by X-ray crystallography of crystalline products or intermediates derivatized using reagents containing asymmetric centers of known absolute configurations.

[0135] Alternatively, any stereoisomer of the compounds of general structural formula I, structural formula II, and / or structural formula III can be obtained by stereospecific synthesis using optically pure starting materials or reagents with known absolute configurations.

[0136] If desired, the racemic mixture of compounds may be separated so that the individual enantiomers can be isolated. This separation can be carried out by methods well known in the art, such as coupling the racemic mixture of compounds with an enantiomerically pure compound to form a diastereomer mixture, and then separating the individual diastereomers by standard methods such as fractionation crystallization or chromatography. The coupling reaction is often the formation of a salt using an enantiomerically pure acid or base. Subsequently, the diastereomer derivative can be converted back to the pure enantiomer by cleavage of an added chiral residue. The racemic mixture of compounds can also be separated directly by chromatography using a chiral stationary phase, a method well known in the art.

[0137] Some of the compounds described herein contain olefinic double bonds, and unless otherwise specified, this means that both E and Z geometric isomers are included.

[0138] Some of the compounds described herein may exist as tautomers having different hydrogen attachment sites with one or more double bond shifts. For example, ketones and their enol forms are keto-enol tautomers. Individual tautomers and mixtures thereof are encompassed by the compounds of the present invention.

[0139] In compounds represented by general formulas I, II, and / or III, atoms may exhibit natural isotopic abundances, or one or more atoms may be artificially enriched with specific isotopes having the same atomic number but different atomic weights or mass numbers from those abundant in nature. The present invention is intended to encompass all appropriate isotopic variations of compounds represented by general formulas I, II, and / or III. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H). Protium is the most common hydrogen isotope found in nature. Enrichment with deuterium may provide therapeutic benefits such as increased in vivo half-life or reduced required doses, and may also provide compounds useful as criteria for characterizing biological samples. Isotope-enriched compounds can be prepared without excessive experimentation by conventional techniques well known to those skilled in the art.

[0140] • Salts and preparations As used herein, references to compounds represented by structural formulas I, II, and / or III will be understood to include pharmaceutically acceptable salts, as well as pharmaceutically unacceptable salts when used as precursors to the free compound or its pharmaceutically acceptable salts, or in other synthetic operations. The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable, non-toxic base or acid, including inorganic or organic bases and inorganic or organic acids. Salts of basic compounds encompassed by the term "pharmaceutically acceptable salt" refer to non-toxic salts of the compounds of the present invention, which are generally prepared by reacting a free base with a suitable organic or inorganic acid. Representative salts of the basic compounds of the present invention include, but are not limited to, the following: acetate, benzenesulfonate, benzoate, bicarbonate, bissulfate, vitrate, borate, bromide, camsilate, carbonate, chloride, clavulanate, citrate, edetrate, edisylate, estrate, esylate, fumarate, gluceptate, gluconate, glutamate, hexylresorcinate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate. Lactate, lactobionate, laurate, malatate, maleate, mandelate, mesylate, methyl bromide, methyl nitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, theocrate, tosylate, triethiodide, and valerate. Furthermore, when the compounds of the present invention support an acidic moiety, suitable pharmaceutically acceptable salts include, but are not limited to, salts derived from inorganic bases such as aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganese, manganese triphosphate, potassium, sodium, and zinc. Particularly preferred are ammonium salts, calcium salts, magnesium salts, potassium salts, and sodium salts.Examples of pharmaceutically acceptable salts derived from organic non-toxic bases include salts of primary, secondary, and tertiary amines, cyclic amines, and basic ion exchange resins, such as arginine, β-in, caffeine, choline, N,N-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, tripropylamine, and tromethamine.

[0141] Furthermore, if the compound of the present invention contains a carboxylic acid (-COOH) or alcohol group, carboxylic acid derivatives such as a methyl group, an ethyl group, or a pivaloyloxymethyl group, or pharmaceutically acceptable esters of acetyl groups, pivaloyl groups, benzoyl groups, and acyl derivatives of alcohols such as aminoacyls can be used. These include ester groups and acyl groups known in the art for modifying solubility or hydrolysis for use as a sustained-release or prodrug formulation.

[0142] Solvates, particularly hydrates, of compounds represented by structural formulas I, II, and / or III are also included in the present invention.

[0143] According to the embodiment, the compounds represented by structural formulas I, II, and / or III may be included in various formulations for use as pharmaceuticals.

[0144] The aqueous suspension contains the active substance when miscible with an excipient suitable for the preparation of the aqueous suspension. Such excipients may be suspending agents, dispersing agents or wetting agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum and acacia gum, or naturally occurring phosphatides such as lecithin, or condensates of alkylene oxides and fatty acids such as polyoxyethylene stearate, or condensates of ethylene oxide and long-chain aliphatic alcohols such as heptadecaethylene-oxycetanol, or condensation products of ethylene oxide with fatty acids and partial ester derivatives from hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with fatty acids and partial ester derivatives from hexitol anhydrides such as polyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, such as ethyl, n-propyl, or p-hydroxybenzoate; one or more colorants; one or more flavorings; and one or more sweeteners, such as sucrose, saccharin, or aspartame.

[0145] Oily suspensions can be formulated by suspending the active ingredient in a vegetable oil such as arachis oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain thickeners such as beeswax, hard paraffin, or cetyl alcohol. The aforementioned sweeteners and flavorings may be added to provide a palatable oral formulation. These compositions can be preserved by the addition of antioxidants such as ascorbic acid.

[0146] Dispersible powders and granules suitable for preparing aqueous suspensions by dispersion of water provide an active ingredient mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are exemplified by those already described above. Additional excipients such as sweeteners, flavorings, and colorants may also be present.

[0147] Furthermore, the pharmaceutical composition of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be, for example, a vegetable oil such as olive oil or arachis oil, or a mineral oil such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be, for example, naturally occurring phospholipids such as soybeans and lecithin, and esters or partial esters obtained from fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of ethylene oxide such as polyoxyethylene sorbitan monooleate and the partial ester. The emulsion may also contain sweeteners and flavorings.

[0148] The pharmaceutical composition may be in the form of a sterile, injectable aqueous or oily suspension. This suspension may be formulated according to known techniques using the appropriate dispersants or wetting agents and suspending agents described above. The sterile injectable formulation may also be a sterile injectable solution or suspension in a non-toxic, parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable vehicles and solvents that may be employed include water, Ringer's solution, and isotonic sodium chloride solution. Furthermore, sterile fixatives have conventionally been employed as solvents or suspension media. For this purpose, any sterile fixative, including synthetic mono or diglycerides, may be employed. Additionally, fatty acids such as oleic acid are used in the preparation of injectable formulations.

[0149] According to the embodiments, cells are isolated using methods known in the art for their preparation. For example, cells can be isolated from a donor using a mixture of enzymes such as collagenase I and collagenase II, thermolysin, non-Clostridium neutral proteases, or other enzymes used for such purposes. The isolated cells can be cultured in a standard tissue culture flask under normal tissue culture conditions.

[0150] According to one embodiment, the neuronal sensory progenitor cells are approximately 0.01 mg / mL to approximately 5 mg / mL, or approximately 0.1 mg / mL to approximately 5 mg / mL, or approximately 0.5 mg / mL to approximately 5 mg / mL, or approximately 1 mg / mL to approximately 5 mg / mL, or approximately 3 mg / mL to approximately 5 mg / mL, or approximately 0.01 mg / mL to approximately 3 mg / mL, or approximately 0.1 mg / mL to approximately 3 mg / mL, or approximately 0.5 mg / mL to approximately 3 mg / mL, or approximately 1 mg / mL to approximately 3 mg / mL, or approximately 0.01 mg / mL The patient may be treated with a gem-difluorinated C-glycopeptide compound represented by general formula I, preferably a compound represented by formula II, and more preferably a compound represented by formula III, at concentrations ranging from approximately 1 mg / mL, or approximately 0.1 mg / mL to approximately 1 mg / mL, or approximately 0.5 mg / mL to approximately 1 mg / mL, or approximately 0.01 mg / mL to approximately 0.5 mg / mL, or approximately 0.01 mg / mL to approximately 0.1 mg / mL, or approximately 3 mg / mL. According to embodiments, the above amounts are considered to be therapeutically effective amounts for the purposes of the present invention.

[0151] According to other embodiments, the gem-difluorolated C-glycopeptide compound is brought into contact with the cells for a sufficient time to improve cell viability and survival rate. According to embodiments, a sufficient time may be about 12 to 120 hours, or about 12 to about 96 hours, or about 12 to about 72 hours, or about 12 to about 48 hours, or about 12 to about 24 hours, or about 120 hours, or about 96 hours, or about 72 hours, or about 48 hours, or about 24 hours, or about 12 hours, or about 10 hours, or about 8 hours, or about 6 hours, or about 4 hours, or about 2 hours, or about 1 hour. According to the embodiment, the neurosensory progenitor cells isolated herein are exposed to the compound for 1 hour, 55 minutes, 50 minutes, 45 minutes, 40 minutes, 35 minutes, 30 minutes, 25 minutes, 20 minutes, 15 minutes, 10 minutes, 5 minutes, 4 minutes, 3 minutes, 2 minutes, 1 minute, 45 seconds, or 30 seconds, or at least 1 hour, or at least 55 minutes, or at least 50 minutes, or at least 45 minutes, or at least 40 minutes, or at least 35 minutes, or at least 30 minutes, or at least 25 minutes, or at least 20 minutes, or at least 15 minutes, or at least 10 minutes, or at least 5 minutes, or at least 4 minutes, or at least 3 minutes, or at least 2 minutes, or at least 1 minute, or at least 45 seconds, or at least 30 seconds.

[0152] In another embodiment, a cell preparation prepared according to the method of the present invention in a pharmaceutically acceptable carrier is disclosed. According to one embodiment, the cell preparation may be used for the preparation of a pharmaceutical for cell transplantation. According to another embodiment, the cell preparation may be used for cell transplantation.

[0153] In another embodiment, a transplantation method is disclosed, which includes transplanting a cell preparation of the present invention into a subject requiring it. The subject may be a mammal, preferably a human.

[0154] The present invention will be more readily understood by referring to the following embodiments, which are given not to limit its scope, but to illustrate the invention.

[0155] Example 1 The purpose of this study is to investigate the long-term (6-month) survival and functional activity of photoreceptor progenitor cells (PPCs) subretinally transplanted into the eyes of rabbits with retinal degeneration, using anti-aging glycoprotein (AAGP). TM The objective is to clarify the effect of 24-hour pretreatment using 4 mg / mL. AAGP TM Anti-aging glycoprotein DMEM / F12: Dulbecco's modified Eagle medium / nutrient mixture F12 EB: Embryoid body ERG: Electroretinal Potential GCL: Ganglion cell layer hESC: Human Embryonic Stem Cells ID: Identification IGF-1: Insulin-like growth factor 1 INL: inner nuclear layer ONL: outer nuclear layer PBS: Phosphate-buffered saline PPCs: Photoreceptor progenitor cells RD: Retinal degeneration

[0156] InVivo System Species: Rabbit (European rabbit) Strain; White Albino New Zealand, Crl:KBL(NZW)BR Source: Charles River Canada Ltd. 324 Cent Regis nodes, CP400 St. Constant, Quebec, Canada Sex: Female Age: 5 weeks old (at the time of arrival at the testing facility) Weight: 1.25-1.50 kg (at the time of arrival at the testing facility)

[0157] Nine New Zealand rabbits were obtained from Charles River Ltd. at 5 weeks of age and weighing 1.25–1.50 kg. The animals were maintained on a 12-hour light-dark cycle, and the study was conducted in accordance with protocols compliant with the Board of Animal Monitoring in Canada, with the approval of the Animal Care Committee at the University of British Columbia and accompanied by a statement from the Society for Visual and Ophthalmic Research on the Use of Animals in Visual Research.

[0158] Species identification and acclimatization The animals were allowed to acclimate to the laboratory environment for at least five days before being placed in different research groups. Each animal was uniquely identified by having an identification (ID) number attached to its ear using a permanent marker.

[0159] Experimental Design PPCs were prepared in vitro from pluripotent human embryonic stem cells (hESCs), and then administered in either culture medium only (control group) or 4 mg / mL AAGP. TM The cells were incubated in the culture medium for 24 hours. After incubation, treated and untreated PPCs were labeled and injected into the subretinal space of immunosuppressed rabbits with induced retinal degeneration (4-5 rabbits / group). The animals were euthanized after 6 months, the retinas were harvested, and cell viability was determined on excised retinal flat mounts by imaging using a confocal laser scanning system. The functional activity of the transplanted cells was evaluated by immunostaining for synaptic markers (Ribeye and Bassoon) and photoreceptor markers (Arrestin and Blue Opsin). A summary of the research plan is shown in the table and flowchart in Figure 1. [Table 1]

[0160] procedure Induction of retinal degeneration Retinal degeneration was induced on day 1 of the study. Animals were given 20 mg / kg of ketamine (Narketan). TM, 100 mg / mL; Betquinol N.-A. Incorporated, Lavaltrie, Quebec, Canada) and 5 mg / kg of xalazine (Rompun) TM The subjects were placed under general anesthesia by intramuscular administration of 20 mg / mL (Bayer Incorporated, Toronto, Canada), and their pupils were dilated with 1% tropicamide (Bosch & Lomb, Rochester, New York, USA). Povidone-iodine (Alcon) antimicrobial solution was used to cleanse the eyeballs, eyelids, surrounding skin, and the subjects' fur. Tear-gel (Alcon) was applied to the cornea during surgery.

[0161] Phosphate-buffered saline (PBS) (Life Technologies, Inc., Toronto, Canada) was collected in a 1 mL syringe connected to a 30 G needle (BD, Canada). 250 μL of PBS was slowly injected subretinically to form a uniform blister clearly visible under a surgical microscope. Care was taken to keep the tip of the syringe within the blister during injection to minimize backflow. Retinal degeneration was induced in one eye of each animal, while the other eye remained intact.

[0162] Postoperative topical antibiotics (1% chloramphenicol ophthalmic ointment, Netcare Healthcare UK Limited, London) were administered. Postoperative follow-up showed no signs of extraocular or intraocular infection or inflammation in either eye.

[0163] Immunosuppressants Transplant recipients take the immunosuppressant cyclosporine A (Neoral) for one week before transplantation and two weeks after transplantation. TM The patient received 10 mg / kg / day of cyclosporine A (Novartis, East Hanover, New Zealand, USA) orally daily, and then discontinued administration. Cyclosporine A was administered orally as garbage.

[0164] Preparation of PPCs Photoreceptor progenitor cells (PPCs) were induced from the pluripotent human ES cell (hESC) line WA09 (Wisell) according to the protocols of Yanai et al. 2013 and Yanai et al. 2015.

[0165] 1) Complete TeSR in a humidity provided by 37°C, 5% CO2, and distilled water in a tray at the bottom of the incubator. TM Cells were cultured in two culture media (SteamCell Technologies) [1, 2]. Cells were cultured in 6-well plates coated with growth factor-reduced Matrigel (BD Biosciences) (Nunclonδ surface, ThermoScientific).

[0166] hESC was differentiated into PPCs by a multi-step procedure as shown in the figure below, which is described in detail in Yanai et al. 2013 and Yanai et al. 2015 and in the following sections.

[0167] 2) To produce size-controlled embryoid bodies (EBs), hESCs are washed with DMEM / F12 (Life Technology Co., Ltd.) and Accutase TM (Stem Cell Technologies) Cells were dissociated into single cells using 0.75 mL / well. The cells were incubated at 37°C for 6-10 minutes until most cells were detached, then harvested in 4 mL / well of DMEM / F12, centrifuged at 300 g for 5 minutes, and the pellet was fed into EB forming medium [TeSR with 10 μM Rho-related coil kinase (ROCK) inhibitor (Stem Cell Technologies)]. TM The cells were resuspended in [2]. The viable cells were counted using trypan blue, and then... AggreWell400 TM Each well of the plate (Stem Cell Technologies) contains 1.2 × 10 6 Numerous surviving hESCs were seeded. The plate was centrifuged with 100g for 3 minutes and left to stand in an incubator β at 37°C for 24 hours.

[0168] 3) After incubation, EBs were gently pipetted through a 37 μm cell strainer (Stem Cell Technologies), unintegrated single cells were discarded, and the cells were distributed into ultra-low adhesion 24-well dishes (Corning). For the next 3 days, the EBs were maintained in EB resuspension, also known as retinal induction medium [(DMEM / F12, 10% knockout serum substitution, 2% custom B-27 and 1% N-2 supplement (Life Technologies), 1 ng / mL recombinant human DKK-1, 1 ng / mL mouse noggin and 5 ng / mL recombinant human insulin-like growth factor (IGF-1) (R&D Systems)].

[0169] 4) On day 4, the EBs were collected in a 50 ml tube, centrifuged at 100 g for 5 minutes, resuspended in PPC differentiation medium [DMEM / F12, 2% Custom B-27 and 1% N-2 supplement (Life Technologies), mouse noggin 10 ng / mL, recombinant human Dkk-1 10 ng / mL, recombinant human IGF-1 10 ng / mL, and recombinant human basic FGF 5 ng / mL (Life Technologies)], and plated in a Matrigel-coated 6-well dish (the same dish as above). The culture medium was changed every other day. From day 10 to 17 of culture (including), the PPC differentiation medium was supplemented with taurine (Sigma-Agentur) and triiodothyronine (T3, Sigma-Agentur) 20 mM and 40 ng / mL, respectively, and feeding was continued according to the same schedule.

[0170] AAGP TM Processing of PPCs 24 hours before cell transplantation, on the day of treatment, the required amount of AAGP TM AAGP prepared by dissolving the powder in a basic cell culture medium (DMEM / F12, 2% B-27 and 1% N-2 supplements, 1% sodium pyruvate, and 1% non-essential amino acids). TM PPCs were treated with 4 mg / mL. All components were purchased from Life Technologies. pH was adjusted by adding 1N NaOH. The pH of the medium was confirmed using Whatman pH test strips (range pH 4.5 to 10). After pH adjustment, AAGP TMThe culture medium was sterilized by filter sterilization (0.22 μm Millipore syringe filter).

[0171] Remove the PPC medium and then add 4 mg / mL AAGP. TM The culture medium was replaced with one containing [specific compound]. The control group of cells was AAGP. TM The same basic medium, without any added substances, was used. The cell culture plates were incubated at 37°C, 5% CO2, and humidity supplied by distilled water in a tray at the bottom of the incubator.

[0172] Cell labeling and transplantation AAGP TM After incubation for 24 hours with or without the specified solution, the PPCs were labeled with Cell Trace Far Red DDAO-SE (Life Technologies, Inc.) according to the manufacturer's specifications. After labeling, the cells were triple-filtered (TrypLE). TM Dissociation was performed using Express Enzyme (Life Technologies). Basic cell culture medium was aspirated and stored. 1 mL triple TM The solution was added to each well, and the cells were incubated for 3 minutes. The collected medium was then returned to the wells to neutralize the trypsin. The cells were harvested, centrifuged at 1500 rpm for 5 minutes, the supernatant was discarded, and the cells were resuspended in sterile Dulbecco's PBS (Life Technologies).

[0173] Cell viability was determined using trypan blue. Before transplantation, labeled cells were divided into 10 cells. 6 The solution was adjusted to a concentration of live cells / mL and then stored on ice.

[0174] 200,000 cells were aseptically injected into the subretinal space in which retinal degeneration had been pre-induced. The handling of the animals and the injection procedure were the same as those described in the "Induction of Retinal Degeneration" section above.

[0175] Tissue processing, immunohistochemistry, and confocal imaging The animals were euthanized 6 months after cell transplantation. After euthanasia, the recipient's eyes were marked with a blue dye for orientation, then excised and fixed in 4% paraformaldehyde at 4°C for 24 hours. These were then cryopreserved, consisting of three 5-minute washes in PBS and incubation in 30% sucrose (in PBS) at 4°C for 24 hours. For neurosensory retinal flat mounts, the retina was bluntly dissected from these eye cups, and the tissue was superfrosted. TM Flatten it on a microscope slide (Fischer), and then attach it to a DAPI Fluoromount-G. TM The PPCs were obtained by mounting them under a coverslip (Southern Biotech). The PPCs were visualized using a fluorescence microscope, and their number was qualitatively evaluated visually.

[0176] After flat-mount preparation, immunostaining for synaptic markers (rib eye and bassoon) and photoreceptor markers (arrestin and blue opsin) was performed according to standard protocols. The tissues were re-fixed with 4% paraformaldehyde at room temperature for 5 minutes, then frozen overnight at 4°C in 30% sucrose (in PBS), and subsequently polyfreezed. TM It was embedded in the culture medium (Polyscience). Immunolabeling was applied using Superfrost. TM The procedure was performed using frozen retinal sections approximately 10 μm thick, placed on microscope slides. The sections were air-dried at room temperature for 1 hour, and then blocked at room temperature for 1 hour with blocking buffer (PBS, 2% normal goat serum, 0.1% Triton X-100). After this, the sections were incubated overnight at 4°C with a suitable primary antibody diluted in the blocking buffer.

[0177] After thoroughly washing with 0.1% Twin 20 in PBS (three times, 10 minutes each), the sections were incubated for 1 hour with a fluorescent secondary antibody diluted in PBS containing 0.1% Twin 20. After washing three times with 0.1% Twin 20 in PBS (as described above), the nuclei were counterstained with DAPI Fluoromount-G. Confocal images were acquired using a Zeiss LSM 510 META confocal laser scanning system.

[0178] The following staining and image processing conditions were used for each marker:

[0179] Arrestincon Primary antibody: Anti-arrestin (catalog number MAB5580; Millipore; dilution 1:500, incubated overnight at 4°C). Secondary antibody: Goat anti-mouse IgG(H+L) cross-adsorbed secondary antibody, Alexa Flure 488 (catalog number A11001; Invitillogen; dilution 1:200; incubation at room temperature for 1 hour). Excitation: 488 nm

[0180] Bassoon dyeing Primary antibody: Anti-Basoon (Catalog No. D63B6; Cell Signaling Technologies; Dilution 1:150, incubated overnight at 4°C). Secondary antibody: Goat anti-rabbit IgG(H+L) cross-adsorbed secondary antibody, Alexa Flure 488 (catalog number A11008; Invitrogen; dilution 1:200; incubation at room temperature for 1 hour). Excitation -488nm

[0181] Rib-eye dyeing Primary antibody: ANTI-CTBP2 clone 16 / CTBP2(RUO) (Catalog number) 612044; BD Transduction Laboratories; Dilution 1:100, 4°C (Incubation overnight). Secondary antibody: Goat anti-mouse IgG(H+L) cross-adsorbed secondary antibody, Alexa Flure 488 (catalog number A11001; Invitrogen; dilution 1:200; incubation at room temperature for 1 hour). Excitation -488nm

[0182] Blue Opsin Staining Primary antibody: Anti-opsin (catalog number AB5407; Millipore; dilution 1:500, incubated overnight at 4°C). Secondary antibody: Goat anti-rabbit IgG(H+L) cross-adsorbed secondary antibody, Alexa Flure 488 (catalog number A11008; Invitrogen; dilution 1:200; incubation at room temperature for 1 hour). Excitation -488nm

[0183] Nuclear staining Reagent: DAPI Fluoromount-G (Catalog No. 0100-20; Southern Biotech; 1 or 2 drops depending on the area). Excitation: 405nm

[0184] Transplanted cells (cell traced before transplantation) TM (Pre-labeled with Far Red DDAO-SE (Life Technologies Inc.)) Excitation - 633nm

[0185] All markers were examined using a fluorescence microscope and qualitatively evaluated by visual inspection. The flowcharts for rabbit and tissue processing are shown in Figure 3.

[0186] Results and Discussion AAGP TM The cell viability of the retinas of rabbits transplanted with treated PPCs and control rabbits (PPCs without AAGP treatment) was analyzed. (PPC and PPC+AAGP) TM Representative images of rabbit retinas from the treatment group are shown in Figure 1. The results obtained show that, after the initial period of 35 days from the start of the study, even without immunosuppression with cyclosporine, 4 mg / mL AAGP TM This study demonstrates that in vitro treatment of PPCs leads to a significant improvement in cell viability 6 months after transplantation of PPCs into the subretinal region of rabbits with retinal degeneration.

[0187] Additional tests were performed to characterize the functional activity and identity of the transplanted PPCs, including fluorescent immunolabeling of arrestin and blue opsin as representative photoreceptor markers, and bassoon and ribeye as example synaptic markers. Immunostaining analysis was completed for selected animals.

[0188] conclusion In conclusion, this study demonstrated that in vitro pretreatment of PPCs with 4 mg / mL AAGP resulted in a significant improvement in cell viability at 6 months after cell transplantation into the subretinal region of rabbits with retinal degeneration. These results suggest that, contrary to expectations, AAGP TM Pretreatment using this method is thought to suppress the immune rejection response of transplanted cells and prevent the onset of immune rejection.

[0189] Example 2 Immunohistochemistry for the Detection of CD4+ T Cells Mouse eyeballs with intact conjunctiva were suspended in an optimal cutting temperature (OCT) compound and flash-frozen in liquid nitrogen. Immunohistochemistry was performed on 6 μm frozen sections, and CD4 (clonal H129.9, 10 μg / mL; BD Biosciences) was used to determine the CD4 concentration. (R) Catalog number 553647), Biotinylated secondary antibody (BD Farmingen) (R) Catalog number 559286), and Nova Red (R) Peroxidase (Vector Laboratories) (R) The number of cells in conjunctival epithelium stained positively with catalog number SK-4800 was detected and counted. Secondary antibody alone and anti-rat isotype (clone A110-2; BD Biosciences) (R) A control group (catalog number 553992) was also examined. Positively stained cells were analyzed using image analysis software (Nikon NIS-Elements) with a 10x objective lens. (R) T cells were counted in GC-rich regions of the conjunctiva using software. T cell detection and quantification were performed only on arms 1, 2, 4, and 5.

[0190] CD4+ T cell infiltration CD4+ T cell infiltration into the conjunctival epithelium is a key clinical indicator of dry eye disease (DED) and is used here as a surrogate to study immune cell infiltration into diseased tissues suffering from retinal degeneration. The presence of CD4+ T cells in the conjunctival epithelium was significantly increased in DED-induced mice without treatment (Figure 5A and B; *, p=0.0130; unpaired student t study). Vehicle administration did not reduce T cell infiltration, but bilateral topical administration of 5% PKX-001 significantly reduced T cell infiltration in DS-induced mice (Figure 5A and B; Table 2; ***, p≦0.0001; unpaired student t study). [Table 2]

[0191] The results of the CD4+ T cell infiltration assay were unexpectedly AAGP TM This indicates that it suppresses the T cell response and has a direct impact on the immune system. This ability to act on immune cells is quite surprising and completely unexpected.

[0192] References 1) Yanai et al., 2015. Efficient generation of photoreceptor progenitor cells from human embryonic stem cells. Methods in Molecular Biology, 1307:357-369. 2) Yanai et al., 2013. Differentiation of human embryonic stem cells using size-controlled embryoid bodies and negative cell selection in the production of photoreceptor progenitor cells. Tissue Engineering, Part C Methods, 19(10):755-764.

[0193] Preferred embodiments have been described above and shown in the accompanying drawings, but it will be apparent to those skilled in the art that modifications can be made without departing from the present disclosure. Such modifications are considered as possible variations that constitute the scope of the present disclosure.

Claims

1. The use of a gem-difluorinated C-glycopeptide compound represented by formula III, or a pharmaceutically acceptable addition salt, hydrate, or solvate of the compound represented by formula III, for the purpose of suppressing or preventing immunorejection of an isolated graft that has come into contact with the compound before implantation into a subject requiring such use, The isolated grafts are photoreceptor progenitor cells, The subject requiring the above-mentioned treatment is receiving immunosuppressant therapy to suppress or prevent immune rejection of the graft, The immunosuppressant therapy is discontinued after a sufficient amount of time has elapsed to transplant the graft into the subject requiring it. Furthermore, the time required to transplant the graft is approximately one to two weeks, or at least two weeks. 【Chemistry 1】

2. The use according to claim 1, wherein the immunosuppressant is sirolimus, tacrolimus, cyclosporine, everolimus, or a combination thereof.

3. The use according to any one of claims 1 to 2, wherein the subject is a human subject.

4. The use according to any one of claims 1 to 3, wherein the isolated graft is isolated from a living donor, a deceased donor, or a combination thereof.

5. The use according to any one of claims 1 to 4, wherein the isolated graft is brought into contact with the compound represented by formula III in a concentration of about 0.01 mg / mL to about 5 mg / mL.

6. The use according to claim 5, wherein the isolated graft is brought into contact with the compound represented by formula III in a concentration of about 1 mg / mL to about 5 mg / mL.

7. The use according to any one of claims 1 to 6, wherein the isolated graft is brought into contact with the compound represented by formula III for about 1 minute to about 1 hour before transplantation.

8. The use according to any one of claims 1 to 6, wherein the isolated graft is brought into contact with the compound represented by formula III for about 12 to about 24 hours prior to transplantation.

9. The use according to any one of claims 1 to 8, wherein the subject has received immunosuppressant therapy for at least about one week prior to the transplantation of the graft.

10. The use according to any one of claims 1 to 9, wherein the isolated graft is washed to remove the compound represented by formula III.

11. The use according to claim 1 for the treatment of retinal degenerative disease.

12. The use according to claim 11, wherein the retinal degenerative disease is age-related macular degeneration (AMD), retinitis pigmentosa (RP), retinal vasculitis, or sarcoidosis.

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