Use of crazakizumab to desensitize HLA-sensitized patients and improve kidney transplantation in those patients.

Abstract

Methods for desensitization of patients requiring organ transplantation are provided. Human leukocyte antigen-sensitized patients awaiting incompatible kidney transplants who are treated with clazakizumab exhibit reduced or absent levels of donor-specific antibodies, indicating improved transplant rates. Clazakizumab and variants are provided for use in various embodiments of the method. In some embodiments, clazakizumab or a variant thereof is administered simultaneously or sequentially with intravenous immune globulin.

Description

[Technical Field]

[0001] Cross-reference of related applications This application includes a claim of priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62 / 757,676 filed on 8 November 2018 and U.S. Provisional Patent Application No. 62 / 855,988 filed on 1 June 2019 (these applications in their entirety are incorporated herein by reference).

[0002] The present invention relates to treatment and procedure methods for desensitization in sensitized patients and for improving organ transplantation. [Background technology]

[0003] All publications within this specification are incorporated by reference to the same extent that individual publications or patent applications are explicitly and individually indicated as being incorporated by reference. The following description contains information that may be useful in understanding the invention. Not all information provided herein constitutes prior art or relates to the currently claimed invention, nor is it acknowledged that any publication explicitly or implicitly referenced is prior art.

[0004] The reasons for accelerating the decline of kidney allografts are multifactorial. Recent data have overturned the long-held view that toxicity of calcineurin inhibitors (CNIs) was the main cause of most chronic allograft failures. Currently, it is recognized that most of the approximately 5,000 annual kidney allograft failures in the United States are due to alloimmune responses. The costs associated with failed kidney allografts represent a significant economic burden on the healthcare system and further reduce the lifespan and quality of life of patients. Patients who return to the transplant waiting list after an allograft failure currently represent the fourth largest category on the new patient waiting list in the United States. This patient population is highly sensitized to human leukocyte antigens (HLAs) and poses a major problem for transplant centers because they are unlikely to receive another transplant without significant desensitization. Currently, there are no FDA-approved drugs in this category. The development of novel therapies to reduce allo-sensitization and improve transplant rates is of great importance. Today, this is one of the most important goals in transplantation medicine.

[0005] HLA molecules are polymorphic. Each HLA molecule expresses polymorphic private epitopes and a number of public determinants that represent epitopes shared by multiple HLA molecules. Immunization following past transplantation, blood transfusion, or pregnancy can trigger the generation of HLA-specific antibodies. An important responsibility of the immunogenetics laboratory is to identify and analyze HLA-specific antibodies present in the sera of patients before and after transplantation. Knowing the specificity of alloantibodies can help predict the likelihood of finding a compatible donor for crossmatch, avoid transplantation from donors with HLA antigens to which the patient is sensitized, help select the optimal crossmatch method, and / or help avoid false-positive crossmatches with donors by excluding clinically irrelevant antibodies.

[0006] Antibodies against HLA antigens strongly influence the mediation of allograft injury and loss and remain a persistent and often formidable obstacle to successful transplantation for thousands of patients on kidney transplant lists worldwide. Preformed or de novo donor-specific antibodies (DSAs) activate complement, induce endothelial cell proliferation, mediate antibody-dependent cell cytotoxicity (ADCC), whereby the recipient becomes highly HLA-sensitized and suffers from a persistent immune attack against the allograft, leading to the progression of interstitial fibrosis, tubular atrophy (IF / TA), as well as allograft dysfunction and loss. Patients who return to dialysis have little hope of receiving another transplant and often have a higher risk of death during dialysis. DSAs are also known to promote atherosclerosis in allografts, thus accelerating renal vascular death.

[0007] To increase the rate of kidney transplantation in sensitized patients, new protocols for HLA desensitization have emerged. This approach requires the application of intravenous immunoglobulin (IVIG), rituximab, and plasma exchange (plasmapheresis, PLEX). There is growing interest in developing new, less expensive, and more convenient immunomodulators to improve antibody reduction at the time of transplantation.

[0008] Existing IVIG-related therapies primarily consist of two desensitization regimens: low-dose intravenous immunoglobulin with plasma exchange (IVIG / PLEX) and high-dose IVIG (HD-IVIG). IVIG / PLEX has been successfully used in ABO-incompatible and positive crossmatch (+CMX) living donor kidney transplants, while HD-IVIG is used to desensitize both living donor + CMS recipients on the waiting list and high-HLA-sensitized postmortem donor (HS-DD) recipients. HD-IVIG (2g / kg) in multi-dose regimens is considered a reasonable approach for desensitization. The B-cell scavenger rituximab is often used in combination with the HD-IVIG and IVIG / PLEX protocols. Rituximab in the IVIG / rituximab protocol has been shown to modify alloreactive B cells to prevent DSA rebound.

[0009] The main problem with existing desensitization regimens lies in interpreting CMX and DSA results when IVIG and rituximab are present in the test serum. IVIG administered at doses of 2 mg / kg (up to 140 g) is likely to interfere with LUMINEX testing for DSA, leading to false-positive results. This can theoretically be avoided by waiting at least one month after IVIG administration before performing LUMINEX single antigen bead (LSA) testing, given that IVIG has a half-life of 30-40 days. Rituximab does not interfere with the LSA testing platform but can produce "false-positive" CDC+ and flow cytometry crossmatch (FCMX) positive B-cell crossmatches that may be misinterpreted as being due to DSA. Pronase treatment of B cells prior to FCMX and CDC testing generally reduces the effect of rituximab, but this cannot always be relied upon.

[0010] Alloantibodies are a major obstacle to access and success in life-saving organ transplantation. Despite advances in desensitization, designing efficient and effective means to eliminate pathogenic anti-HLA antibodies remains a significant medical challenge. Existing desensitization protocols have many notable shortcomings. For example, current treatments carry a risk of acute rejection because they cannot substantially eliminate DSA before transplantation. There is also a risk of rebound DSA formation after transplantation, accompanied by acute and chronic damage to the allogeneic graft. Some current protocols (particularly those using complement inhibitors to prevent chronic antibody-mediated rejection (cABMR)) still fail to achieve desirable outcomes. Therefore, there is an unmet medical need to improve the ability to reduce or eliminate existing HLA antibodies to a level that would allow patients to receive life-saving organ transplantation. [Overview of the project] [Problems that the invention aims to solve]

[0011] Therefore, an object of the present invention is to provide a composition for use in combination with existing standard desensitization treatments, for improving such desensitization treatments, or for use as an alternative to such desensitization treatments, in order to improve the solid organ transplantation rate of HLA-sensitized subjects. [Means for solving the problem]

[0012] The embodiments and aspects described below are illustrative and not limiting in scope, and are described and illustrated together with compositions and methods that are intended to illustrate.

[0013] A method is provided for reducing donor-specific antibodies in human leukocyte antigen (HLA)-sensitized human subjects and / or for desensitization in such subjects. The method comprises clazakizumab; the IL-6 binding fragment of clazakizumab; or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively.H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The method comprises administering an effective amount of a polypeptide having a polypeptide to a subject who requires or has undergone a solid organ transplant. In various embodiments, the subject is provided to be human.

[0014] In one embodiment, crazakizumab, IL-6 conjugated fragment of crazakizumab, or polypeptide disclosed herein is administered before transplantation. In another embodiment, crazakizumab, IL-6 conjugated fragment of crazakizumab, or polypeptide is administered after transplantation. In yet another embodiment, it is provided that crazakizumab, IL-6 conjugated fragment of crazakizumab, or polypeptide is administered both before and after transplantation.

[0015] In some embodiments of the methods of the present disclosure, in addition to the administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide as disclosed herein, a standard therapeutic procedure is provided, including administration of intravenous immunoglobulin (IVIG), rituximab, plasma exchange, or a combination thereof. In one embodiment, this standard therapeutic procedure is administered before the administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide. In another embodiment, this standard therapeutic procedure is administered simultaneously with or after the administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide.

[0016] In one embodiment, the method is provided for desensitizing an HLA-sensitized human patient awaiting kidney transplantation, and the method comprises administering an effective amount of crazakizumab, IL-6 conjugated fragment of crazakizumab, or polypeptide as disclosed herein. In another embodiment, the method for desensitizing an HLA-sensitized human patient awaiting kidney transplantation is provided, comprising administering an effective amount of (1) crazakizumab, IL-6 conjugated fragment of crazakizumab, or polypeptide as disclosed herein, (2) standard therapeutic treatment, e.g., IVIG, plasma exchange, rituximab, or a combination thereof, and optionally (3) an anti-infective agent.

[0017] In other embodiments, one or more of the methods are provided to desensitize HLA-sensitized human patients for the transplantation of other solid organs, including the heart, liver, lungs, pancreas, or intestines.

[0018] In one embodiment, crazakizumab, its IL-6 conjugated fragment, or polypeptide is administered subcutaneously in 1, 2, 3, 4, 5, or 6 doses before transplantation and in 4, 5, 6, 7, 8, 9, 10, 11, or 12 doses after transplantation, at an average dose of approximately 1–5, 5–10, 10–20, or 20–30 mg / dose, in subjects with a decreased amount of donor-specific antibodies after treatment compared to before treatment. In various embodiments, it is provided that crazakizumab, its antigen-conjugated fragment, or polypeptide disclosed herein be administered at intervals of approximately once a month after transplantation. In one embodiment, it is provided that a single dose of crazakizumab, IVIG, and plasma exchange is administered to a subject before transplantation, followed by 6 or 12 doses of crazakizumab after transplantation. In various embodiments, the methods of the Disclosure are provided to administer crazakizumab, its antigen-binding fragment, or polypeptide as disclosed herein to a human subject who is HLA-sensitized and in need of or has undergone a kidney transplant, wherein the subject's creatinine level is reduced after the treatment compared to before the treatment, the presence of donor-specific antibodies is absent or undetectable, and / or the subject has no detectable symptoms or evidence of antibody-mediated rejection (e.g., no deterioration of allograft function as measured by serum creatinine and putative glomerular filtration rate; no detectable evidence of capillary inflammation, inflammation, or complement (C4d) deposition). Further embodiments of the embodiments are provided to provide that the subject's creatinine level is reduced and maintained at a reduced level for one, two, three, four, five months, or longer, concurrently with or after the administration of crazakizumab, its antigen-binding fragment, or polypeptide as disclosed herein.

[0019] Pharmaceutical compositions are also provided for use in administration to HLA-sensitized subjects to desensitize them and increase transplant rates. These pharmaceutical compositions comprise crazakizumab, an IL-6 bound fragment of crazakizumab, or a polypeptide as disclosed herein, and pharmaceutically acceptable excipients (e.g., amino acids, sorbitol, and diluents).

[0020] In various embodiments, the use of crazakizumab is provided in patients who are HLA-sensitized and awaiting incompatible kidney transplants, wherein DSA, complement-dependent cell-mediated cytotoxicity (CDC), and / or antibody-dependent cell-mediated cytotoxicity (ADCC) are reduced or eliminated (e.g., DSA is reduced or eliminated in serum). In some embodiments, patients treated with crazakizumab are suitable for transplantation and have a low likelihood of antibody response.

[0021] In some embodiments, one or more of the methods of the present disclosure are provided to further include selecting a mammalian (e.g., human) patient who is HLA-sensitized and awaiting a kidney transplant from an unsuitable post-mortem (DD) or living donor (LD).

[0022] Other features and advantages of the present invention will become apparent from the following detailed description, along with the accompanying drawings illustrating various features of embodiments of the present invention.

[0023] Exemplary embodiments are shown in the reference drawings. The embodiments and drawings disclosed herein are intended to be considered illustrative rather than restrictive. [Brief explanation of the drawing]

[0024] [Figure 1] The DSA profile for the study involving "ClazaDES01" is shown. This subject is a 50-year-old African American woman with a history of end-stage renal failure (ESRD) secondary to focal segmental glomerulosclerosis (FSGS) as confirmed by biopsy, who has been on dialysis since November 2008 (i.e., an approximately 10-year waiting period for B+ blood type), and her calculated panel-reactive antibody (cPRA) was 58%. The patient's sensitization events included four pregnancies and blood transfusions. [Figure 2]The DSA profile of subject "ClazaDES05" before and after transplantation is shown (median fluorescence intensity, MFI). Subject "ClazaDES05" is a 36-year-old woman with a history of ESRD secondary to IgA nephropathy, who has been on dialysis since June 2008 (i.e., an approximately 10-year waiting period for A+ blood type), and has a cPRA of 100%. The patient's sensitization events included previous transplants and blood transfusions. Subject "ClazaDES05" received a kidney transplant from a post-mortem donor after four doses of crazakizumab. The patient had two types of DSA (Class I and Class II) before and after transplantation. In the case of Class I, the DSA intensity decreased from MFI > 12,500 at transplantation to MFI = 0 at 10 days post-transplantation, and in the case of Class II, the DSA intensity decreased from MFI > 17,500 at transplantation to MFI > 3250 at 10 days post-transplantation. Following the trial protocol, patients received monthly crazakizumab for six months after transplantation. [Figure 3A] This shows the overall amount of C-reactive protein in the crazakizumab desensitization study. Overall, C-reactive protein (CRP) decreased from baseline to nearly zero over 2 months. The number of subjects included in the analysis at each time point is shown in parentheses. [Figure 3B] This shows the levels of individual C-reactive proteins in the crazakizumab desensitization study from baseline to 7 months. [Figure 4] The sum of MFI over time from before plasma exchange (PLEX) (pre-PLEX) to the 5th dose of crazakizumab is shown (N=9). Typically, MFI tends to recover approximately 1–3 months after completion of PLEX / IVIG. Here, with monthly crazakizumab injections, the sum of MFI continued to decrease over time compared to pre-PLEX. To date, three patients have undergone transplantation. Patients ClazaDES01 and ClazaDES03 underwent transplantation after the first dose of crazakizumab. Patient ClazaDES05 underwent transplantation after the 4th dose of crazakizumab. [Figure 5]This outlines exemplary methods for desensitizing HLA-sensitized patients before kidney transplantation. Patients receive up to six doses of crazakizumab, while simultaneously monitoring anti-HLA antibody (DSA levels), Treg cells, and plasmablasts at selected time points during the study. For example, DSA levels are collected at all time points, including day 0, except day 7. C-reactive protein (CRP) and quantitative immunoglobulin (QIC) levels are collected at all time points, including baseline (-15 days), except day 0. In addition, the following are collected at baseline (-15 days) and day 180: CD4+ / CD25+ / Fox P3+ / CD127 low cell count (Tregs); Th17+ cell count; and CD19+ / CD38+ / CD27+ / IL-6+ (plasmablasts). For subjects receiving transplantation before day 180, specialized examinations are performed pre-transplant. For patients receiving transplants 180 days prior to transplantation, specialized tests are performed on day 0 before transplantation. For maintenance, the standard regimen includes tacrolimus, mycophenolate mofetil, and steroids. [Figure 6]This outlines exemplary methods for post-transplant prophylaxis and / or treatment to reduce donor-specific antibodies. In the post-transplant conditioning configuration illustrated in Figure 5, IVIG and crazakizumab are administered post-transplant (in Figure 6, transplant day is day 0). DSA levels are monitored on days 0, 90, and 180, and on day 270 in those receiving a second dose. CRP and QIG levels are collected on days 0, 30, 60, 90, 120, 150, and 180, and on days 240 and 300 in those receiving a second dose. At 180 days post-transplant (approximately 6 months), the following levels are collected: CD4+ / CD25+ / Fox P3+ / CD127 low cell count (Tregs); Th17+ cell count; and CD19+ / CD38+ / CD27+ / IL-6+ (plasmablasts). Viral PCT testing (for cytomegalovirus, Epstein-Barr virus, polyomavirus, BK virus, JC virus, and parvovirus B19) will be performed on days 30, 90, and 180, and on days 270 and 330 if the patient has received a second dose. For maintenance, the standard regimen will include tacrolimus, mycophenolate mofetil, and steroids. "Additional plans" will include continuing monthly crazakizumab for six additional doses (approximately on days 180, 210, 240, 270, 300, and 330) if the patient shows stability or improvement in a) 6M protocol biopsy Banff 2015 read; b) glomerular filtration rate (GFR); and c) DSA. [Figure 7] This shows the timeline of treatment for patient "ClazaDES03" in the example study, and the patient's creatinine level (mg / dL) from before treatment to after treatment. [Figure 8] This image shows a kidney transplant biopsy of patient "ClazaDES03" approximately two months after transplantation, including tubular injury, arteriosclerosis, and very localized tubulitis. [Figure 9]This image shows a kidney transplant biopsy of patient "ClazaDES03" approximately 6 months post-transplant (including acute tubular necrosis, a rare isometric vacuole of the same size, and mild tubulointerstitial inflammation). [Figure 10] Figures 10A and 10B show the flow panel reactivity antibody test (flow-PRA) class I / class II results before desensitization and after transplantation (post-transplantation). [Figure 11] This report shows the HLA class I and class II antibody levels and total levels of various markers in DES03 patients treated with crazakizumab, both before desensitization and at the final follow-up (F / U) approximately 6–12 months post-transplant. DSA was not detected at the time of transplantation or post-transplant. [Figure 12-1] Figures 12A-12C show the HLA class I and class II antibody levels and total levels of various markers in DES05 subjects treated with crazakizumab, both before desensitization and after transplantation. [Figure 12-2] Continuation of Figure 12-1. [Figure 13-1] Figures 13A-13C show the HLA class I and class II antibody levels and total levels of various markers in DES07 subjects treated with crazakizumab, both before desensitization and after transplantation. [Figure 13-2] Continuation of Figure 13-1. [Figure 14] This shows the HLA class I & class II antibody levels and total doses for various markers in the target DES02 (non-transplanted) before desensitization and after crazagizumab. [Figure 15] This shows the HLA class I & class II antibody levels and total doses for various markers in DES09 (non-transplanted) before desensitization and after crazagizumab. [Figure 16] This shows HLA class I and class II antibody levels before and after crazakizumab for all study patients (N=10). [Figure 17-1] Figures 17A-C show HLA class I antibodies (Figure 17A) and class II antibodies (Figure 17B) before and after crazakizumab in transplant patients (N=8) (compared together in Figure 17C). [Figure 17-2] Continuation of Figure 17-1. [Figure 18] DSA for individual patients regarding Class I and Class II before desensitization, at the time of transplantation, and after transplantation is shown (N=8). [Figure 19] The mean DSA MFI for Class I and Class II before desensitization, at the time of transplantation, and after transplantation is shown. [Modes for carrying out the invention]

[0025] All references cited herein are incorporated by reference in their entirety as if they were listed in their entirety. Unless otherwise defined, technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this invention pertains. Singleton et al., Dictionary of Microbiology and Molecular Biology 3 rd ed., Revised, J. Wiley & Sons (New York, NY 2006); March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 7 th ed., J. Wiley & Sons (New York, NY 2013); and Sambrook and Russel, Molecular Cloning: A Laboratory Manual 4 th The ed., Cold Spring Harbor Laboratory Press (Cold Spring Harbor, NY 2012) provides a general guide to many of the terms used in this application. For reference on methods of antibody preparation, see D. Lane, Antibodies: A Laboratory Manual 2. nded. (Cold Spring Harbor Press, Cold Spring Harbor NY, 2013); Kohler and Milstein, (1976) Eur. J. Immunol. 6:511; Queen et al. U.S. Patent No. 5,585,089; and Riechmann et al., Nature 332:323 (1988); U.S. Patent No. 4,946,778; Bird, Science 242:423-42 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); Ward et al., Nature 334:544-54 (1989); Tomlinson I. and Holliger P. (2000) Methods Enzymol, 326, 461-479; Holliger See P.(2005)Nat.Biotechnol.Sep;23(9):1126-36).

[0026] Those skilled in the art will recognize many methods and substances similar to or equivalent to those described herein that may be used in the practice of the present invention. In fact, the present invention is by no means limited to the methods and substances described herein. For the purposes of the present invention, the following terms are defined below.

[0027] The term "transplant rate" generally refers to the number of patients who receive a transplant for every 100 patients on a waiting list in a given year. In some embodiments, this transplant rate is a measure of how frequently patients on a program's waiting list receive a transplant. To facilitate comparison of numbers, in some embodiments, this rate is expressed as "per 100 patients / year," meaning that the rate is normalized to represent the number of patients who would receive a transplant if there were 100 patients on the list over a year. For example, a transplant rate of 5 transplants per 100 patients / year means that 5 transplants are performed for every 100 patients on the list over a year. Because this is a normalized rate, the number may contain decimals, such as 5.1 transplants per 100 patients / year. This means that it is expected that slightly more than 5 patients per 100 patients on the list will receive a transplant over a year.

[0028] A positive crossmatch (+CMX) indicates the presence of donor-specific alloantibodies (DSAs) in the potential recipient's serum and is often associated with a graft loss rate exceeding 80%.

[0029] In the examples, “HLA-sensitized (HS) patients” refer to patients on the United Network for Organ Sharing (UNOS) waiting list awaiting a kidney transplant who have a calculated panel-reactive antibody (cPRA) or a probability of being a crossmatched incompatible donor of 50% or higher, who also have a demonstrable DSA using LUMINEX bead technology in various embodiments, and who have a history of sensitization events (e.g., previous transplant, blood transfusion, and / or pregnancy). The presence of HLA-specific antibodies can be determined by testing the patient's serum against cells from a panel of HLA-type donors or against soluble HLA antigens attached to a solid support. Generally, HLA-sensitized patients refer to patients with cPRA of 10%, 20%, 30%, 40%, or 50% or higher.

[0030] "Subject" means human or animal. Typically, this animal is a vertebrate such as a primate, rodent, domesticated animal, or game animal. Examples of primates include chimpanzees, crab-eating macaques, spider monkeys, and macaques, such as rhesus macaques. Examples of rodents include mice, rats, woodchucks, ferrets, rabbits, and hamsters. Examples of domesticated and game animals include cattle; horses, pigs, deer, bison, buffalo, feline species (e.g., domestic cats), and canid species (e.g., dogs, foxes, wolves). The terms "patient," "individual," and "subject" are used interchangeably herein. In some embodiments, the subject is an animal. This animal may be, but is not limited to, a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse, or a cattle. In addition, domesticated animals and / or pets may be treated using the methods described herein.

[0031] The terms “treat,” “treatment,” “treating,” or “relieve” refer to therapeutic treatments aimed at reversing, reducing, relieving, inhibiting, delaying, or halting the progression or severity of a condition associated with a disease or disorder. The term “treat” includes reducing or mitigating at least one adverse effect or symptom of a condition, disease, or disorder (e.g., weight loss or muscle loss resulting from cancer cachexia). A treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, a treatment is “effective” if the progression of the disease is slowed or stopped. That is, “treatment” includes not only improvement of symptoms or markers but also halting at least a delay in progression or worsening of symptoms that would be expected if no treatment were performed. Beneficial or desirable clinical outcomes include, but are not limited to, relief of one or more symptoms, reduction of the severity of the disease, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and detectable or undetectable (partial or overall) remission. The term “treatment” of the disease also includes bringing about relief of the symptoms or side effects of the disease (including palliative treatment).

[0032] The term "antibody" refers to intact immunoglobulins or monoclonal or polyclonal antigen-binding fragments that have the Fc (fragment crystallizable) region or the FcRn-binding fragment of the Fc region, also referred to herein as the "Fc fragment" or "Fc domain". The antigen-binding fragments may be produced by recombinant DNA techniques or by enzymatic or chemical cleavage of intact antibodies. As antigen-binding fragments, inter alia, Fab, Fab’, F(ab’)2, Fv, dAb, and complementarity-determining region (CDR) fragments, single-chain antibodies (scFv), single-domain antibodies, chimeric antibodies, bispecific antibodies, and polypeptides that contain at least a portion of an immunoglobulin sufficient to confer specific binding of an antigen to the polypeptide are mentioned. The Fc domain includes a portion of two heavy chains that contribute to two or three classes of antibodies. The Fc domain may be produced by recombinant DNA techniques or by enzymatic cleavage (e.g., papain cleavage) or chemical cleavage of intact antibodies. The antibody can be a chimeric antibody, a humanized antibody, or a human antibody. The antibody can be an IgG1 antibody, an IgG2 antibody, an IgG3 antibody, or an IgG4 antibody. In some embodiments, the antibodies herein have an Fc region that is modified to alter at least one of effector function, half-life, proteolysis, or glycosylation.

[0033] The term "antibody fragment" refers to a protein fragment that contains only a portion of an intact antibody and generally contains the antigen-binding site of the intact antibody and thus retains the ability to bind to an antigen. Examples of antibody fragments included in this definition are as follows: (i) a Fab fragment having a V L domain, a C L domain, a V H domain, and a CH1 domain; (ii) a Fab’ fragment that is a Fab fragment having one or more cysteine residues at the C-terminus of the CH1 domain; (iii) an Fd fragment having a V H domain and a CH1 domain; (iv) a V HFd' fragment having a domain and a CH1 domain, with one or more cysteine ​​residues at the C-terminus of the CH1 domain; (v) V of a single arm of the antibody L Domain and V H Fv fragment having a domain; (vi)V H (vii) a dAb fragment consisting of domains; (viii) an isolated CDR region; (viii) a bivalent F(ab')2 fragment containing two Fab' fragments linked by disulfide crosslinks in the hinge region; (ix) a single-chain antibody molecule (e.g., single-chain Fv, scFv); (x) a light chain variable domain (V) in the same polypeptide chain. L ) connected to the heavy chain variable domain (V H (xi) A "bispecific antibody" having two antigen-binding sites, including (xi) a pair of tandem Fd segments (V) that form a pair of antigen-binding domains together with a complementary light chain polypeptide. H -CH1-V H A "linear antibody" containing -CH1). The antibody or antibody fragment may be scFv, camelbody, nanobody, IgNAR (single-chain antibody derived from shark), and Fab fragment, Fab' fragment, or F(ab')2 fragment.

[0034] "Selectively binding" or "specifically binding" refers to the K of the antibody or antibody fragment described herein. D 10 -5 M (10000 nM) or less (for example, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 This refers to the ability to bind to a target (e.g., a molecule present on the cell surface) at M (or lower). Specific binding may be affected, for example, by the affinity and binding activity of the polypeptide agent, as well as the concentration of the polypeptide agent. Those skilled in the art can determine the appropriate conditions under which the polypeptide agents described herein selectively bind to a target using any appropriate method, such as titration of the polypeptide agent in a suitable cell binding assay.

[0035] An “ineffective” treatment means that when the treatment is applied to the subject, there is less than a 5% improvement in symptoms. Where specifically stated in the claims, an ineffective treatment may mean less than 1%, 2%, 3%, 4%, 6%, 7%, 8%, 9%, or 10% improvement in symptoms.

[0036] An adverse event is any undesirable and unintended sign, symptom, or illness temporarily associated with the use of the drug under study (IMP) or any intervention imposed by the protocol, regardless of the cause. An adverse event may be any undesirable and unintended sign (abnormal laboratory findings), symptom, or illness temporarily associated with the use of the drug, regardless of whether it is considered to be related to the drug. Surgical intervention is not an adverse event, but a therapeutic measure for a condition requiring surgery. However, a condition requiring surgery is an adverse event if it occurs or is detected during the study in this embodiment. Planned surgical interventions and conditions leading to such interventions are not adverse events if the condition was known before the commencement of the procedure in the study. In the latter case, the condition should be reported as part of the medical history.

[0037] Pre-existing medical conditions are those present at the start of this study. Any worsening of a pre-existing condition during this study will be considered an adverse event. A pre-existing condition should be recorded as an adverse event if its frequency, intensity, or characteristics worsen during the study period.

[0038] An abnormal laboratory finding that meets any one of the following criteria should be considered an adverse event: The test results are related to the accompanying symptoms; The test results indicate that additional diagnostic tests or medical / surgical intervention are necessary; The test results may lead to changes in the administration of the test treatment (e.g., changes in dosage, interruption, or permanent discontinuation), or concomitant medication treatment (e.g., addition, interruption, or discontinuation), or any other changes in the associated medication or treatment; The test results lead to any of the outcomes included in the definition of a serious adverse event (Note: This would be reported as a serious adverse event); The test results are considered an adverse event by the principal investigator.

[0039] Test results that are outside the normal range and do not meet any of the above criteria should not be reported as adverse events. Repeated abnormal tests in the absence of the above conditions do not constitute an adverse event. Any abnormal test result determined to be an error does not need to be reported as an adverse event.

[0040] A "serious adverse event" (SAE) is any adverse medical occurrence at any dose as follows: Fatal (leading to death); Life-threatening: When an adverse event occurred, the patient was at immediate risk of death from that adverse event. This does not include events that occurred in a more severe form or that, if continued, could have resulted in death. Hospitalization is required, or an extension of existing hospitalization is necessary; To result in a persistent or significant disability / loss of ability; This is a congenital anomaly / congenital defect (in the case of a child of a patient exposed to this test procedure); A significant medical event that is not fatal, not life-threatening, or does not require hospitalization may be considered a SAE if, based on appropriate medical judgment, the event may endanger the subject and require medical or surgical intervention to prevent one of the outcomes listed in this definition. Examples of such events include emergency room or home intensive care for non-hospitalizing allergic bronchospasm, blood disorders, or seizures, or for the onset of drug dependence or abuse.

[0041] Adverse events leading to hospitalization are considered serious. Any adverse event leading to initial hospitalization or transfer to an acute / intensive care unit within a hospital is considered serious.

[0042] Hospitalization or extension of hospitalization without a triggering clinical adverse event is not a serious adverse event in itself. Examples of hospitalizations that are not considered serious adverse events include: (1) hospitalization for treatment of an existing condition not related to the onset of a new adverse event or exacerbation of an existing condition; (2) hospitalization for social or administrative reasons; (3) hospitalization for reasons not related to a triggering clinical adverse event; and (4) pre-planned procedures or surgeries, which should not be documented in the baseline documentation.

[0043] All adverse events require severity assessment by researchers. Severity is assessed using the following criteria: Mild: You notice some discomfort, but it does not interfere with your normal daily activities. Moderate: Discomfort sufficient to reduce or affect daily activities; and Severe: Unable to work or perform daily activities.

[0044] To clarify the difference between the terms "critical" and "severe," which are not synonyms, it should be noted that the term "severe" is often used to describe the intensity (severity) of a specific event, such as a mild, moderate, or severe myocardial infarction. However, the event itself may have relatively minor medical significance, such as a severe headache. This is not identical to "critical," which is based on criteria of patient / event outcome or behavior that typically pose a threat to a patient's life or function. Severity (not severity) serves as a guideline for defining reporting obligations by regulatory authorities.

[0045] The assessment of causality is to determine whether there is a reasonable possibility that the treatment in this study caused the adverse event or contributed to it. “Not related” explains that the temporal connection with the administration of the treatment in this study is lost or unbelievable, or that there is evidence of another cause. “Low likelihood related” explains that the causal relationship from the temporal connection with the administration of the treatment in this study is unbelievable, and that another drug, chemical, or underlying condition provides a more plausible explanation. “Possibly related” explains a reasonable temporal sequence with the administration of the treatment in this study, but the event could also be explained by a comorbidity with another drug or chemical. Information regarding drug withdrawal may be insufficient or unclear. “Strongly related” explains a plausible temporal connection with the administration of the treatment in this study, and the event could not be explained by a comorbidity or another drug or chemical. Responses to drug discontinuation (drug administration interruption) should be clinically reasonable. This event must be confirmed pharmacologically or phenomenologically using sufficient rechallenge procedures as necessary.

[0046] Interleukin-6 is a key mediator of inflammation, as well as the development, maturation, and activation of T cells, B cells, and plasma cells. Excessive IL-6 production is associated with many human diseases characterized by excessive and uncontrolled antibody production and autoimmunity.

[0047] Methods of the present disclosure for desensitizing HLA-sensitized subjects, for reducing the amount of donor-specific antibodies, and / or improving organ transplant rates or transplant survival rates include administering to a subject an effective amount of an antibody or its antigen-binding fragment that shares at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence homology (identity) to crazakizumab or the complementarity-determining region (CDR) of crazakizumab. Some embodiments provide that one or more of these methods further include administering an effective amount of IVIG or plasma exchange.

[0048] Clazakizumab is a glycosylated humanized monoclonal antibody (from a rabbit parent antibody) that targets interleukin-6. Peptide sequence and structural information for crazakizumab is available from IMGT / mAb-db record #414. Peptide sequence analysis by BLAST reveals a complete agreement with the peptide claimed in U.S. Patent No. 8,062,864, which is incorporated herein by reference in its entirety. Further descriptions of crazakizumab and its variants are provided in U.S. Patent No. 7,935,340, which is incorporated herein by reference in its entirety, and this antibody or antibody fragment is suitable for the methods disclosed herein for reducing or removing donor-specific antibodies in subjects who need or are undergoing allograft transplantation. For example, this antibody is used in Sequence ID No. 1 (V H CDR1), Sequence ID 2 or 3 (V H CDR2), and Sequence ID 4 (V H V contains the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide, respectively, which are included in CDR3. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. LIt contains a polypeptide. This anti-human IL-6 antibody contains a variable heavy chain contained in SEQ ID NO: 8, 9, or 10 and a variable light chain contained in SEQ ID NO: 11 or 12. Asn-Tyr-Tyr-Val-Thr (Sequence ID 1) Ile-Ile-Tyr-Gly-Ser-Asp-Glu-Thr-Ala-Tyr-Ala-Thr-Trp-Ala-Ile-Gly (SEQ ID NO: 2) Ile-Ile-Tyr-Gly-Ser-Asp-Glu-Thr-Ala-Tyr-Ala-Thr-Ser-Ala-Ile-Gly (SEQ ID NO: 3) Asp-Asp-Ser-Ser-Asp-Trp-Asp-Ala-Lys-Phe-Asn-Leu (Sequence ID 4) Gln-Ala-Ser-Gln-Ser-Ile-Asn-Asn-Glu-Leu-Ser (Sequence ID 5) Arg-Ala-Ser-Thr-Leu-Ala-Ser (Sequence ID 6) Gln-Gln-Gly-Tyr-Ser-Leu-Arg-Asn-Ile-Asp-Asn-Ala (Sequence ID 7).

[0049] The variable heavy chain sequence is described below: Sequence ID 8-METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNY YVTWVRQAPGKGLEWIGIIYGSDETAYATWAIGRFTISKTSTTVDLKMTS LTAADTATYFCARDDSSDWDAKFNLWGQGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVK.

[0050] The substituted variable heavy chain sequence is described below: SEQ ID NO: 9-EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATWAIGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNL.

[0051] Another substituted variable heavy chain sequence is listed below: Sequence ID 10-EVQLVESGGGLVQPGGSLRLSCAASGFSLSNYYVTWVRQAPGKGLEWVGIIYGSDETAYATSAIGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDSSDWDAKFNL.

[0052] The variable light chain sequence is described below: SEQ ID NO: 11-MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQS INNELSWYQQKPGQRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDL ECADAATYYCQQGYSLRNIDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNN.

[0053] The substituted variable light chain sequence is described below: SEQ ID NO: 12-IQMTQSPSSLSASVGDRVTITCQASQSINNELSWYQQKPGKAPKLLIYRASTLASGVPSRFSGSGSGTDFTLTISSLQPDDFATYYCQQGYSLRNIDNA.

[0054] Clazakizumab is a genetically engineered humanized immunoglobulin G1 (IgG1) antibody that binds to human IL-6 with an affinity of approximately 4 pM. Using multiple assays on signaling and cellular function in response to IL-6 alone (to measure classical signaling) and in combination with IL-6 and sIL-6R (to measure transsignaling), crazakizumab has been demonstrated to be a potent and complete antagonist of IL-6-induced signaling, as measured by phosphorylation of signaling factors and activator 3 (STAT3), as well as cellular functions such as cell proliferation, differentiation, activation, immunoglobulin B-cell production, and hepatocyte production of acute-phase proteins (C-reactive protein [CRP] and fibrinogen). In addition, crazakizumab has been shown to be a competitive antagonist of IL-6-induced cell proliferation.

[0055] Clazakizumab may address destructive alloantibody reactions to allogeneic grafts and exhibits broad immunomodulatory effects that may be useful as a desensitizer to improve kidney transplant rates in highly HLA-sensitized patients. Clazakizumab is widely evaluated in patients with rheumatoid arthritis but has not yet been approved by the FDA for all conditions. Since the introduction of IL-6 / IL-6R blockers, inhibition of the IL-6 / IL-6R pathway may be significantly beneficial in systemic lupus erythematosus (SLE) and other vasculitic disorders, with reduced antibody-producing cells reported in treated patients. Currently, there is no information on clazakizumab for highly sensitized patients awaiting incompatible (HLAi) transplants or for the treatment of antibody-mediated rejection.

[0056] To date, despite studies of crazakizumab in healthy subjects, as well as in subjects with rheumatoid arthritis (RA), psoriatic arthritis (PsA), Crohn's disease, graft-versus-host disease (GVHD), and tumors, no studies of crazakizumab have been conducted in highly sensitized patients who have undergone kidney transplantation.

[0057] Various embodiments are one or more methods for reducing donor-specific antibodies in HLA-sensitized subjects, characterized in that the calculated panel-reactive antibody (cPRA) or the proportion of likely crossmatch-incompatible donors is at least 10%, 20%, 30%, 40%, 50%, 60%, or 70%, and the subjects are pre-kidney transplant, post-kidney transplant, or both before and after kidney transplant, and include crazakizumab; its antigen-binding fragment; or V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. LThe present invention provides a method comprising administering an effective amount of a polypeptide having a polypeptide. Various embodiments of this method provide administering an effective amount of an anti-human IL-6 antibody or antibody fragment comprising the variable heavy chain of SEQ ID NO: 8, 9, or 10 and the variable light chain of SEQ ID NO: 11 or 12 to an HLA-sensitized subject before or after allograft transplantation to reduce or eliminate donor-specific antibodies in the subject after allograft transplantation. Some embodiments of this method further include selecting a subject in which the calculated panel-reactive antibody (cPRA) or the percentage of likely crossmatch-unsuitable donors is at least 10%, 20%, 30%, 40%, 50%, 60%, or 70%. Some embodiments of this method further include performing a kidney transplant in the subject. Further embodiments of this method are characterized by a decrease in donor-specific antibodies after kidney transplantation due to the administration of crazakizumab or its antigen-binding fragment; or by an undetectable amount of donor-specific antibodies beginning about one, two, three, four, five, or six months after kidney transplantation due to the administration of crazakizumab or its antigen-binding fragment.

[0058] Various embodiments include one or more methods for reducing donor-specific antibodies in HLA-sensitized subjects, comprising (1) an effective amount of IVIG, and (2) crazakizumab; the IL-6 binding fragment of crazakizumab; or the polypeptide of CDR1 contained in SEQ ID NO: 1, the polypeptide of CDR2 contained in SEQ ID NO: 2 or 3, and the polypeptide of CDR3 contained in SEQ ID NO: 4. H A polypeptide containing the polypeptide CDR1 contained in SEQ ID NO: 5, the polypeptide CDR2 contained in SEQ ID NO: 6, and the polypeptide CDR3 contained in SEQ ID NO: 7. L The present invention provides a method comprising administering an effective amount of a polypeptide having a polypeptide. In one embodiment, IVIG is administered to crazakizumab; the IL-6 binding fragment of crazakizumab; or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. HPolypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The polypeptide is administered before or concurrently with the polypeptide. In some embodiments, the HLA-sensitized subject is to receive a solid organ transplant, in one embodiment this solid organ is derived from a crossmatched donor (i.e., the HLA-sensitized subject contains pre-transplant antibodies against HLA derived from the donor's organ), and in another embodiment this solid organ is not derived from a positive crossmatched donor. In other embodiments this one or more methods include (1) an effective dose of IVIG, and (2) crazakizumab; the IL-6 binding fragment of crazakizumab; or V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L In addition to administering an effective amount of polypeptide containing polypeptide, the procedure further includes performing solid organ transplantation.

[0059] Some embodiments of a method for reducing donor-specific antibodies in HLA-sensitized subjects include: (1) an effective amount of a combination of IVIG and crazakizumab; (2) an effective amount of a combination of IVIG and crazakizumab IL-6 binding fragments; or (3) IVIG and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L This involves administering an effective amount of a combination of polypeptides containing a polypeptide. In one embodiment, IVIG is administered to crazakizumab; the IL-6 binding fragment of crazakizumab; or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. HPolypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The polypeptide is administered before or simultaneously with the polypeptide having the polypeptide. In another embodiment, IVIG is further administered immediately before, during, or immediately after solid organ transplantation in the subject.

[0060] Further embodiments include a method for reducing donor-specific antibodies in an HLA-sensitized subject, comprising: (1) an effective dose of a combination of IVIG, plasma exchange, and crazakizumab; (2) an effective dose of a combination of IVIG, plasma exchange, and IL-6 binding fragments of crazakizumab; or (3) IVIG, plasma exchange, and a combination of CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The present invention provides a method comprising administering an effective amount of a combination of polypeptides having a polypeptide. In one embodiment, IVIG and plasma exchange are administered using crazakizumab; IL-6 binding fragment of crazakizumab; or CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide, which are contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L Administer before or simultaneously with polypeptides containing polypeptides.

[0061] In various embodiments, crazakizumab-induced desensitization (1) led to transplantation in 8 out of 10 patients in the example study, (2) led to a significant decrease in HLA specificity, although cPRA did not change significantly, and (3) led to a decrease in DNA at and after transplantation with continued administration of effective doses of crazakizumab.

[0062] In further embodiments, anti-IL-6 treatment (e.g., crazakizumab administration) significantly reduces MFI levels of class II / class II HLA antibodies, thereby increasing the transplant rate in HLA-sensitized patients or increasing the likelihood or rate of transplant survival in individual HLA-sensitive patients. Anti-IL-6 mediates this by reducing the number of plasma cells that produce IL-6 (anti-HLA). After treatment, anti-IL-6 therapy reduces or eliminates DSA levels and prevents DSA generation in de novo. Further embodiments also provide that patients receiving crazakizumab and solid organ transplants do not develop antibody-mediated rejection of the transplant.

[0063] Various embodiments of the present disclosure provide that the method includes identifying HLA-sensitized patients requiring solid organ transplantation, administering PLEX and IVIG and a monthly dose of crazakizumab, performing solid organ transplantation (e.g., kidney transplantation), administering induction therapy such as alemtuzumab and / or THYMOGLOBULIN (anti-thymocyte globulin), and administering immunosuppressive therapy such as tacrolimus, CELLCEPT (mycophenolate mofetil), and tapering prednisone. In one embodiment, the transplantation rate for HLA-sensitized patients after desensitization with crazakizumab and PLEX / IVIG is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%. In another embodiment, the time from completion of pre-transplant desensitization with crazakizumab and PLEX / IVIG to transplantation in HLA-sensitized subjects is 0 days to 1 week, 1 week to 1 month, 1 month to 3 months, 3 months to 6 months, 6 months to 1 year, 1 year to 2 years, or longer. In a further embodiment, subjects who have undergone desensitization treatment with an effective dose of crazakizumab (or possibly in combination with PLEX / IVIG) and have undergone an allogeneic kidney transplant, and who are already HLA-sensitized, have no signs or symptoms of antibody-mediated rejection of the transplant in at least 95%, 90%, 85%, 80%, 75%, or 70% of cases. A further embodiment of this method includes the fact that the subjects do not have rheumatoid arthritis (RA), psoriatic arthritis (PsA), Crohn's disease, graft-versus-host disease (GVHD), cancer, or a combination thereof. A further aspect of this method further includes selecting subjects who do not have or have never had rheumatoid arthritis (RA), psoriatic arthritis (PsA), Crohn's disease, graft-versus-host disease (GVHD), or cancer, who are HLA-sensitive, and who require or have received a solid organ (e.g., kidney) transplant, for a method of reducing and / or removing donor-specific antibodies.

[0064] Various embodiments provide that one or more of the methods of the Disclosure further include performing one or more assays on a subject before and / or after allograft transplantation for the presence or absence of infection associated with cytomegalovirus, Epstein-Barr virus, polyomavirus, BK virus, JC virus, parvovirus B19, or a combination thereof. In other embodiments, one or more of the methods of the Disclosure are characterized in that, before and / or after allograft transplantation, the subject does not have a detectable amount of infection associated with cytomegalovirus, Epstein-Barr virus, polyomavirus, BK virus, JC virus, parvovirus B19, or a combination thereof. Further embodiments provide that a subject in one or more of the post-transplant clazakizumab methods of the Disclosure is examined for the absence of chronic antibody-mediated rejection of a solid organ transplant or for the absence of evidence of chronic antibody-mediated rejection.

[0065] Various embodiments of a method for reducing or removing donor-specific antibodies in an HLA-sensitized subject requiring or undergoing allograft transplantation, and / or desensitizing such HLA-sensitized subject, include administering, in one or more doses over time, an effective amount of a polypeptide comprising crazakizumab, the IL-6 binding fragment of crazakizumab, a polypeptide comprising the variable heavy chain of SEQ ID NO: 8, 9, or 10 and the variable light chain of SEQ ID NO: 12 or 12, or a polypeptide comprising the variable heavy chain having CDR1 of SEQ ID NO: 1, CDR2 of SEQ ID NO: 2 or 3, and CDR3 of SEQ ID NO: 4, and the variable light chain having CDR1 of SEQ ID NO: 5, CDR2 of SEQ ID NO: 6, and CDR3 of SEQ ID NO: 7, (1) (1) The subject has or has had pre-formed donor-specific antibodies (DSA) prior to allograft transplantation, and / or has developed or has developed DSA after allograft transplantation; (2) The subject has a calculated panel-reactive antibody level of 50% or higher; (3) The subject has high donor-specific antibody intensity, as determined, for example by a single-antigen LUMINEX bead assay, expressed as greater than 9,000, 10,000, 11,000, or 12,000 for class I or class II, or higher mean fluorescence intensity (MFI); or (4) The subject has experienced one or more pregnancies, blood transfusions, and / or previous transplants. In one embodiment, the subject has one of the characteristics mentioned. In another embodiment, the subject has two of the characteristics mentioned. In yet another embodiment, the subject has three of the characteristics mentioned. In yet another embodiment, the subject has four of the characteristics mentioned.

[0066] Additional embodiments of the methods disclosed herein include one or more steps of immunomonitoring before and / or after allograft transplantation. In various embodiments, a method for reducing or eliminating donor-specific antibodies in a subject who had pre-formed DSA, cPRA of 50% or more, or high-intensity DSA prior to allograft transplantation, wherein the subject subsequently underwent allograft transplantation (e.g., the allograft is HLA-incompatible with the subject), and the method comprises (i) administering an effective amount of crazakizumab, IL-6 conjugated fragment of crazakizumab, or the polypeptide disclosed above in one or more doses; and (ii) administering individually over one or more periods (e.g., 1 month, 2 months, 3 months, 4 months, 5 months, Over periods of 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 15 months, 18 months, 24 months, or longer, after one or more doses of crazakizumab, crazakizumab IL-6 conjugate fragment, or this polypeptide, (a) immunomonitoring of this subject, such as assaying blood samples of this subject to quantify Treg, Tfh, Th17, B cells, IL-6, CRP, plasma cells, plasmablast IgG levels, or combinations thereof; (b) post-transplant biopsy evaluation; (c) measurement of glomerular filtration rate; and / or (d) measurement of the amount of DSA in this subject;Furthermore, if applicable, (iii)(a) immunomonitoring indicates improved immunoresponsiveness (e.g., lower levels of CRP, Treg, Tfh, Th17, B cells, IL-6, plasma cells, or plasmablast IgG compared to previous immunomonitoring or baseline measurements at or before allograft transplantation, or equivalent levels of CRP, Treg, Tfh, Th17, B cells, IL-6, plasma cells, or plasmablast IgG compared to healthy or desensitized subjects), (b) biopsy evaluation of transplantation indicates the absence of cell-mediated and antibody-mediated rejection, the absence of or reduced evidence of allograft dysfunction (e.g., determined by C4d staining and transplant glomerulopathy using the Banff 2015 criteria), and / or Banff If improvement is observed in accordance with the 2015 criteria, (c) if the glomerular filtration rate is stable (e.g., at a similar or decreased level compared to the last measurement or pre-transplant), or (d) if the DSA level is stable (e.g., at a similar or lower level compared to the last measurement or pre-transplant), further administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide may be discontinued or limited to no further six months; if immunomonitoring indicates, for example, that the immunoreactivity described above has not improved or that the glomerular filtration rate or DSA level is not stable, administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide may be continued or continued at an adjusted dose; if biopsy evaluation of the transplant indicates the presence of cell-mediated rejection and / or antibody-mediated rejection, this may include treating the rejection with standard therapeutic measures (e.g., IVIG, plasmapheresis, or both). In some cases, steps (ii) and (iii) may be repeated one, two, three, four, five, six, seven, eight, nine, or ten times, or as needed, or until improvement, stabilization, or even healing is observed.

[0067] In some embodiments, if evidence of antibody-mediated rejection is observed, the subject is guided to treatment for antibody-mediated rejection. In some embodiments, after steps (i) and (ii), no further doses of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide are administered for a period of time such as one week, two weeks, three weeks, four weeks, two months, and three months ("rest period"), and after the "rest period," immunoreactivity is monitored or graft biopsy is evaluated, and depending on the results of characterization in step (iii), a person skilled in the art would discontinue or continue administration of crazakizumab, crazakizumab IL-6 conjugate fragment, or polypeptide to further reduce or eliminate DSA in the subject.

[0068] Dosage In one embodiment, a method for reducing donor-specific antibodies and desensitizing HLA in a subject (e.g., a human subject) includes administering an effective dose of crazakizumab or an antigen-binding fragment of crazakizumab pre-transplant (e.g., approximately 25 mg / dose subcutaneously every four weeks over up to six doses). In another embodiment, a method for reducing donor-specific antibodies and desensitizing HL in a subject (e.g., a human subject) includes administering plasma exchange (or plasmapheresis) five, six, or seven times pre-transplant, followed by administering an effective dose of IVIG (e.g., at a maximum of approximately 2 g / kg of subject in 140 g doses) and an effective dose of crazakizumab (e.g., approximately 25 mg subcutaneously every four weeks over up to six doses). In a further embodiment, the method includes transplanting an allogeneic graft into the subject. In a further embodiment, allogeneic graft transplantation occurs between the last dose of crazakizumab and approximately 270 days after IVIG administration. This is shown in Figure 5. In one embodiment, the effective dose of crazakizumab to reduce DSA levels in HLA-sensitized subjects is approximately 12.5 mg / dose over 4, 5, 6, or more doses. In another embodiment, the effective dose of crazakizumab to reduce DSA levels in HLA-sensitized subjects is approximately 25 mg / dose over 4, 5, 6, or more doses. In yet another embodiment, the effective dose of crazakizumab to reduce DSA levels in HLA-sensitized subjects is not 25 mg / dose in a monthly dose over 4, 5, or 6 doses.

[0069] In another embodiment, a method for reducing donor-specific antibodies and desensitizing HLA in a subject (e.g., a human subject) includes administering an effective dose of crazakizumab or its antigen-binding fragment post-transplant (e.g., subcutaneously at approximately 25 mg every four weeks, starting at approximately 5-7 days post-transplant, over up to six doses). In a further embodiment, the method includes administering an additional effective dose of crazakizumab (e.g., over six further doses, up to 330 days post-transplant). This is illustrated in Figure 6. In one embodiment, an effective dose of crazakizumab to reduce DSA levels after solid organ transplantation in an HLA-sensitized subject is approximately 12.5 mg / dose over one, two, three, four, five, or more doses. In another embodiment, an effective dose of crazakizumab to reduce DSA levels after solid organ transplantation in an HLA-sensitized subject is approximately 25 mg / dose over one, two, three, four, five, or more doses. In one embodiment, the effective dose of crazakizumab to reduce DSA levels after solid organ transplantation in HLA-sensitized subjects is not 25 mg / dose administered every four weeks.

[0070] In another embodiment, a method for reducing donor-specific antibodies and desensitizing HL in a subject (e.g., a human subject) includes administering an effective dose of crazakizumab or an antigen-binding fragment of crazakizumab before transplantation (e.g., subcutaneously at approximately 25 mg / dose every 4 weeks over up to 6 doses), and administering an effective dose of crazakizumab or an antigen-binding fragment after transplantation (e.g., subcutaneously at approximately 25 mg every 4 weeks, starting approximately 5-7 days post-transplant, over up to 6 doses).

[0071] Some embodiments of these methods provide assaying biopsies from patients and confirming stable levels of glomerular filtration rate (GFR) over a long period (e.g., fluctuations of less than 10%, 20%, or 30% over two, three, or four consecutive biopsies) and low levels of DSA (e.g., less than 10%, 20%, or 30%) compared to before desensitization treatment or to previous biopsies performed after transplantation. In some embodiments, if the GFR level is unstable or the DSA level is high, the method further includes repeated administrations of effective doses of IVIG and crazakizumab until the GFR level stabilizes and the DSA level decreases.

[0072] Another embodiment provides a method for reducing donor-specific antibodies and HLA sensitization in a highly HLA-sensitized subject (e.g., a human subject) comprising: performing plasma exchange (or plasmapheresis) before transplantation; administering an effective dose of IVIG before, during, or after transplantation; and administering an effective dose of crazakizumab to the subject before, after, or both, wherein the subject has a stable level of glomerular filtration rate (GFR) over a long period (e.g., less than 10%, 20%, or 30% variation over two, three, or four consecutive biopsies) and a low level of DSA (e.g., less than 10%, 20%, or 30%) compared to before the desensitization treatment.

[0073] Following administration of crazakizumab as a 1-hour intravenous infusion, the pharmacokinetics of crazakizumab were linear, as indicated by consistent clearances at these dose levels, across the 30 mg to 640 mg dose range in healthy subjects and across the 80 mg to 320 mg dose range in subjects with rheumatoid arthritis (RA). The T-half of crazakizumab at all doses was very similar in healthy male subjects and subjects with RA, and was consistent with what is expected for humanized IgG1 antibodies. Across the doses tested, the mean T-half of crazakizumab ranged from 19.5 to 31.0 days in healthy male subjects and from 26.4 to 30.9 days in subjects with RA. The T-half of crazakizumab after SC administration in healthy male subjects was similar to that after IV administration. In a phase 1 trial comparing IV and SC administration in healthy men, the mean T-half of crazakizumab was 30.7 days after a single IV dose and 31.1–33.6 days after SC administration. The bioavailability of crazakizumab after SC administration was 60% of that of the IV formulation. As expected, Cmax was lower and Tmax was longer with SC administration compared to IV administration.

[0074] Population PK analysis of clinical trial data in RA, PsA, and healthy subjects shows that body weight influences crazakizumab PK, with both clearance and central distribution volume increasing with increasing body weight. Therefore, heavier subjects have lower drug exposure compared to lighter subjects.

[0075] The effective dose of crazakizumab for each participant may be considered or limited based on safety assessments. Safety assessments include interviews, adverse event recording, physical examination, blood pressure, and laboratory measurements. Participants will generally be evaluated for adverse events (all grades), serious adverse events, and adverse events requiring interruption or discontinuation of the study drug at each trial visit during their participation in this study.

[0076] In some embodiments, a V containing crazakizumab, an IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively, is suitable for administration by the method of the present disclosure. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The effective dose of the polypeptide containing the polypeptide is approximately 10-50 μg / dose, 50-100 μg / dose, 100-150 μg / dose, 15 doses, or more, over the total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 doses, or, if necessary, to continue reducing the amount of DSA in the subject. 0~200μg / dose, 100~200μg / dose, 200~300μg / dose, 300~400μg / dose, 400~500μg / dose, 500~600μg / dose, 600~70 0μg / dose, 700~800μg / dose, 800~900μg / dose, 900~1000μg / dose, 1000~1100μg / dose, 1100~1200μg / dose, 1200~1 300μg / dose, 1300~1400μg / dose, 1400~1500μg / dose, 1500~1600μg / dose, 1600~1700μg / dose, 1700~1800μg / dose Amount, 1800~1900μg / dose, 1900~2000μg / dose, 2000~2100μg / dose, 2100~2200μg / dose, 2200~2300μg / dose, 2300~ The dose may be in the range of 2400 μg / dose, 2400-2500 μg / dose, 2500-2600 μg / dose, 2600-2700 μg / dose, 2700-2800 μg / dose, 2800-2900 μg / dose, or 2900-3000 μg / dose, and may be administered once daily, once weekly, once every two weeks, once a month, or once every two months, or a combination thereof.

[0077] In some embodiments, per unit weight of the target in the above method, V contains crazakizumab, an IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively, suitable for administration by the method of this disclosure. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L Effective amounts of polypeptides containing polypeptides include 10-100 μg, 100-200 μg, 200-300 μg, 300-400 μg, 400-500 μg, 500-600 μg, 600-700 μg, 700-800 μg, 800-900 μg, 1-5 mg, 5-10 mg, 10-20 mg, 20-30 mg, 30-40 mg, 40-50 mg, 50-60 mg, 60-70 mg, 70-80 mg, 80-90 mg, 90-100 mg, 100-200 mg, 200-300 mg, 300-400 mg, 400 mg-500 mg, 500 mg-1 g, or 1 g-10 g. The unit weight of the subject may be per kg of body weight or per subject. In one embodiment, the effective dose of crazakizumab for reducing and desensitizing DSA levels in already HLA-sensitized human subjects requiring or having undergone allogeneic kidney transplantation is approximately 25 mg / month. In one embodiment, the effective dose of crazakizumab for reducing and desensitizing DSA levels in already HLA-sensitized human subjects requiring or having undergone allogeneic kidney transplantation is not approximately 25 mg / month.

[0078] In further embodiments, a V comprising crazakizumab, an IL-6 binding fragment of crazakizumab, or the CD1R polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively, suitable for administration by the method of the present disclosure. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. LThe effective amount of the polypeptide having the polypeptide may be in the range of 0.01-0.05 mg / kg, 0.05-0.1 mg / kg, 0.1-1 mg / kg, 1-5 mg / kg, 5-10 mg / kg, 10-50 mg / kg, or 50-100 mg / kg. In additional embodiments, the effective amount of crazakizumab, the antigen-binding fragment of crazakizumab, or the polypeptide of the present disclosure may be about 1-2 mg / kg, 2-3 mg / kg, 3-4 mg / kg, 4-5 mg / kg, 5-6 mg / kg, 6-7 mg / kg, 7-8 mg / kg, 8-9 mg / kg, 9-10 mg / kg, 10-11 mg / kg, 11-12 mg / kg, 12-13 mg / kg, 13-15 mg, 15-20 mg / kg, or 20-25 mg / kg. In additional embodiments, the effective dose of crazakizumab, crazakizumab antigen-binding fragments, or the polypeptide of the Disclosure may be one or more of the following: approximately 100–125 mg, 125–150 mg, 150–175 mg, 160–170 mg, 175–200 mg, 155–165 mg, 160–165 mg, 165–170 mg, 155–170 mg, or any combination thereof, and this effective dose may be administered in 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 doses, some of which may be administered pre-transplant and others post-transplant.

[0079] In various embodiments, a V comprising crazakizumab, an IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively, is suitable for administration by the method of the present disclosure. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. LA polypeptide having a polypeptide is administered at least once, 1 to 7 times per week, 1 to 7 times per month, or 1 to 12 times per year, or once or more times as needed, in one or more of the doses described herein, over a period of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 14 months, 16 months, 18 months, approximately 24 months, approximately 30 months, approximately 36 months, or a combination thereof, in one or more doses from among those described herein.

[0080] Pharmaceutical composition In various embodiments, the present invention provides a pharmaceutical composition comprising (1) crazakizumab, an IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L (1) A polypeptide comprising (2) a polypeptide and (3) a pharmaceutically acceptable excipient.

[0081] The pharmaceutical compositions according to the present invention may contain any pharmaceutically acceptable excipients. “pharmaceutically acceptable excipients” generally means excipients that are safe, non-toxic, and useful in the preparation of a desirable pharmaceutical composition, and include excipients that are acceptable not only for human pharmaceutical use but also for veterinary use. Such excipients may be solid, liquid, semi-solid, or, in the case of aerosol compositions, gaseous. Examples of excipients include, but are not limited to, amino acids, starches, sugars, microcrystalline cellulose, diluents, granulators, lubricants, binders, disintegrants, wetting agents, emulsifiers, colorants, release agents, coating agents, sweeteners, flavoring agents, fragrances, preservatives, antioxidants, plasticizers, gelling agents, thickeners, hardeners, setting agents, suspending agents, surfactants, humectants, carriers, stabilizers, and combinations thereof.

[0082] In one embodiment, the method of the present disclosure comprises L-histidine, L-histidine monohydrochloride, sorbitol, polysorbate-80, and water for injection, and V containing crazakizumab, IL-6 binding fragment of crazakizumab, or CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L The method includes administering a pharmaceutical composition comprising a polypeptide having a polypeptide.

[0083] In various embodiments, the pharmaceutical composition according to the present invention may be formulated for delivery by any route of administration. In one embodiment, the pharmaceutical composition is administered intravenously or subcutaneously to a subject. "Route of administration" can refer to any route of administration known in the art (e.g., aerosol, nasal, oral, mucosal, dermal, parenteral, or enteral). "Parareal" refers to a route of administration generally associated with injection (e.g., intraorbital, infusion, intra-arterial, intra-articular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intramedullary, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, mucosal, or transtracheal). By parenteral routes, the composition may be in the form of a solution or suspension for infusion or injection, or in the form of a lyophilized powder. By parenteral routes, the composition may be in the form of a solution or suspension for infusion or injection. By the enteral route, this pharmaceutical composition may be in the form of tablets, gel capsules, sugar-coated tablets, syrups, suspensions, solutions, powders, granules, emulsions, microspheres or nanospheres, or controlled-release lipid vesicles or polymer vesicles. Typically, the composition is administered by injection. These methods of administration are known to those skilled in the art.

[0084] The pharmaceutical compositions according to the present invention may contain any pharmaceutically acceptable carrier. “pharmaceutically acceptable carrier,” as used herein, means a pharmaceutically acceptable substance, composition, or vehicle involved in the transport or delivery of the compound of interest from one tissue, organ, or part of the body to another. For example, a carrier may be a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, or a combination thereof. Each component of the carrier must be “pharmaceutically acceptable” in that it must be compatible with the other components of the formulation. Similarly, it must be suitable for use in contact with any tissue or organ it may come into contact with, meaning it must not carry a risk of toxicity, irritation, allergic reactions, immunogenicity, or any other complications that would outweigh the therapeutic benefits.

[0085] The pharmaceutical compositions according to the present invention may also be encapsulated, tableted, or prepared as emulsions. The compositions may be enhanced or stabilized, facilitated, or result in sustained or controlled release (or increased half-life) of the compositions by adding pharmaceutically acceptable solid or liquid carriers. Examples of liquid carriers include syrup, peanut oil, olive oil, glycerin, saline, alcohol, and water. Examples of solid carriers include starch, lactose, calcium sulfate, dihydrate, clay, magnesium stearate or stearic acid, talc, pectin, acacia, agar, or gelatin. Examples of emulsion carriers include liposomes or controlled-release polymer nanoparticles known in the art. Methods for preparing liposome delivery systems are discussed in Gabizon et al., Cancer Research (1982) 42:4734; Cafiso, Biochem Biophys Acta (1981) 649:129; and Szoka, Ann Rev Biophys Eng (1980) 9:467. Other drug delivery systems are known in this field and are described, for example, in Poznansky et al., DRUG DELIVERY SYSTEMS (RLJuliano, ed., Oxford, NY 1980), pp.253-315; MLPoznansky, Pharm Revs (1984) 36:277. As carriers, sustained-release substances such as glyceryl monostearate or glyceryl distearate, either alone or together with wax, may also be used.

[0086] This pharmaceutical preparation is manufactured according to conventional pharmaceutical techniques (for example, in the case of tablets, crushing, mixing, granulation, and compression if necessary; or in the case of hard gelatin capsules, crushing, mixing, and filling). When a liquid carrier is used, the preparation is in the form of a syrup, elixir, emulsion, or aqueous or non-aqueous suspension. Such liquid formulations may be administered directly by PO or filled into soft gelatin capsules.

[0087] The pharmaceutical composition according to the present invention can be delivered in a therapeutically effective amount. The precise therapeutically effective amount is the amount of composition that yields the most effective result in terms of the efficacy of the treatment in a given subject. This amount varies depending on various factors, including, but not limited to, the properties of the therapeutic compound (e.g., activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological state of the subject (e.g., age, sex, type and stage of disease, overall physical condition, responsiveness to a given dose, and type of drug), the properties of the pharmaceutically acceptable carrier in the formulation, and the route of administration. Those skilled in the art in the clinical and pharmacological fields can determine the therapeutically effective amount by routine experiments, for example, by monitoring the subject's response to the administration of the compound and adjusting the dose accordingly. For further guidance, see Remington: The Science and Practice of Pharmacy (Gennaro ed. 20th edition, Williams & Wilkins PA, USA) (2000).

[0088] Before administering to the patient, the formulation agent (formulant) contains crazakizumab, the IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L It can be added to polypeptides having polypeptides. Liquid formulations may be preferred. For example, this formulation agent may contain oil, polymer, vitamin, carbohydrate, amino acid, salt, buffer, albumin, surfactant, filler, or a combination thereof.

[0089] Examples of carbohydrate preparations include sugars or sugar alcohols (e.g., monosaccharides, disaccharides, or polysaccharides), or water-soluble glucans. Examples of saccharides or glucans include fructose, dextrose, lactose, glucose, mannose, sorbose, xylose, maltose, sucrose, dextran, pullulan, dextrin, alpha-cyclodextrin and beta-cyclodextrin, soluble starch, hydroxyethyl starch, and carboxymethylcellulose, or mixtures thereof. "Sugar alcohol" is defined as a C4-C8 hydrocarbon having an -OH group, and examples include galactitol, inositol, mannitol, xylitol, sorbitol, glycerol, and arabitol. These sugars or sugar alcohols may be used alone or in combination. There is no limit to the amount used, as long as the sugar or sugar alcohol is soluble in the aqueous preparation. In one embodiment, the concentration of sugar or sugar alcohol is 1.0 w / v% to 7.0 w / v%, more preferably 2.0 to 6.0 w / v%.

[0090] Examples of amino acid formulations include the levorotatory (L) forms of carnitine, arginine, and betaine, but other amino acids may also be added.

[0091] In some embodiments, the polymer used as a formulation agent may be polyvinylpyrrolidone (PVP) with an average molecular weight of 2,000 to 3,000, or polyethylene glycol (PEG) with an average molecular weight of 3,000 to 5,000.

[0092] To minimize the pH change of the solution before or after lyophilization, it is also preferable to use a buffer in this composition. Most physiological buffers (e.g., citrate buffer, phosphate buffer, succinate buffer, and glutamate buffer, or mixtures thereof) can be used. In some embodiments, the concentration is 0.01 to 0.3 molar. Surfactants that can be added to this formulation are shown in European Patent Nos. 270,799 and 268,110.

[0093] After preparing a liquid pharmaceutical composition, it can be freeze-dried to prevent degradation and maintain sterility. Methods for freeze-drying liquid compositions are known to those skilled in the art. Immediately before use, the composition can be restored with a sterile diluent that may contain additional components (e.g., Ringer's solution, distilled water, or sterile saline). Once restored, the composition is administered to the subject using methods known to those skilled in the art.

[0094] Antiinfective agents Various embodiments provide that the desensitization method further comprises administering one or more antiinfective agents, preferably post-transplant, as a preventive or therapeutic measure against bacterial, viral, or fungal infections.

[0095] Examples of antiinfective agents suitable for use in the manner of this disclosure include: antibiotics, e.g., aminoglycosides (e.g., amikacin, gentamicin, kanamycin, neomycin, netylmycin, streptomycin, tobramycin, paromomycin), ansamycin (e.g., geldanamycin, harbimycin), carbasephalosporins (e.g., loracarbef), carbapenems (e.g., ertapenem, doripenem, imipenem, cilastatin, meropenem), and ce Pharosporines (e.g., 1st generation: cefadroxil, cefazolin, cefalotin or cefalothin, cephalexin; 2nd generation: cefaclor, cephamandol, cefoxitin, cefprodil, cefuroxime; 3rd generation: cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone; 4th generation: cefepime)Fifth generation: ceftoviprole), glycopeptides (e.g., teicoplanin, vancomycin), macrolides (e.g., azithromycin, clarithromycin, dilithromycin, erythromycin, roxithromycin, troleandmycin, telithromycin, spectinomycin), monobactams (e.g., aztreonam), penicillins (e.g., amoxicillin, ampicillin, azurocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin) Nishilin, piperacillin, ticarcillin), polypeptide antibiotics (e.g., bacitracin, colistin, polymyxin b), quinolones (e.g., ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, ofloxacin, trovafloxacin), rifamycin (e.g., rifampicin or rifampin, rifabutin, rifapentin, rifaximin), sulfonamides (e.g., mafenide, prontosil, sulfacetamide, sulfamethizone) Sulfanilamide, sulfasalazine, sulfisoxazole, trimethoprim, trimethoprim-sulfamethoxazole (cotrimoxazole, "tmp-smx"), and tetracyclines (e.g., demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline), as well as arsphenamine, chloramphenicol, clindamycin, lincomycin, ethambutol, fosfomycin, fusidic acid, furazolidone, isoniazid, linezolid, metronidazole, mupirocin, Nitrofurantoin, platensimycin, pyrazinamide, quinupristin / dalfopristin combination, and tinidazole. In some embodiments, a method for reducing donor-specific antibodies in an HLA-sensitized subject before and / or after allograft transplantation comprises administering to the subject an effective dose of crazakizumab or an IL-6 conjugated antibody fragment of crazakizumab, and administering to the subject an effective dose of ganciclovir, valganciclovir, fluconazole, trimethoprim, sulfamethoxazole, or a combination thereof.

[0096] kit In various embodiments, the present invention provides a kit for desensitizing organ transplant recipients. The kit comprises an assembly of materials or components (e.g., crazakizumab, IL-6 binding fragment of crazakizumab; or V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively). H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L A polypeptide having polypeptides; instructions or manual for administration for desensitization before and after organ transplantation; one or more vessels as containers; and, if applicable, one or more diluents.

[0097] The exact properties of the components comprising the kit of the present invention depend on its intended purpose. In one embodiment, the kit is configured specifically for human use. In further embodiments, the kit is configured for veterinary use and, without limitation, for treating subjects such as livestock, domesticated animals, and laboratory animals.

[0098] This kit may include instructions for use. These instructions typically include tangible representations describing the techniques used in the use of the kit's components to achieve a desired outcome (e.g., to treat or inhibit the cancerous cachexia of interest). In some cases, the kit may also include other useful components, such as measuring instruments, diluents, buffers, pharmaceutically acceptable carriers, syringes, or other useful tools readily recognizable to those skilled in the art.

[0099] The materials or components for assembling this kit may be stored and provided to the practitioner in any convenient and appropriate manner that maintains their operability and practicality. For example, components may be in the form of thawed, dehydrated, or freeze-dried, and may be provided at room temperature, refrigerated, or frozen temperatures. Components are typically housed in appropriate packaging materials. As used herein, the term “packaging materials” refers to one or more physical structures used to house the contents of the kit, such as the compositions and similars of the present invention. Packaging materials are constructed by well-known methods and are preferably configured to provide a sterile and uncontaminated environment. As used herein, the term “packaging” refers to an appropriate solid matrix or material (e.g., glass, plastic, paper, foil, and similar) capable of holding individual kit components. For example, packaging may include crazakizumab, the IL-6 binding fragment of crazakizumab, or the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H Polypeptides and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L A bottle may be used to contain an appropriate amount of the composition of the present invention, which comprises a polypeptide having a polypeptide. The packaging material generally has an external label indicating the contents and / or purpose of the kit and / or its components. [Examples]

[0100] The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the invention. Where specific materials are mentioned, these materials are for illustrative purposes only and are not intended to limit the invention. Those skilled in the art may develop equivalent means or reactants without demonstrating inventive ability and without departing from the scope of the invention.

[0101] Study: A Phase I / II trial to evaluate the safety and tolerability of crazakizumab to improve transplant rates by eliminating donor-specific HLA antibodies in hyper-HLA-sensitized (HS) patients awaiting kidney transplantation. This trial is an open-label design to evaluate the safety and efficacy of crazakizumab in clearing DSA and inducing Treg subsets in HS patients awaiting HLA-mismatched (HLAi) kidney transplantation. An overview of this protocol is shown in Figures 5 and 6. Safety determinations aim to assess all adverse events associated with crazakizumab administration and the risk of infectious complications associated with crazakizumab treatment for desensitization in HS patients awaiting renal HLAi transplantation. Limited efficacy determinations include evaluating the reduction in DSA levels that enables increased transplant rates and the subsequent prevention of ABMR (eGFR, SCr) after crazakizumab treatment.

[0102] The investigational drug, crazakizumab, contains a genetically engineered humanized anti-IL-6 monoclona antibody as its active ingredient. When manufactured at Ajinomoto Althea (San Diego, CA), crazakizumab has a strength of 25 mg / mL. Crazakizumab contains excipients including L-histidine, L-histidine monohydrochloride, sorbitol, polysorbate-80, and sterile water for injection. It is contained in a 25 mg / mL single-dose vial for undiluted injection. The crazakizumab vial should be stored protected from light at -20°C (-4°F) or below. Prepared syringes can be stored in a refrigerator at 2-8°C (36-46°F) ≤ -20°C (-4°F) for up to 24 hours, and up to 4 hours of this 24-hour period can be stored at room temperature at 15-25°C (59-77°F). Prepared syringes should be protected from light. Before administration, crazakizumab should be brought to room temperature by storing it without refrigeration for 30-60 minutes before use.

[0103] procedure This was a Phase I / II clinical trial of crazakizumab in highly HLA-sensitized patients awaiting kidney transplantation. The trial was intended to run for three years. HLA antibodies were detected using a solid-phase assay system currently in use at the Cedars-Sinai Medical Center HLA Laboratory.

[0104] Typically, patients participating in this trial received PLEX (5-7 doses) + IVIG initially, followed by subcutaneous (SC) administration of crazakizumab 25 mg one week after IVIG. If no safety / tolerance / efficacy issues were observed after the initial dose, the patient received five additional injections every four weeks (Q4W). If the patient underwent HLAi transplantation, crazakizumab was continued for six doses (starting on post-transplant day 5) at 25 mg SC Q4W for six months post-transplant. Protocol biopsies were performed at six months post-transplant, and the allograft was evaluated for evidence of ABMR, including C4d staining and TG, using the Banff 2015 criteria. If improvement was observed after the sixth dose of crazakizumab, the patient would continue with another six doses for six months. Patients who presented with evidence of persistent allograft dysfunction may undergo a non-protocol biopsy for this reason. Patients who received 12 doses of crazakizumab after transplantation would undergo a 12M protocol biopsy. If a patient did not show improvement after 6 doses of crazakizumab, they would not receive further treatment, and the patient would return on day 365 for a final trial visit.

[0105] Specifically, a single dose of IVIG was administered at the time of transplantation (day 0) (IVIG #1). A second dose of IVIG was administered on day 1. The treatment period then began. Clazakizumab was administered six times at days 5 ± 2d, 30 ± 7d, 60 ± 7d, 90 ± 14d, 120 ± 14d, and 150 ± 14d, respectively.

[0106] Figure 7 shows the trial timeline for patient ClazaDes03 and his creatinine levels over time. Patient ClazaDES03 is a 32-year-old male with a history of end-stage renal failure secondary to an unclear etiology. He is the post-2012 living donor unrelated kidney transplant recipient, which failed in 2016. In patient ClazaDES03's case, the transplant was performed after the first dose of crazakizumab (25 mg subcutaneously) and before the second dose of crazakizumab. This patient received PLEX (5-7 doses) on day -15; IVIG at 2 g / kg on day 0; post-transplant #1 crazakizumab (25 mg SQ) on day 7 (April 5, 2018); a post-mortem donor kidney transplant was performed on day 21 (April 29, 2018); post-transplant #1-#6 crazakizumab (25 mg SQ) was administered at approximately monthly intervals; alemtuzumab (CAMPATH 1H) was introduced, and maintenance was maintained with tacrolimus, mycophenolate mofetil (MMF), and prednisone. Approximately two months after transplantation, on June 28, 2018, a biopsy of the allogeneic kidney graft was examined (Figure 8), showing sub-best creatinine levels, and monthly administration of beratacept was started on August 14, 2018. A 6M biopsy was performed on October 23, 2018 (Figure 9). Second round (#7-#12) of crazakizumab administration began on October 31, 2018 (#7 post-transplant crazakizumab). In this patient, no donor-specific antibodies were present after transplantation.

[0107] Anti-HLA antibodies may occur naturally or arise from past pregnancies, blood transfusions, or previous transplants. Patients treated with six doses of crazakizumab for desensitization were given blood samples for HLA antibodies, and other monitoring of the blood samples and immunological testing were performed. If a patient underwent an HLAi transplant during this study, they received a standard post-transplant immunosuppression protocol and immunological monitoring, along with subcutaneous administration of crazakizumab 25 mg every four weeks for six doses. Immunological monitoring of blood samples, including Treg, Tfh, Th17, and B cell subsets, as well as IL-6 and CRP monitoring, was performed at the Cedars-Sinai Transplant Immunology Laboratory.

[0108] result Figure 1 shows the DSA profile for the subject "ClazaDES01" in this trial. This subject was a 50-year-old African American woman with a history of end-stage renal failure (ESRD) secondary to focal segmental glomerulosclerosis (FSGS) as confirmed by biopsy, who had been on dialysis since November 2008 (i.e., an approximately 10-year waiting period for B+ blood type), and her calculated panel-reactive antibody (cPRA) was 58%. The patient's sensitization events included four pregnancies and blood transfusions.

[0109] Figure 2 shows the DSA profile of subject "ClazaDES05" before and after transplantation (median fluorescence intensity, MFI). Subject "ClazaDES05" is a 36-year-old woman with a history of ESRD secondary to IgA nephropathy, who has been undergoing dialysis since June 2008 (i.e., an approximately 10-year waiting period for A+ blood type), and has a cPRA of 100%. The patient's sensitization events included previous transplants and blood transfusions. Subject "ClazaDES05" received a kidney transplant from a post-mortem donor after four doses of crazakizumab. The patient had two types of DSA (Class I and Class II) before and after transplantation. In the case of Class I, DSA intensity decreased from MFI > 12,500 at transplantation to MFI = 0 at 10 days post-transplantation, and in the case of Class II, MFI > 17,500 at transplantation to MFI > 3250 at 10 days post-transplantation. Following the trial protocol, patients received monthly crazakizumab for six months after transplantation.

[0110] Figure 3A shows the overall C-reactive protein levels in the clazakizumab desensitization study. Overall, C-reactive protein (CRP) decreased from baseline to nearly zero at 2 months. The number of subjects included in the analysis at each time point is shown in parentheses. Figure 3B shows the individual C-reactive protein levels in the clazakizumab desensitization study from baseline to 7 months.

[0111] Figure 4 shows the total MFI over time from pre-PLEX to the fifth dose of crazakizumab (N=9). Typically, MFI tends to recover approximately 1–3 months after completion of PLEX / IVIG. Here, with monthly crazakizumab injections, the total MFI continued to decrease over time compared to pre-PLEX. To date, three patients have undergone transplantation. Patients ClazaDES01 and ClazaDES03 underwent transplantation after the first dose of crazakizumab. Patient ClazaDES05 underwent transplantation after the fourth dose of crazakizumab.

[0112] Regarding patient "ClazaDES03," Figure 7 shows the patient's creatinine (mg / dL) levels over time, comparing pre-desensitization and post-desensitization and post-kidney transplantation levels. Creatinine levels were maintained at low post-transplant levels after two doses of crazakizumab. Figure 8 shows the 2-month kidney transplant biopsy of patient "ClazaDES03." This biopsy revealed mild acute tubular injury; mild to moderate arteriosclerosis and mild arteriosclerosis (consistent with the donor's disease); no diagnostic evidence of acute rejection (at most the borderline of cell-mediated rejection according to Banff criteria); and mesangial IgA deposition was present, but there were no associated proliferative glomerular changes. Since IgA staining was absent in a biopsy from the patient's previous kidney transplant in 2012, the IgA present in this biopsy was likely donor-related. Figure 9 shows a 6-month kidney transplant biopsy from patient "ClazaDES03". This biopsy showed acute tubular necrosis with rare same-sized vacuoles; mild tubulointerstitial inflammation (at most borderline changes of cell-mediated rejection); arteriosclerosis; and minimal interstitial fibrosis / tubular atrophy. The detection of rare same-sized vacuoles may be related to acute calcineurin inhibitor toxicity from recent IVIG treatment. No diagnostic features of antibody-mediated rejection or polyomaviral nephropathy were present.

[0113] Overall, desensitization treatment using crazakizumab resulted in a reduction in HLA antibodies in at least 40% of patients in this trial, leading to successful transplantation.

[0114] Between March and November 2018, a total of 10 patients were enrolled. Transplantation was approved in 9 patients; 8 patients underwent transplantation during the study period, and the 9th patient received transplantation 2 months after the completion of the study. 4 patients reached the 12-month study period. All administered fluids were well tolerated. No graft loss or patient death was observed, nor were there any serious infections attributable to or requiring discontinuation of crazakizumab. Renal function was stable at 6 months. Demographic and immunological / transplant characteristics are summarized in Tables 1 and 2.

[0115] [Table 1]

[0116] [Table 2]

[0117] All patients except for the ninth underwent a 6-month protocol biopsy. Two patients (25) showed confirmed rejection by biopsy: one with chronic active cell-mediated rejection (Banff grade 1A); and one with chronic active antibody-mediated and cell-mediated rejection (Banff grade 1B). Both patients responded to treatment according to the research center's standard treatment protocol.

[0118] Of the nine patients who underwent transplantation, seven (78%) had DSA before desensitization and at the time of transplantation. Only three patients (33%) had DSA detected at 1 month; two patients (22%) had DSA detected at 3 months; and no patients had DSA detected at 6 months (6M), 9 months (9M), or 12 months (12M) (including patients with DSA detected at 1 month and 3 months). The DSA MFI values ​​(mean ± SD) before desensitization, at transplantation, at 1 month, and at 3 months were as follows: 11060 ± 6990, 7980 ± 6260, 1923 ± 3973, and 1040 ± 2650; and 0 ± 0 at 6M, 9M, and 12M, respectively. The mean DSA MFI was significantly reduced when comparing pre-desensitization with 1M (p=0.0004) and 3M (p=0.0001), and when comparing the time of transplantation with 1M (p=0.007) and 3M (p=0.001).

[0119] Seven SAEs occurred, all of which appeared unrelated to crazakizumab. These SAEs included wound dehiscence requiring resuturing (1 SAE), hematuria and UTIs prior to the first dose of the study drug (1 SAE), as well as bacteremia (1 SAE), thrombosis of the right external iliac artery with graft loss (1 SAE), persistent surgical site pain requiring CT-guided drainage of pre- and post-graft fluid with proliferation of MSSA (1 SAE), chronic active ABMR as evidenced by biopsy as a result of delayed d / t infection and chronic thrombocytopenia following crazakizumab administration (1 SAE), and perirenal effusion requiring CT-guided drainage (1 SAE).

[0120] Primary secondary purpose To determine whether crazakizumab treatment can significantly reduce or eliminate ABMR development and C4d deposition in incompatible allografts transplanted in high-HLA sensitized patients after crazakizumab desensitization. Allograft function was evaluated up to 6–12 months (6–12M) post-transplant, and renal function was determined using serum creatinine (SCr), Modification of Diet in Renal Disease (MDRD) GFR calculation (creatinine clearance (CrCl) estimated using the Schwartz equation for patients under 18 years of age), and DSA levels. Protocol biopsies were performed 6M after crazakizumab treatment. In addition, several immunological measurements of blood samples were evaluated before and after the initiation of crazakizumab treatment. These included: T reg Cells (CD4+, CD25+, FoxP3+CD127 dim ) evaluation T fh Evaluation of cells (CD4+, ICOS+, CXCR5+, IL-21+) Evaluation of circulating plasmablasts (CD19+, CD38+, CD27+, IL-6+) Assessment of CRP and IL-6 levels.

[0121] These secondary endpoints will help understand the biology of the alloimmune response to allografts and determine the capacity and mechanism of the beneficial effects of crazakizumab. Viral PCR was monitored according to standard treatment guidelines.

[0122] Inclusion criteria Age 15–75 years at screening. HS patients (cPRA ≥ 50%) awaiting DD or LD kidney transplant on the UNOS list. History of pregnancy, blood transfusion, and / or kidney transplant. Subjects / parents / guardians must be fully willing to participate in the study requirements. Subjects / parents / guardians must understand and be able to submit informed consent. Pneumococcal vaccination. Negative tuberculin (ppd) placement result or negative Quantiferon TB Gold result. These individuals must also have sufficient waiting time on the UNOS list to allow for frequent applications due to a history of positive crossmatch (DD), or positive flow cytometry (FCMX) mismatch (LD) and negative complement-dependent cell-mediated cytotoxicity (CDC+) crossmatch. Patients proceeding to HLAi transplant after desensitization will have CDC CMX negativity at 1:2 dilution, channel shift of FCMX < 225, and DSA, which is already defined as an acceptable MFI.

[0123] Exclusion criteria Multiple organ transplants (e.g., kidney and pancreas). Intolerance to treatment with crazakizumab or other IL-6 inhibitors. Women who are breastfeeding or pregnant. Women of childbearing age, and their male partners who, during the trial and for five months after the last dose, are unwilling or unable to use FDA-approved forms of contraception. Targeting HIV-positive individuals. This category includes those who test positive for HBV or HCV infection using HBVeAg / DNA [positive anti-HCV (EIA) and confirmed HCV RIBA]. Subjects with latent or active TB. Subjects must have a negative Quantiferon TB Gold test result. Recent recipients of any approved or research-in-progress attenuated live vaccine within two months of a screening visit (including, but not limited to, any of the following: adenovirus [live oral adenovirus vaccine type 7], varicella [Varivax], hepatitis A [VAQTA], rotavirus [Rotashield], yellow fever [YF-Vax], measles and mumps [live measles and mumps virus vaccine], measles, mumps, and rubella vaccine [MMR-II], Sabin oral polio vaccine, and rabies vaccine [IMOVAX Rabies ID, RabAvert]). ANC<2000, platelet count<100×10 3 A result of a significantly abnormal general serum screening test, defined as / ml, SGOT or SGPT > 1.5 × upper limit of normal. Individuals deemed unable to follow the protocol. Subjects with active CMV or EBV infection, defined by CMV-specific serology (IgG or IgM) and confirmed by quantitative PCR, who either have or do not have the disease (quantitative PCR cutoff defined as having >50 copies of CMV or EBV DNA / PCR). Use of the study drug within 4 weeks of participation. A history or activity of inflammatory bowel disease, diverticular disease, or gastrointestinal perforation. Recent infection (within the past 6 weeks of screening) requiring the use of any antibody (oral, parenteral, or topical). Current or past (within the last 5 years) malignant tumors, excluding basal cell carcinoma, squamous cell carcinoma of the skin that has been completely excised, or non-recurrent (within the last 5 years) cervical intraepithelial neoplasia.

[0124] The applicant also conducted a study evaluating the cost-benefit analysis of desensitization compared to dialysis. The costs associated with transplantation after desensitization, including the cost of obtaining all medications and organs, treating rejection, and returning to dialysis after allograft loss, were favorable compared to the costs of continuing dialysis over the same period. Most importantly, in this cohort, transplantation provided a survival benefit. At 3 years, the mortality rate for patients who underwent desensitization and transplantation was 3.5%, compared to 22.8% for patients who continued dialysis.

[0125] If rejection (confirmed by biopsy) occurred during the study, patients were treated with pulsed methylprednisolone (10 mg / kg / day, up to 1000 mg for >100 kg, for 3 days) and antithymocyte globulin (1.5 mg / kg once daily for 4 days) for cell-mediated rejection that did not respond to pulsed steroids. Patients who experienced recurrent antibody-mediated rejection (ABMR) after treatment with the study drug were initially given pulsed methylprednisolone (10 mg / kg / day, up to 1000 mg for >100 kg) IV once daily for 3 doses, followed by, depending on the severity, IVIG 10% solution 2 mg / kg (up to 140 g for >70 kg) IV once, followed by rituximab (375 mg / m²). 2A single dose of IV was administered. If a rapid decline in the function of the allogeneic graft was observed and / or thrombotic microangiopathy was diagnosed, the patient underwent plasmapheresis 3 to 5 times, followed by weekly administration of anti-C5 (Eculizumab®) IV for 4 weeks (1200 mg in the first week, followed by 900 mg / week for a further 3 weeks). The efficacy of the treatment was evaluated by determining improvement in renal function and monitoring the DSA response, and allogeneic graft biopsies were repeated as needed. For the purposes of this study, ABMR was defined as follows: reduced function of allograft in high-risk transplant recipients (i.e., sensitized patients with a history of DSA) as measured by serum Cr and eGFR (defined as a >30% decrease from baseline); association with the presence of DSA (usually increasing in intensity) as measured by LUMINEX technology; and biopsy evidence based on BANFF 2015 classification (e.g., capillary vasculitis, inflammation, and C4d deposition).

[0126] Adverse events (AEs) and serious adverse events were monitored after treatment with crazakizumab. These included careful monitoring for potential infectious complications associated with crazakizumab treatment. Our group evaluated infectious complications associated with IVIG + rituximab desensitization and alemtuzumab induction therapy, as well as subsequent maintenance therapy with tacrolimus, MMF, and prednisone. The use of this desensitization protocol and subsequent alemtuzumab induction did not increase the risk of common or serious post-transplant infections compared to patients in the low-risk group. Serious infections were defined as any viral infection, and fungal or bacterial infections requiring antibiotics with IV or hospitalization. Therefore, the risk of infection in the study group (crazakizumab) after ABMR treatment is likely to be similar to and equivalent to that in unsensitized patients. All patients participating in this study required vaccination against Streptococcus pneumoniae.

[0127] In this study, all study patients, regardless of their cytomegalovirus (CMV) status, received ganciclovir intravenously during hospitalization and valganciclovir intravenously on an outpatient basis for 6 months post-kidney transplantation, with dose adjustments based on renal function. Fungal prophylaxis was achieved with fluconazole 100 mg once daily for 1 month post-transplantation. Prophylaxis against Pneumocystis jirovecii pneumonia and bacterial infections was achieved with trimethoprim 80 mg and sulfamethoxazole 400 mg once daily for 12 months post-transplantation. Viral polymerase chain reaction assays for CMV, Epstein-Barr virus, parvovirus B-19, polyomavirus BK, and JC were performed on study patients once a month for 6 months post-transplantation.

[0128] Various embodiments of the present invention are described above in the detailed description. While these descriptions directly illustrate the embodiments described above, it will be understood that those skilled in the art may conceive of modifications and / or variations to the specific embodiments shown and described herein. Any such modifications or variations that fall within the scope of this description are also intended to be included herein. Unless otherwise specified, it is the inventors' intention that words and phrases in this specification and in the claims have their usual and idiomatic meanings to those skilled in the appropriate art.

[0129] The above description of various embodiments of the Invention known to the applicant at the time of filing of this application is presented for illustrative and explanatory purposes, and is intended for this purpose. This specification is not intended to be exhaustive or to limit the Invention to the detailed forms disclosed, and many modifications and variations are possible in light of the above teachings. The embodiments described are useful in illustrating the principles of the Invention and its practical applications, and enable those skilled in the art to utilize the Invention in various embodiments and with various modifications suitable for specific applications that can be conceivable. Accordingly, it is intended that the Invention is not limited to the specific embodiments disclosed for carrying out the Invention.

[0130] While specific embodiments of the present invention are shown and described herein, modifications and alterations may be made based on the teachings herein without departing from the present invention and its broader aspects, and it will be apparent to those skilled in the art that the appended claims encompass all modifications and alterations that are in the true spirit and scope of the present invention. In general, it will be understood to those skilled in the art that the terms used herein are generally intended to be “open” terms (for example, “including” should be interpreted as “including but not limited to,” “having” should be interpreted as “having at least,” and “include” should be interpreted as “including but not limited to,” etc.).

[0131] Where used herein, the terms “comprising” or “comprise” are used in reference to compositions, methods, and their respective components that are useful to a particular implementer form, and are also open to the inclusion of unspecified elements, whether useful or not. In general, it will be understood by those skilled in the art that the terms used herein are generally intended to be “open” terms (for example, “including” should be interpreted as “including but not limited to,” “having” should be interpreted as “having at least,” and “include” should be interpreted as “including but not limited to”). In this specification, the open-ended term “comprising” is used as a synonym for terms such as including, containing, or having, but the invention or its embodiments may be described using alternative terms such as “consisting of” or “essentially consisting of.”

[0132] Unless otherwise specified, all numbers representing quantities should be understood to be modified in all cases by the term “approximately.” The term “approximately” can refer to ±10% of the value mentioned. Where specifically defined and stated in the claims, the term “approximately” can refer to ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1% of the value mentioned; for example, the claims may state that the value is approximately X, and therefore approximately ±6%.

[0133] Where a range of values ​​is given, the range between the upper and lower limits of that range, and each number including these, is intended to be disclosed herein. It should be understood that any numerical range enumerated herein is intended to include all subranges that fall within that range. For example, the range "1 to 10" is intended to include the range between the enumerated minimum value of 1 and the enumerated maximum value of 10, and all subranges including these; that is, it has a minimum value of 1 or more and a maximum value of 10 or less. Since the numerical ranges in this disclosure are continuous, all values ​​between the minimum and maximum values ​​are included.

Claims

1. For use in methods for reducing and / or eliminating donor-specific antibodies in human leukocyte antigen (HLA) sensitized subjects, Clazakizumab; interleukin-6 (IL-6) binding fragment of clzakizumab; or V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide as contained in SEQ ID NOs: 1, 2 or 3, and 4, respectively. H A polypeptide and V containing the CDR1 polypeptide, CDR2 polypeptide, and CDR3 polypeptide contained in SEQ ID NOs. 5, 6, and 7, respectively. L polypeptide having polypeptide, A pharmaceutical composition containing, The method includes administering an effective amount of the pharmaceutical composition to a target, The subjects in question are those who require or have received solid organ transplants. The aforementioned pharmaceutical composition.

2. The pharmaceutical composition according to claim 1, further comprising one or more pharmaceutically acceptable excipients.

3. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered before solid organ transplantation.

4. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered after solid organ transplantation, during solid organ transplantation, or both.

5. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered both before and after solid organ transplantation.

6. The pharmaceutical composition according to claim 1 or 2, further comprising administering a standard therapeutic procedure including intravenous immunoglobulin (IVIG), rituximab, plasma exchange, or a combination thereof.

7. The pharmaceutical composition according to claim 6, wherein a standard treatment procedure is administered before the pharmaceutical composition.

8. The pharmaceutical composition according to claim 1 or 2, wherein the solid organ is a kidney.

9. The pharmaceutical composition according to claim 1 or 2, wherein the solid organ is one or more of the heart, liver, lungs, pancreas, and intestines.

10. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered subcutaneously or intravenously.

11. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered subcutaneously over a period of at least one month and up to eighteen months at an average dose of approximately 0.1 to 1 mg / month, 1 to 5 mg / month, 5 to 10 mg / month, 10 to 20 mg / month, 20 to 30 mg / month, or 30 to 40 mg / month.

12. The pharmaceutical composition according to claim 1 or 2, wherein multiple doses of the pharmaceutical composition are administered at intervals of approximately once a month over a period of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months.

13. The pharmaceutical composition according to claim 1 or 2, wherein the pharmaceutical composition is administered subcutaneously in an average dose of approximately 10 to 30 mg / dose in one, two, three, four, five, or six doses before solid organ transplantation, and in one, two, three, four, five, six, seven, eight, nine, ten, eleven, or twelve doses after solid organ transplantation.

14. The pharmaceutical composition according to claim 1 or 2, wherein the target is a human.

15. The method further comprises administering one or more anti-infective agents to a target, according to claim 1 or 2.

16. The pharmaceutical composition according to claim 15, wherein one or more anti-infective agents are administered together with or after solid organ transplantation.

17. The pharmaceutical composition according to claim 14, wherein the antiinfective agent comprises ganciclovir, valganciclovir, fluconazole, trimethoprim, sulfamethoxazole, or a combination thereof.

18. The pharmaceutical composition according to claim 1 or 2, further comprising administering a standard treatment including intravenous immunoglobulin (IVIG), rituximab, plasma exchange, or a combination thereof; an anti-infective agent; or a combination of a standard treatment and an anti-infective agent.

19. The method further comprises selecting a human subject in which the calculated panel-reactive antibody (cPRA) is 50% or more, or performing a panel-reactive antibody assay, and determining a human subject in which the cPRA is 50% or more, according to claim 1 or 2.

20. The pharmaceutical composition according to claim 1 or 2, wherein, after solid organ transplantation, the subject shows no detectable evidence of antibody-mediated rejection of the solid organ transplant, no detectable evidence of viral infection, or both.

21. The method further comprises administering an antibody induction therapy containing alemtuzumab, anti-thymocyte globulin, or both, after solid organ transplantation; administering an immunosuppressive therapy containing tacrolimus, mycophenolate mofetil, prednisone, or a combination thereof; or administering an antibody induction therapy and an immunosuppressive therapy, according to claim 1 or 2.

22. The pharmaceutical composition according to claim 1 or 2, wherein the subject requiring solid organ transplantation is either undergoing a solid organ transplantation or has previously undergone a solid organ transplantation, and the method further comprises performing one or more immunomonitoring tests on the subject, the immunomonitoring tests comprising assaying a blood sample of the subject to quantify the levels of markers including CRP, Treg, Tfh, Th17, B cells, IL-6, plasma cells, plasmablast IgG, or a combination thereof.

23. If immune monitoring shows improvement based on one or more markers that are at a lower level compared to baseline measurements obtained at or before solid organ transplantation, or based on one or more markers that are at a lower level compared to those obtained from past immune monitoring, further administration of the pharmaceutical composition is discontinued or limited to a further six months or less; if immune monitoring shows insufficient results based on markers that are at the same level or higher than baseline measurements or those obtained from past immune monitoring, one or more doses of the pharmaceutical composition are administered, according to claim 22.

24. The pharmaceutical composition according to claim 1 or 2, wherein the subject requiring solid organ transplantation is either undergoing a solid organ transplantation or has previously undergone a solid organ transplantation, and the method further comprises measuring the amount of glomerular filtration rate, DSA, or both after solid organ transplantation.

25. The pharmaceutical composition according to claim 24, wherein if the amount of glomerular filtration rate, DSA, or both is similar to or lower than baseline levels measured before or at the time of solid organ transplantation, further administration of the pharmaceutical composition is discontinued or limited to a further six months or less; if the amount of glomerular filtration rate, DSA, or both is higher than baseline levels, one or more doses of the pharmaceutical composition are administered.

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