Combination medicine containing anti-CD205 antibody and immune checkpoint inhibitor

Abstract

The present invention relates to methods for increasing anti-tumor immune responses in patients suffering from cancer, methods for treating or preventing cancer, and methods for enhancing the effect of inhibitors of PD1 / PD-L1 interaction, as well as pharmaceutical combinations comprising (a) an antibody, or antigen-binding portion thereof, targeting CD205, and (b) a PD1 / PD-L1 checkpoint inhibitor.

Description

[Technical field]

[0001] preface The present disclosure relates generally to the fields of immunology and molecular biology. More specifically, the present application provides methods for increasing anti-tumor immunity, and more specifically T-cell mediated tumor specific responses, or the number of T-cells, in patients suffering from cancer, methods for treating or preventing cancer, and methods for enhancing the effect of inhibitors of PD1 / PD-L1 interaction. Also provided is a combination drug comprising (a) an antibody, or an antigen-binding portion thereof, targeting CD205, and (b) a PD1 / PD-L1 checkpoint inhibitor. [Background technology]

[0002] Dendritic cells (DCs) play an important role in initiating immune responses against both foreign and endogenous antigens. There are two types of DCs with different origins and functions: myeloid dendritic cells (mDCs) and plasmacytoid dendritic cells (pDCs). Both mDCs and pDCs can efficiently induce CD4+ and CD8+ T cell responses against pathogens, and both can also interact with natural killer (NK) cells. CD4+, CD8+ and NK cells play an important role in immune-mediated anti-cancer responses. However, pDCs as well as mDCs can also induce tolerance against cancer by inducing Regulatory T cells (Tregs) (Ito et al. JEM,

[2007] ), which in turn blocks T cell proliferation and T cell activation.

[0003] Liu, X et al (Journal of Cancer,

[2019] , Vol. 10, p 6711-6715) disclose that Tregs and pDCs are the main immunosuppressive cells in the tumor microenvironment in gastric cancer. Liu, X et al show that patients with higher pDC numbers in both gastric cancer tissue and peripheral blood have shorter overall survival than patients with lower pDC numbers in each compartment. Similar adverse effects of DCs in cancer tissue on cancer patient survival have been reported in breast, ovarian and renal cancer.

[0004] DCs use CD205 (also known as DEC205 and lymphocyte antigen 75) as an endocytic receptor for the presentation of self- and foreign antigens to induce either immune responses or tolerance. CD205 is expressed on both CD8+ mDCs and CD8+ pDCs (Shrimpton et al., 2009). CD205 distinguishes two major types of DCs: CD8+ / CD205+ DCs reside in the T cell zone of lymphoid organs, and CD8- / 33D1+ DCs reside in the red pulp and marginal zone (Dudziak et al. Science Vol. 315 p107-111

[2007] ). CD8+ CD205+ DCs have been reported to selectively induce immunosuppressive Tregs (Yamazaki et al., 2008; Okeke and Uzonna, 2019; Simon and Bromberg, 2016; Kushwah and Hu, 2011), and the generation of Treg cells in blood has been associated with the ratio of CD8+ CD205+ DCs among all CD11c+ DCs (Simon and Bromberg, 2016). Tregs are known to suppress tumor CD8+ or specific cytotoxic T cells (Chen et al. 2005; Li et al. 2020).

[0005] WO2009 / 061996 discloses isolated monoclonal antibodies that bind to human CD205 and related antibody-based compositions and molecules. Pharmaceutical compositions containing the antibodies, as well as therapeutic and diagnostic methods for using the antibodies, are also disclosed.

[0006] WO2008 / 104806 discloses affinity reagents capable of binding to CD205 for use in treating or preventing cancer.

[0007] WO2015 / 052537 discloses specific isolated antibodies capable of binding to CD205 and their use in treating various cancers.

[0008] Programmed cell death 1 (PD1) and programmed cell death ligand 1 (PD-L1) are immune checkpoint proteins whose interaction plays a key role in limiting the activity of T cells and thus constitutes a major immune resistance mechanism by which tumor cells escape immune surveillance. is provided.

[0009] A number of agents directed against the PD-1 / PD-L1 pathway have been developed and have been shown to be effective in treating many cancer types.

[0010] In recent years, numerous clinical trials have been conducted combining PD1 / PD-L1 checkpoint inhibitors with a wide range of additional agents. The majority of these have combined PD1 with CTLA4, antiangiogenic agents or chemotherapeutic agents. The results of these trials have shown mixed results (Schmidt, EV, Semin Immunopathol; 41(1), 21-30

[2019] ).

[0011] Gastric cancer is one of the most common malignant tumors of the digestive system and one of the top five malignant tumors in terms of incidence and mortality. Advanced gastric cancer is currently limited to treatment options with chemotherapy as the first line treatment. Trastuzumab and ramucirumab are also approved for HER-2 and VEGF positive tumors, respectively, that have failed first line treatment. The overall survival rate for gastric cancer worldwide is only ~20%. Immune checkpoint inhibitors alone have shown some efficacy against gastric cancer but have shown poor efficiency (Song, X., et al. Oncology letters, 20(4),

[2020] ).

[0012] Endometrial cancer is the most common gynecologic cancer in the United States, with approximately 50,000 women diagnosed each year. Advanced endometrial cancer is currently treated with radiation therapy, chemotherapy, or hormonal therapy. However, there is a need to develop new targeted therapies to treat refractory or recurrent disease. Summary of the Invention [Problem to be solved by the invention]

[0013] Summary of the Invention The present invention is based on the surprising discovery by the present inventors that cancer patients with a reduced population of CD205+ immunoregulatory cells exhibit a significant increase in the number of both CD4+ and CD8+ T-cells in their peripheral blood. The present inventors have also identified that, together with this increase in the number of T-cells, there is also a significant increase in the number of both CD4+ and CD8+ T-cells expressing PD1.

[0014] We also observed that the absolute number of pDCs present in patient blood samples initially declined rapidly after treatment with CD205-DM4 antibody drug conjugate (ADC), then increased again, doubling by day 21 after treatment. The same pattern was seen with CD205+ pDC cells. Similarly, the same pattern was seen with CD205+ mDC cells, which declined until day 8 after treatment with CD205-DM4 ADC, but then quadrupled by day 21.

[0015] The inventors believe that the reduction in CD205+ immunoregulatory cells and the subsequent increase in CD4+ and CD8+ T-cells enhances the patient's immune response against the tumor. The inventors further hypothesize that the T-cells are activated after the reduction in CD205+ immunoregulatory cells, with an increase in the number of CD4+ and CD8+ T-cells. The inventors also hypothesize that the observed reduction in the CD205+ pDC population will reverse immune suppression in patients treated with CD205-DM4 ADC. This is supported by the disclosure of Liu et al., which suggests that pDCs are the major immunosuppressive cells in the tumor microenvironment of gastric cancer and are associated with shorter overall survival, as discussed above. With a significant increase in PD1 / PD-L1 expression, the enhancement of the immune response can be prolonged by administering immune checkpoint inhibitors, thereby avoiding immune suppression. [Means for solving the problem]

[0016] According to a first aspect of the present invention there is provided a method for treating or preventing cancer, said method comprising the step of administering to a patient in need thereof a therapeutically effective amount of an antibody or antigen binding fragment thereof which modulates a population of CD205+ immunoregulatory cells, and a therapeutically effective amount of a composition comprising a checkpoint inhibitor, is provided.

[0017] It will be apparent to those skilled in the art that the antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells and the composition comprising the checkpoint inhibitor can be administered simultaneously, separately or sequentially, preferably sequentially.

[0018] In one embodiment, the checkpoint inhibitor is PD1, PD-L1, PD-L2, CTLA-4, ICOS, TIGIT, CD28, TMIGD2, CD137, CD137L, CD27, OX40, OX40L, LAG3, VISTA, GITR, DNAM-1, CD96, 2B4, TIM-3, CEACAM, CRTAM, SLAMF6, Galectin-9, CD48, CD155, GITRL, CD40, CD40L, CD70, HVEM, B7-H7, B7-H3, B7-H4, ICOSL, CD80, CD86, BTLA, CD160, LIGHT, adenosine A2a receptor, SIRP alpha, DC-SIGN, CD200R, DR3, TL1A, CD200, The antibody targets a checkpoint protein selected from the group including BTN2A1, CD47, IDO, and TDO.

[0019] In one embodiment, the checkpoint inhibitor is PD1 or PD-L1, preferably PD1.

[0020] According to a second aspect of the invention there is provided a method for enhancing the effect of an inhibitor of PD1 / PD-L1 in a patient, said patient having been identified as in need thereof, said method comprising the steps of administering to said patient (a) a therapeutically effective amount of an antibody or antigen-binding fragment thereof which modulates a population of CD205+ immunoregulatory cells, and (b) a composition comprising an inhibitor of PD1 / PD-L1 interaction. is provided.

[0021] It will be apparent to one of skill in the art that the composition comprising an antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells and an inhibitor of PD1 / PD-L1 interaction can be administered simultaneously, separately or sequentially, preferably sequentially.

[0022] It will be readily apparent to one skilled in the art that the term enhance, as used in this context, means increasing the level of effect of the immune checkpoint inhibitor, such that after modulating the population of CD205+ immunoregulatory cells, a higher level of cytotoxicity is observed than before it is decreased, or the time period during which the immune checkpoint inhibitor is effective is increased. Administering an antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells can be considered to act to stimulate the immune system to express immune checkpoint proteins. Thus, administration of an immune checkpoint inhibitor results in higher and / or longer lasting cytotoxicity.

[0023] According to a third aspect of the present invention there is provided a method for increasing an anti-tumor immune response in a patient suffering from cancer, said method comprising the step of administering to said patient a therapeutically effective amount of an antibody or antigen binding fragment thereof which modulates a population of CD205+ immunoregulatory cells. is provided.

[0024] As used in the context of the third embodiment, the term "increasing an anti-tumor immune response" means: This means that when measured as an increase in the number of immune cells present in the patient, there is a greater immune response against the cancer after the reduction in CD205+ immunoregulatory cells than before the reduction.

[0025] In one embodiment, the anti-tumor immune response is an immune cell-mediated tumor-specific response. In a preferred embodiment, the immune response is a T-cell-mediated tumor-specific response.

[0026] In a further embodiment, the anti-tumor immune response is a NK cell-mediated tumor-specific response.

[0027] According to a fourth aspect of the present invention there is provided a method for increasing the number of T-cells in a patient suffering from cancer, said method comprising the step of administering to said patient an antibody or antigen binding fragment thereof which modulates the population of CD205+ immunoregulatory cells. is provided.

[0028] In one embodiment, the T-cells are CD8+ T-cells.

[0029] In another embodiment, the T-cells are CD4+ T-cells.

[0030] In a further embodiment, the T-cells are tumor-specific T-cells.

[0031] According to a further aspect, there is provided a method for reducing tumor size in a patient suffering from cancer, said method comprising administering to said patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that modulates a population of CD205+ immunoregulatory cells. is provided.

[0032] In one embodiment, the tumor is a metastatic tumor. In a further embodiment, the metastatic tumor is in the lung or liver.

[0033] For the avoidance of doubt, any embodiment of the invention described below will, where appropriate, refer to all earlier aspects of the invention.

[0034] In one embodiment of the invention, the CD205+ immunoregulatory cells are CD8+. Preferably, the CD205+ CD8+ immunoregulatory cells are decreased.

[0035] In one embodiment, the immunoregulatory cells are T-Reg cells.

[0036] In one embodiment of the invention, the CD205+ immunoregulatory cells are pDCs and / or mDCs. Preferably, the number of pDCs and / or mDCs is increased.

[0037] In one embodiment of the invention, the CD205+ immunoregulatory cells are CD4+. Preferably, the CD205+ CD4+ immunoregulatory cells are decreased.

[0038] In one embodiment, the immunoregulatory cells are T-Reg cells.

[0039] In one embodiment of the invention, the immunomodulatory cells are immunosuppressive cells.

[0040] In some embodiments, the immunoregulatory cells are dendritic cells.

[0041] In one embodiment of the invention, the patient receives a cancer vaccine simultaneously, separately, sequentially or subsequently.

[0042] In a further embodiment of the invention, the patient is administered simultaneously, separately, sequentially or subsequently a bispecific antibody. Preferably, said bispecific antibody is a T-cell engager (BiTE). More preferably, said bispecific antibody comprises a first binding domain that binds to CD3.

[0043] Preferably, the bispecific antibody comprises a second binding domain that binds to a tumor-specific antigen.

[0044] In one embodiment, the patient is one who is refractory to at least one chemotherapy or whose cancer has progressed during at least one chemotherapy.

[0045] In another embodiment, the patient is refractory to checkpoint modulator therapy.

[0046] In a further embodiment, the patient is ineligible for checkpoint modulator therapy.

[0047] One of skill in the art will appreciate that patients who are ineligible for checkpoint modulator therapy are patients who meet the criteria specified for treatment of a particular indication.

[0048] In one embodiment, the checkpoint modulator therapy is PD1 therapy.

[0049] In a further embodiment, the patient has a cancer that is PDL1 negative or low.

[0050] One of skill in the art will understand that the term low PDL1 expression refers to cancers that have PD-L1 expression less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%.

[0051] As used in this application, the term PDL1-negative means a cancer that has no detectable PDL1 expression using IHC.

[0052] In a further embodiment, the cancer is MSI stable.

[0053] In one embodiment, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or more of the CD8+ cells in a blood sample previously isolated from the patient are CD205+.

[0054] In another embodiment, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or more of the CD4+ cells in a blood sample previously isolated from the patient are CD205+.

[0055] In further embodiments, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, or more of the pDCs and / or mDCs in a blood sample previously isolated from the patient are CD205+.

[0056] In one embodiment, the antibody, or antigen-binding portion thereof, binds to CD205.

[0057] In one preferred embodiment, an antibody, or antigen-binding portion thereof, that binds CD205 for use in the methods of the invention comprises: A heavy chain variable region comprising: i) a first vhCDR comprising SEQ ID NO: 5; ii) a second vhCDR comprising SEQ ID NO: 6; and iii) a third vhCDR comprising SEQ ID NO: 7; and A light chain variable region comprising: i) a first vlCDR comprising SEQ ID NO: 8; ii) a second vlCDR comprising SEQ ID NO: 9; and iii) a third vlCDR comprising SEQ ID NO: 10; Optionally, wherein any one or more of the above SEQ ID NOs independently contain one, two, three, four, or five amino acid substitutions, additions, or deletions.

[0058] In one embodiment, the antibody is internalized.

[0059] In a further embodiment, the antibody, or antigen-binding portion thereof, for use in the methods of the invention comprises a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO:1, and a light chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO:2.

[0060] Heavy and light chain variable regions having sequence identity in ranges intermediate to the above values, for example, at least 80-85%, 85-90%, 90-95% or 95-100% to any of the above sequences, are also intended to be encompassed by the invention.

[0061] In one embodiment, an anti-CD205 antibody, or antigen-binding portion thereof, for use in the methods of the invention comprises the CDR1, CDR2, and CDR3 domains of the heavy chain variable (VH) region of an anti-CD205 antibody having the sequence set forth in SEQ ID NO:1, and / or the CDR1, CDR2, and CDR3 domains of the light chain variable (VL) region of an anti-CD205 antibody having the sequence set forth in SEQ ID NO:2.

[0062] In a preferred embodiment, the CDRs are defined according to the Kabat or Chothia system.

[0063] In another embodiment, an antibody, or antigen binding portion thereof, for use in the methods of the invention comprises a heavy chain variable region comprising a first vhCDR comprising SEQ ID NO:5; a second vhCDR comprising SEQ ID NO:6; and a third vhCDR comprising SEQ ID NO:7; and a light chain variable region comprising a first vlCDR comprising SEQ ID NO:8; a second vlCDR comprising SEQ ID NO:9; and a third vlCDR comprising SEQ ID NO:10.

[0064] In another embodiment, an anti-CD205 antibody, or antigen binding portion thereof, for use in the methods of the invention binds to human CD205 and comprises a heavy chain variable region comprising SEQ ID NO:1 and / or conservative sequence modifications thereof. The antibody may further comprise a light chain variable region comprising SEQ ID NO:2 and / or conservative sequence modifications thereof.

[0065] In another embodiment, an anti-CD205 antibody, or antigen-binding portion thereof, for use in the methods of the invention comprises a heavy chain scaffold region comprising an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to the scaffold of the heavy chain variable region of SEQ ID NO: 1, as set forth in SEQ ID NOs: 12, 13, 14, and 15. In another embodiment, an anti-CD205 antibody, or antigen-binding portion thereof, for use in the methods of the invention comprises a light chain framework region comprising an amino acid sequence that is at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to the framework of the light chain variable region of SEQ ID NO:2, as set forth in SEQ ID NOs:16, 17, 18, and 19.

[0066] In one embodiment, an anti-CD205 antibody, or antigen-binding portion thereof, for use in the methods of the invention comprises a heavy chain variable region and a light chain variable region, respectively, encoded by a nucleic acid sequence comprising SEQ ID NOs: 3 and 4, or a nucleic acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the above nucleic acid sequences, or a sequence that differs from SEQ ID NOs: 3 and 4 due to degeneracy of the genetic code.

[0067] In one embodiment, the antibody or antigen-binding portion thereof for use in the methods of the invention further comprises a covalently-linked moiety. Preferably, the moiety is a drug. More preferably, the drug is selected from the group consisting of maytansinoids, dolastatins, hemiasterlins, auristatins, trichothecenes, calicheamicins, duocarmycins, bacterial immunotoxins, pyranoindoidinoquinolines, camptothecins, anthracyclines, antheamicins, thienoindoles, amatoxins, CC1065, or taxol, and derivatives thereof.

[0068] In a preferred embodiment, the drug is a maytansinoid selected from the group consisting of DM4 and DM1, preferably DM4.

[0069] In one embodiment, the cancer is a CD205 positive cancer.

[0070] In a preferred embodiment, a composition that modulates the population of CD205+ immunoregulatory cells for use in the methods of the invention comprises an antibody that binds to CD205, including: A heavy chain variable region comprising: i) a first vhCDR comprising SEQ ID NO: 5; ii) a second vhCDR comprising SEQ ID NO: 6; and iii) a third vhCDR comprising SEQ ID NO: 7; and A light chain variable region comprising: i) a first vlCDR comprising SEQ ID NO: 8; ii) a second vlCDR comprising SEQ ID NO: 9; and iii) a third vlCDR comprising SEQ ID NO: 10; Here, the antibody is conjugated to a cytotoxic moiety that comprises the maytansinoid DM4.

[0071] In one embodiment, the PD1 / PD-L1 inhibitor is an antibody.

[0072] A person skilled in the art will appreciate that the PD1 / PD-L1 antibody may be any suitable antibody.

[0073] In preferred embodiments, the anti-PD-1 antibody for use in the methods of the invention is selected from the group including: nivolumab (MDX-1 106, Opdivo; Bristol-Myers Squibb), pembrolizumab (MK-3475, Keytruda, lambrolizumab, BMS-936558; Merck), dostallimab (TSR-042, Tesaro, Inc.), cemiplimab (REGN2810, Libtayo; Regeneron Pharmaceuticals), EH12.2H7 (BioLegend, catalog number 329902), Balstilimab (Agenus Inc.).

[0074] In other preferred embodiments, the anti-PD-1 antibody for use in the methods of the invention is selected from the group including: avelumab (Bavencio; EMD Serono, Pfizer), durvalumab (Imfinzi, AstraZeneca), BMS-936559, atezolizumab (Tecentriq, Genentech).

[0075] In one embodiment of the invention, the checkpoint inhibitor is administered between 7 days and 12 weeks, preferably between 7 days and 10 weeks, or 7 days and 8 weeks, or 7 days and 6 weeks, or 7 days and 4 weeks, or 7 days and 21 days, or 10 days and 19 days, 12 days and 16 days, 14 days and 16 days, or 19 days and 28 days, more preferably between 20 days and 25 days, and most preferably between 21 days and 24 days, after administration of an antibody or antigen-binding portion thereof that binds CD205.

[0076] In one embodiment of the invention, the checkpoint inhibitor is administered 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks after administration of the antibody.

[0077] One skilled in the art will appreciate that as the number or proportion of T-cells expressing PD1 increases, the immune response is suppressed, possibly via PD1 / PD-L1 immune checkpoint interactions. If these interactions are disrupted, for example by administering a checkpoint inhibitor, the patient's immune response to tumors may persist and T-cells may become more cytotoxic against the tumor.

[0078] In one embodiment, the patient receives at least one cycle, at least two cycles, at least three cycles, at least four cycles, or at least five cycles of an antibody or antigen-binding fragment thereof that modulates a population of CD205+ immunoregulatory cells prior to administration of the checkpoint modulator.

[0079] In another embodiment, the patient receives 1 to 5 cycles, 2 to 4 cycles, or 2 to 3 cycles of administration of an antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells prior to administration of the checkpoint modulator.

[0080] In some embodiments, the cancer is pancreatic cancer, ovarian cancer, breast cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastroesophageal junction cancer, skin cancer, thyroid cancer, lung cancer, kidney cancer, liver cancer, head and neck cancer, bladder cancer, gastric cancer, and malignant leukemia, leukemia, preferably acute myeloid leukemia or chronic lymphocytic leukemia, myeloma, preferably multiple myeloma and lymphoma, preferably diffuse large B-cell lymphoma (DLBCL), B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, lymphoma of mucosa-associated lymphoid tissue (MALT), T-cell / histiocyte-rich B-cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma, small lymphocytic lymphoma, marginal zone lymphoma, T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma and angioimmunoblastic T-cell lymphoma.

[0081] Preferably, the cancer is selected from the group comprising: gastric cancer, endometrial cancer, gastroesophageal junction cancer, esophageal cancer, ovarian cancer, lung cancer, breast cancer, renal cancer, and bladder cancer. Most preferably, the cancer is gastric cancer.

[0082] In one embodiment, the breast cancer is triple negative breast cancer (TNBC). In another embodiment, the breast cancer is Her2-ve breast cancer.

[0083] In one embodiment, administration of the anti-CD205 antibody, or antigen-binding portion thereof, increases the number of CD8+ T-cells in the patient, resulting in increased T-cell cytotoxicity against the tumor.

[0084] In a preferred embodiment, the patient according to any of the above aspects is a human.

[0085] In some embodiments of the invention, the antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells is an anti-CD205-DM4 ADC.

[0086] In one embodiment, the anti-CD205-DM4 ADC is administered to the patient at a dosage range of about 0.8 to 10 mg / kg, e.g., 1.0 mg / kg to 8.0 mg / kg, 1.2 mg / kg to 7.5 mg / kg, 1.4 mg / kg to 7.0 mg / kg, 1.6 to 6.0 mg / kg, 1.6 to 5 mg / kg, 2.0 to 4 mg / kg, 2.5 to 3.6 mg / kg of host body weight. For example, dosages can be 0.8 mg / kg, 1.0 mg / kg, 1.2 mg / kg, 1.4 mg / kg, 1.6 mg / kg body weight, 2.0 mg / kg body weight, 2.5 mg / kg body weight, 3.5 mg / kg body weight, 4 mg / kg body weight, or 5 mg / kg body weight. Most preferably, the dosage is 3.5 mg / kg. Exemplary treatment regimens entail administration once per week, once per two weeks, once per three weeks, once per four weeks, once per month, once per six weeks, once per three months, or once per three to six months.

[0087] Preferred dosing regimens of anti-CD205-DM4 ADC for use in the methods of the invention include 2.0 mg / kg body weight, 2.5 mg / kg body weight, 3.0 mg / kg body weight, 3.5 mg / kg body weight, or 5 mg / kg body weight via intravenous administration, with the antibody drug conjugate administered using one of the following dosing schedules: (i) once every 3 weeks for 6 doses; (ii) once every 3 weeks; (iii) 2.5 mg / kg body weight once followed by 2 mg / kg body weight every 3 weeks.

[0088] Further preferred dosing regimens of anti-CD205 antibody drug conjugates for use in the methods of the invention include 0.8 mg / kg body weight, 1.0 mg / kg body weight, 1.2 mg / kg body weight, or 1.4 mg / kg body weight via intravenous administration, wherein the antibody drug conjugate is administered using one of the following dosing schedules: (i) once per week; (ii) once per week for four doses; (iii) once per week for three doses; (iv) three times per week once every three weeks.

[0089] In one embodiment, the PD1 antibody is administered to the patient at a dosage range of 200 mg to 480 mg, e.g., 200 mg, 240 mg, 400 mg, or 480 mg. Exemplary treatment regimens entail administration once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, or once every 6 weeks.

[0090] In another embodiment, for administration of the PD-L1 antibody, the dosage ranges from 800 mg to 1500 mg, e.g., 800 mg, 1200 mg, or 1500 mg. Exemplary treatment regimens entail administration once every two weeks, once every three weeks, or once every four weeks.

[0091] According to a further aspect of the invention, a pharmaceutical combination comprising: a) an anti-CD205 antibody or an antigen-binding portion thereof, wherein said antibody comprises: A heavy chain variable region comprising: i) a first vhCDR comprising SEQ ID NO: 5; ii) a second vhCDR comprising SEQ ID NO: 6; and iii) a third vhCDR comprising SEQ ID NO: 7; and A light chain variable region comprising: i) a first vlCDR comprising SEQ ID NO: 8; ii) a second vlCDR comprising SEQ ID NO: 9; and iii) a third vlCDR comprising SEQ ID NO: 10; and b) checkpoint inhibitors, is provided.

[0092] In one embodiment, the pharmaceutical combination is in the form of a combined preparation for simultaneous, separate or sequential use.

[0093] In a further embodiment, the checkpoint inhibitor is a PD1 / PD-L1 checkpoint inhibitor, preferably, the PD1 / PD-L1 checkpoint inhibitor is an antibody.

[0094] Preferably, the combination pharmaceutical is for the treatment of cancer.

[0095] In one embodiment, the PD1 / PD-L1 checkpoint inhibitor is an antibody.

[0096] A person skilled in the art will appreciate that the PD1 / PD-L1 antibody can be any suitable antibody.

[0097] In preferred embodiments, the anti-PD-1 antibody is selected from the group including: nivolumab (MDX-1 106, Opdivo; Bristol-Myers Squibb), pembrolizumab (MK-3475, Keytruda, lambrolizumab, BMS-936558; Merck), dostallimab (TSR-042, Tesaro, Inc.), cemiplimab (REGN-2810, Libtayo; Regeneron), EH12.2H7 (BioLegend, Cat. No. 329902).

[0098] In other preferred embodiments, the anti-PD-L1 antibody is selected from the group including: avelumab (Bavencio; EMD Serono, Pfizer), durvalumab (Imfinzi, AstraZeneca), BMS-936559, atezolizumab (Tecentriq, Genentech).

[0099] In further embodiments, the antibody, or antigen-binding portion thereof, comprises a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 1, and a light chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 2. In a preferred embodiment, the antibody, or antigen-binding portion thereof, comprises a heavy chain variable region having the sequence of SEQ ID NO: 1, and a light chain variable region having the sequence of SEQ ID NO: 2.

[0100] In a further embodiment, the antibody comprises a heavy chain having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 100, and a light chain having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 101. In a preferred embodiment, the antibody comprises a heavy chain having the sequence of SEQ ID NO: 100, and a light chain having the sequence of SEQ ID NO: 101.

[0101] All of the antibodies disclosed in this application can be full-length, e.g., any of the following isotypes: IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgAsec, IgD, and IgE. Alternatively, the antibodies can be fragments, e.g., antigen-binding portions or single chain antibodies (e.g., Fab, F(ab')2, Fv, single chain Fv fragments, isolated complementarity determining regions (CDRs), or combinations of two or more isolated CDRs). The antibodies can be any type of antibody, including, but not limited to, human, humanized, and chimeric antibodies.

[0102] In one embodiment, the anti-CD205 antibody, or antigen-binding portion thereof, further comprises a covalently-linked moiety. Preferably, the moiety is a drug. More preferably, the drug is selected from the group consisting of maytansinoids, dolastatins, hemiasterlins, auristatins, trichothecenes, calicheamicins, duocarmycins, bacterial immunotoxins, pyranoindoidinoquinolines, camptothecins, anthracyclines, antheamicins, thienoindoles, amatoxins, CC1065, or taxol, and derivatives thereof.

[0103] In a preferred embodiment, the drug is a maytansinoid selected from the group consisting of DM4 and DM1, preferably DM4.

[0104] In a further embodiment, the pharmaceutical combination comprises at least one pharma- ceutically acceptable diluent, excipient, or carrier.

[0105] A further aspect of the invention provides a composition or pharmaceutical combination of the invention for use in treating cancer. is provided.

[0106] There is also provided the use of components (a) and (b) as defined above in the manufacture of a combined medicament for separate, sequential use for the treatment of cancer.

[0107] According to a further aspect of the invention there is provided a method for selecting a patient suitable for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, wherein said patient is suffering from cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD8+ cells in a blood sample previously isolated from said patient are CD205+; and Administering to the patient a therapeutically effective amount of an anti-CD205 antibody or antigen-binding fragment thereof.

[0108] According to a further aspect of the invention there is provided an in vitro method for selecting a patient for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, said method comprising the steps of: a. determining the percentage of CD8+ cells in a blood sample previously isolated from said patient that are CD205+ cells; and b. selecting the patient for treatment with an antibody or antigen-binding fragment thereof that binds CD205 if at least 20% of the CD8+ cells in the blood sample are CD205+; is provided.

[0109] In one embodiment, the method for selecting a patient further comprises the step of administering to said patient a therapeutically effective amount of said antibody or antigen-binding fragment thereof that binds to CD205.

[0110] According to another aspect of the invention, there is provided a method for determining the efficacy of an antibody or antigen-binding fragment thereof that binds CD205 in treating cancer in a patient, said method comprising the steps of: obtaining a blood sample from the patient; identifying whether at least 20% of the CD8+ cells in the blood sample are CD205+; is provided.

[0111] In one embodiment, the method for determining efficacy further comprises administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds to CD205 if at least 20% of the CD8+ cells in the blood sample are CD205+.

[0112] In further embodiments, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD8+ cells are CD205+.

[0113] According to a further aspect of the invention there is provided a method for selecting a patient for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, wherein said patient is suffering from cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD4+ cells in a blood sample previously isolated from said patient are CD205+; and administering to the patient a therapeutically effective amount of an anti-CD205 antibody, or antigen-binding fragment thereof; is provided.

[0114] According to a further aspect of the invention there is provided an in vitro method for selecting a patient for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, said method comprising the steps of: a. determining the percentage of CD4+ cells in a blood sample previously isolated from said patient that are CD205+ cells; and b. selecting the patient for treatment with an antibody or antigen-binding fragment thereof that binds CD205 if at least 20% of the CD4+ cells in the blood sample are CD205+; is provided.

[0115] In one embodiment, the method for selecting a patient further comprises treating said patient with said antibody, or antigen-binding fragment thereof, that binds to CD205.

[0116] According to another aspect of the invention, there is provided a method for determining the efficacy of an antibody or antigen-binding fragment thereof that binds CD205 in treating cancer in a patient, said method comprising the steps of: a. obtaining a blood sample from the subject; b. identifying whether at least 20% of the CD4+ cells in the blood sample are CD205+; is provided.

[0117] In one embodiment, the method for determining efficacy further comprises administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds CD205 if at least 20% of the CD4+ cells in the blood sample are CD205+.

[0118] In further embodiments, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD4+ cells are CD205+.

[0119] According to a further aspect of the invention there is provided a method for selecting a patient suitable for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, wherein said patient is suffering from cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD8+ and CD4+ cells in a blood sample previously isolated from said patient are CD205+; and administering to the patient a therapeutically effective amount of an anti-CD205 antibody, or antigen-binding fragment thereof; is provided.

[0120] According to yet a further aspect of the invention there is provided an in vitro method for selecting a patient for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, said method comprising the steps of: a. determining the percentage of CD8+ and CD4+ cells in a blood sample previously isolated from said patient that are CD205+ cells; and b. selecting the patient for treatment with an antibody or antigen-binding fragment thereof that binds CD205 if at least 20% of the CD8+ and CD4+ cells in the blood sample are CD205+; is provided.

[0121] In one embodiment, the method for selecting a patient further comprises treating said patient with said antibody, or antigen-binding fragment thereof, that binds to CD205.

[0122] According to another aspect of the invention, there is provided a method for determining the efficacy of an antibody or antigen-binding fragment thereof that binds CD205 in treating cancer in a patient, said method comprising the steps of: obtaining a blood sample from the patient; identifying whether at least 20% of the CD8+ and CD4+ cells in the blood sample are CD205+; is provided.

[0123] In one embodiment, the method for determining efficacy further comprises administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds CD205 if at least 20% of the CD8+ and CD4+ cells in the blood sample are CD205+.

[0124] In further embodiments, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD8+ and CD4+ cells are CD205+.

[0125] According to a further aspect of the present invention there is provided a method for treating or preventing cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD8+ cells in a blood sample previously isolated from said patient are CD205+; and administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds to CD205; is provided.

[0126] Preferably, wherein at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD8+ cells are CD205+.

[0127] According to a further aspect of the present invention there is provided a method for treating or preventing cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD4+ cells in a blood sample previously isolated from said patient are CD205+; and administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds to CD205; is provided.

[0128] Preferably, wherein at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD4+ cells are CD205+.

[0129] According to a further aspect of the present invention there is provided a method for treating or preventing cancer, said method comprising the steps of: Identifying a patient, wherein at least 20% of CD8+ cells and CD4+ cells in a blood sample previously isolated from said patient are CD205+; and administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds to CD205; is provided.

[0130] Preferably, wherein at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the patient's CD8+ and CD4+ cells are CD205+.

[0131] According to a further aspect, a method of treatment comprising the steps of: (a) calculating the percentage of CD4+ and / or CD8+ cells in a blood sample previously isolated from a patient diagnosed with cancer that are CD205+ cells to identify said patient as having a responder phenotype; and (b) administering to said patient having a responder phenotype a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds CD205; is provided.

[0132] As used in this application, the term responder phenotype is defined as a patient in which at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% of the CD4+ and / or CD8+ cells in a blood sample previously isolated from said patient are CD205+ positive.

[0133] In one embodiment, the antibody or antigen-binding portion thereof that binds to CD205 further comprises a covalently-linked moiety. Preferably, the moiety is a drug. More preferably, the drug is selected from the group consisting of maytansinoids, dolastatins, hemiasterlins, auristatins, trichothecenes, calicheamicins, duocarmycins, bacterial immunotoxins, pyranoindoidinoquinolines, camptothecins, anthracyclines, antheamicins, thienoindoles, amatoxins, CC1065, or taxol, and derivatives thereof.

[0134] In a preferred embodiment, the drug is a maytansinoid selected from the group consisting of DM4 and DM1, preferably DM4.

[0135] In some embodiments, the method comprises the further step of then administering to the patient a checkpoint inhibitor.

[0136] In certain embodiments, the checkpoint inhibitor is PD1, PD-L1, PD-L2, CTLA-4, ICOS, TIGIT, CD28, TMIGD2, CD137, CD137L, CD27, OX40, OX40L, LAG3, VISTA, GITR, DNAM-1, CD96, 2B4, TIM-3, CEACAM, CRTAM, SLAMF6, Galectin-9, CD48, CD155, GITRL, CD40, CD40L, CD70, HVEM, B7-H7, B7-H3, B7-H4, ICOSL, CD80, CD86, BTLA, CD160, LIGHT, adenosine A2a receptor, SIRP alpha, DC-SIGN, CD200R, DR3, TL1A, CD200, The antibody targets a checkpoint protein selected from the group including BTN2A1, CD47, IDO, and TDO.

[0137] Preferably, the checkpoint inhibitor is PD1 or PD-L1, more preferably PD1.

[0138] In one embodiment, the PD1 / PD-L1 inhibitor is an antibody.

[0139] In some embodiments, the anti-PD-1 antibody is nivolumab (MDX-1 106, Opdivo; Bristol-Myers Squibb), pembrolizumab (MK-3475, Keytruda, lambrolizumab, BMS-936558; Merck), cemiplimab (REGN-2810, Libtayo; Regeneron), dostallimab (TSR-042, Tesaro, Inc.), EH12.2H7 (ENUM-388D4, BioLegend, catalog number 329902), Balstilimab (Agenus Inc.).

[0140] In further embodiments, the anti-PD-L1 antibody is avelumab (BAVENCIO; EMD Serono, Pfizer), durvalumab (IMFINZI, AstraZeneca), BMS-936559, atezolizumab (TECENTRIQ, Genentech).

[0141] In various embodiments, the checkpoint inhibitor is administered between 7 days and 12 weeks, preferably between 7 days and 10 weeks, or 7 days and 8 weeks, or 7 days and 6 weeks, or 7 days and 4 weeks, or 7 days and 21 days, or 10 days and 19 days, or 12 days and 16 days, or 14 days and 16 days, or 19 days and 28 days, more preferably between 20 days and 25 days, and most preferably between 21 days and 24 days, after administration of an antibody, or antigen-binding portion thereof, that binds CD205.

[0142] Preferably, said patient has not previously been treatable with a checkpoint inhibitor.

[0143] In a further aspect of the invention, there is provided a method for treating or preventing cancer, said patient in need thereof, comprising the step of administering to said patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof which modulates a population of CD205+ immunoregulatory cells, and a therapeutically effective amount of a composition comprising a cancer vaccine; is provided.

[0144] In a further aspect of the invention, there is provided a method for enhancing the effectiveness of a cancer vaccine in a patient, the method comprising administering to the patient (a) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that modulates a population of CD205+ immunoregulatory cells, and (b) a composition comprising the cancer vaccine.

[0145] It will be apparent to one of skill in the art that the antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells and the composition comprising the cancer vaccine may be administered simultaneously, separately, or sequentially.

[0146] Those skilled in the art will appreciate that administration of an antibody or antigen-binding fragment thereof that modulates a population of CD205+ immunoregulatory cells, as described herein, may increase the number of both pDCs and mDCs present in the patient's blood, and may also increase the number of T-cells. Those skilled in the art will further appreciate that this increase may lead to improved responsiveness to a cancer vaccine, due to an increase in the number of dendritic cells that present the antigen encoded by the cancer vaccine, and an increase in the number of T-cells that can be activated by the presented antigen.

[0147] In a further aspect of the invention there is provided a method for treating or preventing cancer, comprising administering to a patient in need thereof a therapeutically effective amount of an antibody or antigen-binding fragment thereof which modulates a population of CD205+ immunoregulatory cells, and a therapeutically effective amount of a composition comprising a bispecific antibody; is provided.

[0148] In one embodiment, the bispecific antibody is a bispecific T-cell engager (BiTE). Preferably, the bispecific antibody comprises a first binding domain that binds to CD3. More preferably, the bispecific antibody comprises a second binding domain that binds to a tumor-specific antigen.

[0149] It will be apparent to one skilled in the art that the antibody or antigen-binding fragment thereof that regulates the population of CD205+ immunoregulatory cells and the composition comprising the bispecific antibody may be administered simultaneously, separately or sequentially.

[0150] In a further aspect, a method for enhancing the effect of a bispecific (preferably, BiTE) antibody in a patient, wherein the patient has been identified as one in need thereof, comprising administering to the patient (a) a therapeutically effective amount of an antibody or antigen-binding fragment thereof that modulates a population of CD205+ immunoregulatory cells, and (b) a composition comprising a bispecific antibody.

[0151] One skilled in the art will appreciate that following administration of an antibody or antigen-binding fragment thereof that modulates the population of CD205+ immunoregulatory cells, the number of T-cells will increase, and therefore the number of cells that can be activated and brought into close proximity to the target cells by the bispecific antibody (preferably BiTE), thus increasing the effectiveness in treating cancer.

[0152] The skilled artisan will appreciate that the bispecific antibody may be any suitable bispecific antibody, preferably a BiTE, including, but not limited to, bispecific antibodies that bind to CD19 and CD3, Epcam and CD3, DLL3 and CD3, or B7H6 and CD3.

[0153] The present invention also provides a method for treating cancer in a subject, said method comprising: a. obtaining a tumor sample from the subject; b. immunohistochemically staining the tumor sample to identify whether at least 50% of the tumor cells in the tumor sample express DCE205 at a level of at least 2+; c. administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds CD205 if at least 50% of the tumor cells in the tumor sample express CD205 at a level of at least 2+; Also provided.

[0154] In a further aspect, the invention provides a method for treating cancer in a human patient, comprising: identifying a patient having a tumor, wherein at least 50% of the tumor cells express CD205 at a level of 2+ as measured by immunohistochemistry (IHC); and administering to the patient a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds CD205. to provide.

[0155] According to a further aspect, there is provided a method of selecting a patient suitable for anti-CD205 antibody therapy, said method comprising: identifying a patient having a tumor in which at least 50% of the tumors have CD205 expression at a level of 2+ as measured by immunohistochemistry (IHC); and instructing a health care provider to administer an anti-CD205 antibody or antigen-binding fragment thereof to said patient. is provided.

[0156] According to a further aspect of the invention there is provided an in vitro method for selecting a cancer patient for treatment with an antibody or antigen-binding fragment thereof that binds to CD205, said method comprising the steps of: determining the expression level of CD205 in a tumor sample isolated from said patient; and selecting the patient for treatment with the antibody or antigen-binding fragment thereof that binds CD205 if the tumor sample shows an expression level of 2+ in at least 50% of the tumor cells as measured by immunohistochemistry (IHC); is provided.

[0157] In one embodiment, the in vitro method further comprises the step of treating the patient with the antibody, or antigen-binding fragment thereof, that binds CD205.

[0158] In a further aspect of the invention there is provided a method for determining the efficacy of an antibody or antigen-binding fragment thereof that binds CD205 in treating cancer in a subject, said method comprising: obtaining a tumor sample from the subject; immunohistochemically staining the tumor sample to identify whether at least 50% of the tumor cells in the tumor sample express DCE205 at a level of at least 2+; is provided.

[0159] In one embodiment, the method further includes administering to the subject a therapeutically effective amount of an antibody or antigen-binding fragment thereof that binds to CD205 if at least 50% of the tumor cells in the tumor sample express CD205 at a level of at least 2+.

[0160] In further embodiments, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of the tumor cells in the tumor sample express DEC 205 at a level of at least 2+ as measured by IHC.

[0161] It will be readily apparent that IHC may be performed using any suitable protocol and any suitable antibody that specifically binds to CD205 on tumor samples. In one embodiment, the antibody is an anti-CD205 antibody from Leica (catalog number: NCL-L-CD205).

[0162] In one embodiment, the tumor sample is in the form of a formalin fixed paraffin embedded (FFPE) sample. In another embodiment, the sample is a fresh frozen tumor sample. Also within the scope of the present invention is a kit comprising the pharmaceutical combination of the present invention and, optionally, instructions for use. The kit may further comprise at least one additional reagent or one or more additional antibodies.

[0163] Other features and advantages of the invention will become apparent from the following detailed description and claims. [Brief description of the drawings]

[0164] [Figure 1] Figure 1 shows the sequence of the CD205_A1 antibody heavy chain variable region (SEQ ID NO: 1). The CDR regions of the CD205_A1 antibody heavy chain are underlined. [Diagram 2] Figure 2 shows the sequence of the CD205_A1 antibody light chain variable region (SEQ ID NO:2). The CDR regions of the CD205_A1 antibody light chain are underlined. [Diagram 3] Figure 3 shows, in the left panel, the changes in the number of CD8+ T-cells in blood samples taken from gastric cancer patients on days 1, 8, 15, and 21 after treatment with 2.5 mg / kg anti-CD205-DM4 ADC, and in the right panel, the changes in the number of CD4+ T-cells over time. [Figure 4] Figure 4 shows in the left panel the change in the percentage of the total T-cell population composed of CD4+ (upper panel) and CD8+ (lower panel) T-cells over a 21-day time series following treatment with 2.5 mg / kg anti-CD205-DM4 ADC. The right panel shows the change in the percentage of CD4+ and CD8+ T cells that are PD1+ over a 21-day time series. [Diagram 5]Figure 5 shows in the left panel the evolution over time of the number of CD8+ T-cells present in the patient's blood that are also PD1+, and in the right panel the evolution over time of the number of CD4+ T-cells present in the patient's blood that are also PD1+. [Figure 6] FIG. 6 shows the change in the number of CD8+ CD205+ cells over a 21-day time course following treatment with 2.5 mg / kg anti-CD205-DM4 ADC. [Figure 7] FIG. 7 shows the change in CD4+ CD205+ cell numbers over a 21-day time course following treatment with 2.5 mg / kg anti-CD205-DM4 ADC. [Figure 8] Figure 8 shows, in the left panel, the numbers of mDC and pDC in blood samples taken from a gastric cancer patient on days 1, 8, 15, and 21 after treatment with 2.5 mg / kg anti-CD205-DM4 ADC, and the right panel shows the change in the numbers of CD205+ mDC and pDC in the patient's blood over a 21-day time period. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0165] Detailed Description of the Invention The present disclosure relates to methods for increasing immune responses in patients suffering from cancer and for increasing the efficacy of immune checkpoint inhibitors. It also discloses a combination pharmaceutical comprising an anti-CD205 antibody and an immune checkpoint inhibitor, wherein the combination pharmaceutical is in the form of a combined preparation for separate or sequential use.

[0166] CD205 protein CD205 is thought to function as an endocytic receptor that targets trapped antigens from the extracellular space to specialized antigen-compartments, resulting in a decrease in B-lymphocyte proliferation.

[0167] According to UNIPROT, CD205 is expressed in spleen, thymus, colon and peripheral blood lymphocytes. It has been detected in myeloid and B-lymphoid cell lines. The isoforms OGTA076b and OGTA076c are expressed in malignant Hodgkin's lymphoma cells, called Hodgkin and Reed-Sternberg (HRS) cells. CD205 functions as an endocytic receptor that targets trapped antigens from the extracellular space to specialized antigen-compartments. It reduces the proliferation of B-lymphocytes.

[0168] Expression of CD205 has been observed in gastric, pancreatic, bladder, ovarian, breast (e.g. Her2-ve and triple negative), colorectal, renal, endometrial, gastroesophageal junction, esophageal, skin, thyroid and lung (non-small-cell) cancers, as well as in multiple myeloma, and many different subtypes of lymphomas (including DLBCL) and leukemias.

[0169] Anti-CD205 antibodies or antigen-binding portions thereof for use in the methods or combinations of the invention may, in certain cases, cross-react with CD205 from species other than human. For example, to facilitate clinical trials, the anti-CD205 antibodies may cross-react with mouse or primate CD205. Alternatively, in certain embodiments, the antibodies may be entirely specific for human CD205 and may not exhibit species or other types of non-human cross-reactivity.

[0170] PD-L1 protein According to UNIPROT, PD-L1 is a type I membrane protein. The protein consists of an extracellular domain, between amino acids 19-238, consisting of one Ig-like V-type (immunoglobulin-like) domain, one Ig-like C2-type (immunoglobulin-like) domain; it further consists of one transmembrane region and one cytoplasmic region.

[0171] In some embodiments, the antibody for use in the methods or combinations of the invention binds to human PD-L1.

[0172] Antibodies for use in accordance with embodiments of the invention may, in some cases, cross-react with PD-L1 proteins from species other than human. For example, to facilitate preclinical and toxicity testing, antibodies of the invention may cross-react with mouse or primate PD-L1 proteins. Alternatively, in certain embodiments, antibodies for use in the methods of the invention may be specific for human PD-L1 protein and may not exhibit non-human species or other types of cross-reactivity.

[0173] PD1 protein According to UNIPROT, PD1 is an inhibitory receptor on antigen-activated T-cells that plays an important role in the induction and maintenance of immune tolerance to self. PD1 transmits inhibitory signals upon binding to its ligands CD274 / PDL1 and CD273 / PDLG2.

[0174] PD1-mediated inhibitory pathways are exploited by tumors to attenuate anti-tumor immunity and avoid destruction by the immune system, thereby promoting tumor survival. Interaction with CD274 / PDL1 inhibits cytotoxic T lymphocyte (CTL) effector function. Blocking PD1-mediated pathways reverses the exhausted T-cell phenotype and normalizes anti-tumor responses. This provides a rationale for cancer immunotherapy.

[0175] In some embodiments, an antibody for use in the methods or combinations of the invention binds to human PD1.

[0176] Antibodies for use according to embodiments of the invention may, in certain cases, cross-react with PD1 proteins from species other than human. For example, to facilitate pre-clinical and toxicity testing, antibodies of the invention may cross-react with murine or primate PD1 proteins. Alternatively, in certain embodiments, antibodies for use in methods of the invention may be specific for human PD1 proteins and may not exhibit non-human species or other types of cross-reactivity.

[0177] antibody Antibodies that find use in the methods of the invention may take many formats, e.g., conventional antibodies, as well as antibody derivatives, fragments, and mimetics, as described herein. In one embodiment, an antibody structure is provided that comprises the six CDR sets defined in the present application (with minor amino acid changes as described below).

[0178] "Antibody" as used herein, as will be understood by one of skill in the art, encompasses a wide variety of structures that, in some embodiments, comprise at least the six CDR sets defined herein; such as, but not limited to, conventional antibodies (including both monoclonal and polyclonal antibodies), humanized and / or chimeric antibodies, antibody fragments, modified antibodies (e.g., having amino acid modifications as outlined below), multispecific antibodies (e.g., bispecific antibodies, etc.), and other analogs known in the art.

[0179] A conventional antibody structural unit typically includes a tetramer. Each tetramer has two identical pairs of polypeptide chains, each pair having one "light" chain (typically having a molecular weight of about 25 kDa) and one "heavy" chain (typically having a molecular weight of about 50-70 kDa). The amino-terminal portion of each chain contains a variable region of about 100 to 110 or more amino acids that is primarily responsible for antigen recognition. In the variable region, three loops are assembled for each of the heavy and light chain V domains to form the antigen-binding site. Each of the loops is called a complementarity-determining region (hereinafter referred to as "CDR"), in which the amino acid sequence variation is most pronounced. "Variable" refers to the fact that certain segments of the variable region have large sequence differences between antibodies. The variations within the variable region are not evenly distributed. Instead, V regions consist of relatively invariant stretches of 15-30 amino acids, called framework regions (FR), separated by shorter regions that are highly variable, called "hypervariable regions", each 9-15 amino acids long or longer.

[0180] Each of VH and VL is composed of three hypervariable regions ("complementarity determining regions", "CDRs") and four FRs, arranged from the amino terminus to the carboxy terminus in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.

[0181] The hypervariable region generally includes amino acid residues from about amino acid residues 24-34 (LCDR1; "L" indicates light chain), 50-56 (LCDR2), and 89-97 (LCDR3) in the light chain variable region, and about amino acid residues 31-35B (HCDR1; "H" indicates heavy chain), 50-65 (HCDR2), and 95-102 (HCDR3) in the heavy chain variable region; Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991) and / or those residues form hypervariable loops (e.g., residues 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3) in the light chain variable region and 26-32 (HCDR1), 53-55 (HCDR2) and 96-101 (HCDR3) in the heavy chain variable region); Chothia and Lesk (1987) J. Mol. Biol. 196:901-917. Particular CDRs of the invention are described below.

[0182] Throughout this specification, the Kabat numbering system is generally used when referring to residues in the variable domain (approximately residues 1-107 in the light chain variable region and residues 1-113 in the heavy chain variable region) (e.g., Kabat et al., supra (1991)).

[0183] The CDRs contribute to the formation of the antigen-binding site, more specifically the epitope-binding site of an antibody. The term "epitope" or "antigenic determinant" refers to a site on an antigen to which an immunoglobulin or antibody specifically binds. Epitopes can be formed both from contiguous amino acids or from non-contiguous amino acids juxtaposed by tertiary folding of a protein.

[0184] In some embodiments, the antibodies for use in the methods of the invention are full length. By "full length antibody" is meant the structure that constitutes the native biological form of an antibody, including variable and constant regions, including one or more modifications as outlined in this application.

[0185] Alternatively, antibodies for use in the methods of the invention can be in a variety of configurations, including, but not limited to, antibody fragments, monoclonal antibodies, bispecific antibodies, minibodies, domain antibodies, synthetic antibodies (sometimes referred to in the present application as "antibody mimetics"), chimeric antibodies, humanized antibodies, antibody fusions (sometimes referred to as "antibody conjugates"), chimeric antigen receptors (CARs), and fragments of each. Configurations that rely on the use of a set of CDRs are included within the definition of "antibody."

[0186] In one embodiment, an antibody for use in the methods of the invention is an antibody fragment. Specific antibody fragments include, but are not limited to, (i) a Fab fragment consisting of the VL, VH, CL, and CH1 domains, (ii) an Fd fragment consisting of the VH and CH1 domains, (iii) an Fv fragment consisting of the VL and VH domains of a single antibody, (iv) a dAb fragment consisting of a single variable region (Ward et al., 1989, Nature 341:544-546, which is incorporated by reference in its entirety), (v) an isolated CDR region, (vi) an F(ab')2 fragment, a bivalent fragment comprising two linked Fab fragments, and (vii) a single-chain Fv molecule (scFv), in which the VH and VL domains are linked by a peptide linker that allows the two domains to associate to form an antigen-binding site (Bird et al., 1988, Science 242:423-426, Huston et al., 1988, Proc. Natl. Acad. Sci. USA 85:5879-5883, which are incorporated by reference in their entireties), (viii) bispecific single chain Fvs (WO 03 / 11161, which is incorporated by reference herein), and (ix) "diabodies" or "triabodies", multivalent or multispecific fragments constructed by genetic fusion (Tomlinson et. al., 2000, Methods Enzymol. 326:461-479; WO94 / 13804; Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-6448, all of which are incorporated by reference in their entireties).

[0187] Chimeric and Humanized Antibodies In some embodiments, the antibody may be a mixture of different species origins, e.g., a chimeric antibody and / or a humanized antibody, i.e., the present invention may use a set of CDRs with framework and constant regions other than those specifically described by sequence in this application.

[0188] In one embodiment, antibodies for use in the methods of the invention may be multispecific antibodies, particularly bispecific antibodies, also sometimes referred to as "diabodies." These are antibodies that bind to two (or more) different antigens, or different epitopes on the same antigen. Diabodies can be produced in a variety of ways known in the art (Holliger and Winter, 1993, Current Opinion Biotechnol. 4:446-449, incorporated by reference in its entirety), for example, prepared chemically or from hybrid hybridomas.

[0189] In one embodiment, the antibody for use in the methods of the invention is a minibody. Minibodies are minimized antibody-like proteins that contain an scFv linked to a CH3 domain. Hu et al., 1996, Cancer Res. 56:3055-3061, incorporated by reference in its entirety. In some cases, the scFv may be linked to the Fc region and may include part or the entire hinge region. Note that minibodies are included within the definition of "antibody" despite the fact that they do not have a complete set of CDRs.

[0190] The antibodies disclosed for use in the methods described in this application may be isolated or recombinant. "Isolated," as used to describe various polypeptides disclosed herein, refers to a polypeptide that has been identified and separated and / or recovered from a cell expressing the polypeptide or from a cell culture. Thus, an isolated antibody is intended to refer to an antibody that is substantially free of other antibodies with various antigen specificities (e.g., an isolated antibody that specifically binds to CD205 is substantially free of antibodies that specifically bind to antigens other than CD205). Thus, an "isolated" antibody is an antibody that is found in a form not normally found in nature (e.g., non-naturally occurring). An isolated antibody as defined in this application may, in one embodiment, include at least one amino acid that is not present in a "naturally" occurring antibody. This amino acid may be introduced by addition or substitution. It will be understood that the introduced amino acid may be a naturally occurring or non-naturally occurring amino acid. In some embodiments, the antibody of the invention is a recombinant protein, an isolated protein, or a substantially pure protein. An "isolated" protein is free from at least some of the materials with which it is normally associated in its natural state, e.g., it comprises at least about 5% by weight, or at least about 50% by weight, of the total protein in a given sample. It is understood that the isolated protein may comprise from 5 to 99.9% by weight of the total protein content, depending on the circumstances. For example, the protein may be produced at a significantly higher concentration by using an inducible promoter or a high expression promoter, so that the protein is produced at an increased concentration level. In the case of recombinant proteins, the definition includes the production of antibodies in a wide variety of organisms and / or host cells known in the art that do not naturally produce them. Usually, isolated polypeptides are prepared by at least one purification step. An "isolated antibody" refers to an antibody that is substantially free of other antibodies with different antigen specificities.For example, an isolated antibody that specifically binds CD205 is substantially free of antibodies that specifically bind antigens other than CD205.

[0191] Isolated monoclonal antibodies with different specificities may be combined into well-defined compositions, and thus, for example, the antibodies of the invention may be optionally and individually included or excluded from a formulation, as discussed further below.

[0192] Anti-CD205 antibodies for use in the present invention specifically bind to CD205 (e.g., SEQ ID NO: 11). "Specific binding" or "specifically binds" or "specific" for a particular antigen or epitope means binding that is measurable as distinct from non-specific interactions. Specific binding can be measured, for example, by determining binding of a molecule compared to binding of a control molecule, which is generally a molecule of similar structure that has no binding activity. For example, specific binding may be determined by competition with a control molecule that is similar to the target.

[0193] For example, an antibody having a KD for an antigen or epitope of at least about 10-4 M, at least about 10-5 M, at least about 10-6 M, at least about 10-7 M, at least about 10-8 M, at least about 10-9 M, or at least about 10-10 M, at least about 10-11 M, at least about 10-12 M, or stronger, indicates specific binding to a particular antigen or epitope, where KD refers to the off-rate of a particular antibody-antigen interaction. Typically, an antibody that specifically binds to an antigen has a KD that is 20-fold, 50-fold, 100-fold, 500-fold, 1,000-fold, 5,000-fold, 10,000-fold, or more greater for a control molecule compared to the antigen or epitope. However, in the present invention, when administering an ADC of an antibody of CD205 according to the present invention, what is important is that the KD is sufficient to allow internalization and cell death without significant side effects.

[0194] Additionally, an antibody having a K A or K A for an antigen or epitope that is at least 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more greater for that epitope compared to a control, indicates specific binding for a particular antigen or epitope, where K A or K A refers to the off-rate of a particular antibody-antigen interaction.

[0195] Standard assays for evaluating the binding ability of antibodies to CD205 may be performed at the protein or cellular level and are known in the art, such as, for example, ELISA, Western blot, RIA, BIAcore® assay, and flow cytometry analysis. Suitable assays are described in detail in the present Examples. The binding kinetics (e.g., binding affinity) of the antibodies may also be evaluated by standard assays known in the art, such as Biacore® system analysis. To evaluate binding to Raji or Daudi B cell tumor cells, Raji (ATCC Accession No. CCL-86) or Daudi (ATCC Accession No. CCL-213) cells can be obtained from publicly available sources, such as the American Type Culture Collection, and used in standard assays, such as flow cytometry analysis.

[0196] CD205 antibody CD205 antibodies for use in the methods of the invention bind to CD205 (SEQ ID NO: 11) and are internalized when contacted with cells expressing CD205 on the cell surface. These antibodies are referred to in this application as either "anti-CD205" antibodies, or, for ease of description, "CD205 antibodies." Both terms are used interchangeably in this application.

[0197] The CD205 antibodies for use in the methods of the invention are internalized upon contact with cells expressing CD205 on their surface, particularly tumor cells. That is, the CD205 antibodies as defined in the present application, including drug conjugates, are internalized by tumor cells, resulting in the release of the drug and subsequent cell death, allowing the treatment of cancers that express CD205. Internalization in this context can be measured in several ways. In one embodiment, the CD205 antibodies are contacted with cells, such as the cell lines outlined in the present application, using a standard assay such as MAbZap. It will be clear to one skilled in the art that the MAbZap assay represents the expected behavior of antibody drug conjugates (ADCs). In the latter case, the ADC is internalized and therefore the drug is internalized in the cell. A toxic drug will have the ability to damage the cell, i.e., to damage the targeted cancer cells. Data from the MabZap assay are readily recognized by those of skill in the art as being representative of ADC assays (Kohls, M and Lappi, D.,

[2000] Biotechniques, vol. 28, no. 1, 162-165).

[0198] In one embodiment, an anti-CD205 antibody for use in the methods of the invention comprises the heavy and light chain complementarity determining regions (CDRs) or variable regions (VRs) of a particular antibody described in the present application (e.g., referred to in the present application as "CD205_A1"). Thus, in one embodiment, an antibody for use in the methods of the invention comprises the CDR1, CDR2, and CDR3 domains of the heavy chain variable (VH) region of antibody CD205_A1 having the sequence set forth in SEQ ID NO:1, and the CDR1, CDR2, and CDR3 domains of the light chain variable (VL) region of antibody CD205_A1 having the sequence set forth in SEQ ID NO:2.

[0199] In another embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a heavy chain variable region comprising a first vhCDR comprising SEQ ID NO:5; a second vhCDR comprising SEQ ID NO:6; and a third vhCDR comprising SEQ ID NO:7; and a light chain variable region comprising a first vlCDR comprising SEQ ID NO:8; a second vlCDR comprising SEQ ID NO:9; and a third vlCDR comprising SEQ ID NO:10.

[0200] In another embodiment, an anti-CD205 antibody for use in the methods of the invention binds human CD205 and comprises a heavy chain variable region comprising an amino acid sequence comprising SEQ ID NO:1, and conservative sequence modifications thereof. An antibody for use in the methods of the invention may further comprise a light chain variable region comprising an amino acid sequence comprising SEQ ID NO:2, and conservative sequence modifications thereof.

[0201] In a further embodiment, an anti-CD205 antibody for use in the methods of the invention binds to human CD205 and comprises a heavy chain variable region and a light chain variable region comprising one of the sequence combinations set forth in Table 1 below: [Table 1]

[0202] In a further embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a heavy chain variable region and a light chain variable region that bind human CD205 and that comprise the amino acid sequences set forth in SEQ ID NO: 1 and / or 2, respectively, and conservative sequence modifications thereof. As used in this application, the term conservative sequence modification refers, for example, to the replacement of an amino acid with an amino acid having similar characteristics. It is common knowledge for those skilled in the art what substitutions are considered conservative. Other modifications that are considered to be conservative sequence modifications include, for example, glycosylation.

[0203] Optionally, one or more of SEQ ID NOs:5-10 independently include one, two, three, four, or five conservative amino acid substitutions; optionally, one or more of SEQ ID NOs:5-10 independently include one or two conservative amino acid substitutions.

[0204] Preferably, the term "conservative sequence modifications" is intended to include amino acid modifications that do not significantly affect or change the binding characteristics of an antibody containing said amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. Modifications may be introduced into an antibody of the present invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are those in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within the CDR regions of an antibody of the invention may be replaced with another amino acid residue from the same side chain family, and the altered antibodies may be tested for retained function using other functional assays described herein.

[0205] In one embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a heavy chain variable region comprising SEQ ID NO: 1, or a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1. In another embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a light chain variable region comprising SEQ ID NO: 2, or a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 2. In another embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a heavy chain framework region comprising an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a framework of the heavy chain variable region of SEQ ID NO: 1, including SEQ ID NOs: 12, 13, 14 and 15. In another embodiment, an anti-CD205 antibody for use in the methods of the invention comprises a light chain framework region comprising an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identical to a framework of the light chain variable region of SEQ ID NO: 2, including SEQ ID NOs: 16, 17, 18 and 19.

[0206] In one embodiment, an anti-CD205 antibody for use in the methods of the invention is referred to in the present application as a "CD205_A1 antibody" comprising the following CDRs, as well as variants that include a limited number of amino acid variants. [Table 2]

[0207] The present application also discloses variable heavy and light chains, as well as full-length heavy and light chains (including, for example, constant regions) comprising the CDR set of the present invention. As will be understood by those skilled in the art, the CDR set of the anti-CD205 antibody may be incorporated into a murine, humanized, or human constant region (including framework region). Thus, the present disclosure provides variable heavy and variable light chains that are at least about 90%-99% identical to the sequence numbers (SEQ IDs) disclosed in the present application, and it is understood that 90, 91, 92, 93, 94, 95, 96, 97, 98, and 99% are all useful in the present invention.

[0208] In one embodiment, an antibody for use in the methods of the invention specifically binds to human CD205 comprising SEQ ID NO: 11. Preferably, an anti-CD205 antibody for use in the methods of the invention binds with high affinity to human CD205.

[0209] Antibody Modification The invention further provides variant antibodies, sometimes referred to as "antibody derivatives" or "antibody analogs", for use in the methods of the invention. Thus, there are many modifications that can be made to the antibodies of the invention, including, but not limited to, amino acid modifications in the CDRs (affinity maturation), amino acid modifications in the framework region, amino acid modifications in the Fc region, glycosylation variants, and other types of covalent modifications (e.g., for attachment of drug conjugates).

[0210] By "variant" in the present application is meant a polypeptide sequence that differs from that of a parent polypeptide by at least one amino acid modification. In this case, the parent polypeptide is either the full-length variable heavy or light chain as listed in SEQ ID NOs: 1 or 2, or the CDR or framework regions of the heavy and light chains as listed in SEQ ID NOs: 5-10 and 12-19, respectively. The amino acid modifications may include substitutions, insertions and deletions, with the former being preferred in many cases. It is understood that the amino acid substitutions may be conservative or non-conservative, with conservative substitutions being preferred. Furthermore, the substitutions may be with either naturally occurring or non-naturally occurring amino acids.

[0211] As used herein, an "amino acid substitution" or "substitution" refers to the replacement of an amino acid at a particular position in a parent polypeptide sequence with another amino acid, which may be a naturally occurring or non-naturally occurring amino acid. For example, the substitution S100A refers to a variant polypeptide in which the serine at position 100 is replaced with an alanine. As used herein, an "amino acid insertion" or "insertion" refers to the addition of an amino acid at a particular position in a parent polypeptide sequence. As used herein, an "amino acid deletion" or "deletion" refers to the removal of an amino acid at a particular position in a parent polypeptide sequence.

[0212] "Parent polypeptide", "parent protein", "precursor polypeptide" or "precursor protein" as used herein refers to an unmodified polypeptide which is subsequently modified to produce a variant. Generally, the parent polypeptide in this application is LY75_A1. Thus, a "parent antibody" as used herein refers to an antibody which has been modified to produce a variant antibody.

[0213] As used herein, "wild-type" or "WT" or "native" refers to an amino acid sequence or nucleotide sequence found in nature, including allelic variations. A WT protein, polypeptide, antibody, immunoglobulin, IgG, etc., has an amino acid sequence or nucleotide sequence that has not been intentionally modified.

[0214] By "variant Fc region" herein is meant an Fc sequence that differs from a wild-type Fc sequence by virtue of at least one amino acid modification. An Fc variant may refer to the Fc polypeptide itself, an Fc variant polypeptide, or a composition comprising the amino acid sequence.

[0215] In some cases, the amino acid modification in the CDR is referred to as "affinity maturation". An "affinity matured" antibody is an antibody that has one or more modifications in one or more CDRs, which results in an improved affinity of the antibody for the antigen compared to a parent antibody that does not have those modifications. In some cases, although rare, it may be desirable to decrease the affinity of an antibody for its antigen, but this is generally not preferred.

[0216] Alternatively, amino acid modifications may be made in one or more of the CDRs of an antibody of the invention that are "silent", e.g., do not significantly alter the affinity of the antibody for the antigen. These may be made for a number of reasons, such as to optimize expression (as may be done for nucleic acids encoding an antibody of the invention).

[0217] Thus, variant CDRs and antibodies are included within the definition of the CDRs and antibodies of the invention; i.e., the antibodies of the invention may comprise amino acid modifications in one or more CDRs of LY75_A1. In addition, as outlined below, amino acid modifications may also be made independently and optionally in any region outside of the CDRs, including the framework and constant regions as described in this application.

[0218] In some embodiments, the anti-LY75 antibody is comprised of a variant Fc domain. As is known in the art, the Fc region of an antibody interacts with many Fc receptors and ligands, conferring a set of important functional capabilities referred to as effector functions. Furthermore, cysteine ​​modifications are particularly beneficial in antibody-drug conjugate (ADC) applications, as further described below. In some embodiments, the constant region of the antibody may be engineered to contain one or more cysteines that are specifically "thiol-reactive," allowing for more specific and controlled placement of drug moieties. See, e.g., U.S. Patent No. 7,521,541, the entirety of which is incorporated by reference herein.

[0219] Antibody-drug conjugates In some embodiments, the anti-CD205 antibodies or antigen-binding portions thereof for use in the methods of the invention disclosed herein are conjugated with a drug to form an antibody-drug conjugate (ADC). Generally, ADCs are used in oncology applications, where the use of antibody-drug conjugates to locally deliver cytotoxic or cytostatic drugs allows for targeted delivery of the drug moiety to the tumor, thereby allowing for higher efficacy, lower toxicity, etc. Overviews of this technology are provided in Ducry et al., Bioconjugate Chem., 21:5-13 (2010), Carter et al., Cancer J. 14(3):154 (2008) and Senter, Current Opin. Chem. Biol. 13:235-244 (2009), all of which are incorporated by reference in their entirety into this application.

[0220] Thus, the present invention provides, inter alia, a combination pharmaceutical comprising an anti-CD205 antibody conjugated to a drug. Generally, conjugation is performed by covalent attachment to the antibody, as further described below, and generally by a linker, often a peptide bond, which may or may not be designed to be susceptible to cleavage by a protease at the target site, as described below. Furthermore, as described above, the linker-drug unit (LU-D) may be linked by attachment to a cysteine ​​in the antibody. As will be appreciated by those skilled in the art, the number of drug moieties per antibody may vary, depending on the conditions of the reaction, and may vary from 1:1 to 10:1 drug:antibody. As will be appreciated by one of ordinary skill in the art, the actual numbers are averages.

[0221] Thus, the anti-CD205 antibody may be conjugated to a drug. As described below, the drug of the ADC may be any number of agents, including, but not limited to, a chemotherapeutic agent, a growth inhibitory agent, a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or a fragment thereof), or a cytotoxic agent such as a radioisotope (i.e., a radioconjugate). In other embodiments, the invention further provides methods of using the ADC.

[0222] In the present invention, drugs for use include cytotoxic drugs, particularly drugs used in cancer therapy. Such drugs generally include DNA damaging drugs, antimetabolites, natural products and their analogs. Exemplary classes of cytotoxic drugs include enzyme inhibitors such as dihydrofolate reductase inhibitors and thymidylate synthase inhibitors, DNA intercalators, DNA cleavage agents, topoisomerase inhibitors, the anthracycline family of drugs, vinca drugs, mitomycins, bleomycins, cytotoxic nucleosides, the pteridine family of drugs, diynenes, podophyllotoxins, dolastatins, maytansinoids, differentiation inducers, and taxol.

[0223] Members of these classes include, for example, taxol, methotrexate, methopterin, dichloromethotrexate, 5-fluorouracil, 6-mercaptopurine, cytosine arabinoside, melphalan, leurosine, leurosideine, actinomycin, daunorubicin, doxorubicin, mitomycin C, mitomycin A, caminomycin, aminopterin, tallysomycin, podophyllotoxin and podophyllotoxin derivatives (e.g., etoposide or etoposide phosphate). etc.), vinblastine, vincristine, vindesine, taxanes (e.g., taxol, taxotere), retinoic acid, butyric acid, N8-acetyl spermidine, camptothecin, calicheamicin, esperamicin, ene-diine, duocarmycin A, duocarmycin SA, calicheamicin, camptothecin, hemiasterlin, maytansinoids (e.g., DM1, etc.), monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), maytansinoid (DM4) and analogs thereof.

[0224] Toxins may be used as antibody-toxin conjugates, and include bacterial toxins such as diphtheria toxin, plant toxins such as ricin, small molecule toxins such as geldanamycin (Mandler et al (2000) J. Nat. Cancer Inst. 92(19):1573-1581; Mandler et al (2000) Bioorganic & Med. Chem. Letters 10:1025-1028; Mandler et al (2002) Bioconjugate Chem. 13:786-791), maytansinoids (EP 1391213; Liu et al., (1996) Proc. Natl. Acad. Sci. USA 93:8618-8623), and calicheamicin (Lode et al (1998) Cancer Res. 58:2928; Hinman et al (1993) Cancer Res. 53:3336-3342), and hemiasterlin (WO2004 / 026293; Zask et al., (2004) J. Med. Chem, 47: 4774-4786). Toxins may exert their cytotoxic and cytostatic effects by mechanisms such as, for example, tubulin binding, DNA binding, or topoisomerase inhibition.

[0225] Conjugates of anti-CD205 antibodies and small molecule toxins, such as maytansinoids, dolastatins, auristatins, trichothecenes, calicheamicins, and CC1065, and derivatives of these toxins, which have toxic activity, may also be used.

[0226] Preferably, the anti-CD205 antibody is conjugated to DM1 or DM4, most preferably DM4.

[0227] Linker Unit Typically, the antibody-drug conjugate compound includes a linker unit between the drug unit and the antibody unit. In some embodiments, the linker is cleavable under intracellular or extracellular conditions, such that cleavage of the linker releases the drug unit from the antibody in the appropriate environment. For example, solid tumors that secrete certain proteases may serve as targets for cleavable linkers; in other embodiments, it is intracellular proteases that are utilized. In yet other embodiments, the linker unit is not cleavable, and the drug is released, for example, by antibody degradation in lysosomes.

[0228] In some embodiments, the linker is cleavable by a cleavage factor present in the intracellular environment (e.g., in a lysosome, or endosome, or caveolae). The linker can be a peptidyl linker that is cleaved by an intracellular peptidase or protease enzyme (such as, but not limited to, a lysosomal protease or an endosomal protease). In some embodiments, the peptidyl linker is at least 2 amino acids in length, or at least 3 amino acids in length, or more.

[0229] Cleavage agents include, but are not limited to, cathepsins B and D, and plasmin, all of which are known to hydrolyze dipeptide drug derivatives, thereby releasing the active drug inside target cells (see, e.g., Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). Peptidyl linkers cleavable by enzymes present in CD205-expressing cells. For example, peptidyl linkers cleavable by cathepsin-B, a thiol-dependent protease highly expressed in cancerous tissues, may be used (e.g., Phe-Leu or Gly-Phe-Leu-Gly linkers). Other examples of such linkers are described, for example, in U.S. Pat. No. 6,214,345, the entirety of which is incorporated herein by reference for all purposes.

[0230] In some embodiments, the peptidyl linker cleavable by an intracellular protease is a Val-Cit linker or a Phe-Lys linker (see, e.g., U.S. Pat. No. 6,214,345, which describes the synthesis of doxorubicin with a val-cit linker).

[0231] In another embodiment, the cleavable linker is pH-sensitive, i.e., sensitive to hydrolysis at certain pH values. Typically, the pH-sensitive linker is hydrolyzable under acidic conditions.

[0232] In yet other embodiments, the linker is cleavable under reducing conditions (eg, a disulfide linker).

[0233] In other embodiments, the linker is a malonic acid linker (Johnson et al., 1995, Anticancer Res. 15:1387-93), a maleimidobenzoyl linker (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1299-1304), or a 3'-N-amide analog (Lau et al., 1995, Bioorg-Med-Chem. 3(10):1305-12).

[0234] In yet other embodiments, the linker unit is not cleavable and the drug is released by antibody degradation. (See U.S. Publication No. 2005 / 0238649, which is incorporated herein by reference in its entirety and for all purposes.)

[0235] In many embodiments, the linker is self-immolative. As used herein, the term "self-immolative spacer" refers to a bifunctional chemical moiety that can covalently link two spaced apart chemical moieties to one another into a stable three-part molecule. It spontaneously separates from the second chemical moiety when its bond to the first moiety is cleaved. See, for example, WO 2007 / 059404A2, WO06 / 110476A2, WO05 / 112919A2, WO2010 / 062171, WO09 / 017394, WO07 / 089149, WO07 / 018431, WO04 / 043493 and WO02 / 083180.

[0236] Often, the linker is substantially insensitive to the extracellular environment. As used herein, in the context of a linker, "substantially insensitive to the extracellular environment" means that no more than about 20%, 15%, 10%, 5%, 3%, or about 1% of the linkers in a sample of the antibody-drug conjugate compound are cleaved when the antibody-drug conjugate compound is present in an extracellular environment (e.g., in plasma).

[0237] In other non-mutually exclusive embodiments, the linker promotes cellular internalization. In certain embodiments, the linker promotes cellular internalization when conjugated to a therapeutic agent (i.e., in the context of a linker-therapeutic agent moiety of an antibody-drug conjugate compound described herein). In yet other embodiments, the linker promotes cellular internalization when conjugated to both an auristatin compound and an anti-CD205 antibody of the invention.

[0238] Various exemplary linkers that may be used with the compositions and methods of the present invention are described in WO 2004 / 010957, U.S. Publication No. 2006 / 0074008, U.S. Publication No. 20050238649, and U.S. Publication No. 2006 / 0024317, each of which is incorporated by reference in its entirety and for all purposes.

[0239] Preferably, the linker is SPDB (N-succinimidyl-3-(2-pyridyldithio)butyrate).

[0240] Pharmaceutical Compositions combination The pharmaceutical combination of the present invention is in the form of a combined preparation for separate or sequential use. Similarly, in the method of the present invention, the components (a) and (b) of the pharmaceutical combination may be administered separately or sequentially.

[0241] The term "pharmaceutical combination" as used in this application refers to a pharmaceutical product that contains at least two active ingredients, either as a single formulation or as separate components.

[0242] The term "combined formulation" as used in this application means a formulation that includes both components a) and b), either as individual components or in a single formulation.

[0243] If the administration is sequential, the delay in administering the second component should be such that the benefit of the effect resulting from using the combination is maximized. Thus, in one embodiment, sequential treatment includes administration of each component of the combination within 84 days. In another embodiment, the period is 77 days. In another embodiment, the period is 70 days. In another embodiment, the period is 63 days. In another embodiment, the period is 56 days. In another embodiment, the period is 49 days. In another embodiment, the period is 42 days. In another embodiment, the period is 35 days. In another embodiment, the period is 28 days. In another embodiment, the period is 24 days. In another embodiment, the period is 21 days. In another embodiment, the period is 18 days. In another embodiment, the period is 15 days. In another embodiment, the period is 13 days. In another embodiment, the period is 11 days. In another embodiment, the period is 9 days or less. In another embodiment, the period is 7 days or less. In another embodiment, the period is 5 days or less. In another embodiment, the period is 3 days or less. In another embodiment, the period is 1 day or less. In a preferred embodiment, the sequential treatment includes administration of each component of the combination within a period of 14-16 days.

[0244] Component (a) is administered first, followed by component (b).

[0245] The ratio of the total amounts of component (a) and component (b) administered in the combined formulation may vary [e.g., to address the needs of a sub-population of patients being treated, or the needs of a single patient (these different needs may be due to the age, sex, weight, etc. of the patient)].

[0246] Components (a) and (b), whether in a single composition or in separate compositions, may be independently formulated with one or more pharma- ceutical acceptable carriers. The pharmaceutical combination of the present invention may also include at least one other anti-tumor agent, or anti-inflammatory or immunosuppressive agent. As used in this application, a "pharma-ceutical acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are physiologically compatible. Preferably, the carrier is suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion). Depending on the route of administration, the active compound, i.e., antibody, immunoconjugate, or bispecific molecule, may be coated with a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.

[0247] Examples of suitable aqueous and non-aqueous carriers that may be used in the combination pharmaceutical of the present invention include, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate). Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

[0248] These combinations or portions thereof may contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the presence of microorganisms can be ensured both by the above sterilization procedures and by the inclusion of various antibacterial and antifungal agents (e.g., paraben, chlorobutanol, phenol sorbic acid, etc.). It may also be desirable to include isotonic agents, such as sugars, sodium chloride, etc., in the composition. Furthermore, the inclusion of agents delaying absorption, such as aluminum monostearate and gelatin, may result in prolonged absorption of the injectable pharmaceutical form.

[0249] Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions, and sterile powders for extemporaneous preparation of sterile injectable solutions or dispersions.The use of such media and agents for pharmaceutically active substances is known in the art.The use of any conventional media or agent in the pharmaceutical composition of the present invention is contemplated except insofar as it is incompatible with said active compound.Auxiliary active compounds can also be incorporated into said composition.

[0250] Sterile injectable solution may be prepared by incorporating the required amount of active compound into a suitable solvent with one or a combination of the above-listed ingredients, and then optionally sterilizing by microfiltration.Generally, dispersion is prepared by incorporating active compound into a sterile medium that contains a basic dispersion medium and other necessary ingredients from the above-listed ones.In the case of sterile powder for preparing sterile injectable solution, the preferred method of preparation is vacuum drying and lyophilization (freeze drying), which produces a powder of active ingredient + any additional desired ingredients from the solution that has been previously sterilized and filtered.

[0251] The amount of active ingredient that can be combined with carrier material to produce a single dosage form varies depending on the subject to be treated and the specific mode of administration.The amount of active ingredient that can be combined with carrier material to produce a single dosage form is generally the amount of said composition that produces a therapeutic effect.Generally, this amount will be in the range of about 0.01% to about 99% active ingredient, preferably about 0.1% to about 70%, most preferably about 1% to about 30% active ingredient, of 100%, combined with pharmaceutically acceptable carrier.

[0252] The dosage regimen is adjusted to provide the optimum desired response (e.g., therapeutic response). For example, it may be administered in a single bolus, or several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. For ease of administration and uniformity of dosage, it is particularly useful to formulate the parent composition into dosage units. Dosage unit form, as used in this application, refers to physically discrete units suitable as unitary dosages for the subjects to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specific details of the dosage unit forms of the present invention are determined by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such active compounds to treat sensitivities in individuals.

[0253] For administration of anti-CD205-DM4 ADCs, the dosage ranges from about 0.8 to 10 mg / kg of host body weight, e.g., 1.0 mg / kg to 8.0 mg / kg, 1.2 mg / kg to 7.5 mg / kg, 1.4 mg / kg to 7.0 mg / kg, 1.6 to 6.0 mg / kg, 1.6 to 5 mg / kg, 2.0 to 4 mg / kg, 2.5 to 3.6 mg / kg. For example, the dosage can be 0.8 mg / kg, 1.0 mg / kg, 1.2 mg / kg, 1.4 mg / kg, 1.6 mg / kg body weight, 2.0 mg / kg body weight, 2.5 mg / kg body weight, 3.5 mg / kg body weight, 4 mg / kg body weight, or 5 mg / kg body weight. Exemplary treatment regimens entail administration once per week, once per two weeks, once per three weeks, once per four weeks, once per month, once per six weeks, once per three months, or once per three to six months.

[0254] Preferred dosing regimens of anti-CD205-DM4 ADC for use in the methods of the invention include 2.0 mg / kg body weight, 2.5 mg / kg body weight, 3.0 mg / kg body weight or 3.5 mg / kg body weight by intravenous administration, with the antibody drug conjugate being administered using one of the following dosing schedules: (i) six doses every three weeks; (ii) every three weeks; (iii) one dose of 2.5 mg / kg body weight followed by 2 mg / kg body weight every three weeks.

[0255] Further preferred dosing regimens for anti-CD205 antibody drug conjugates for use in the methods of the invention include 0.8 mg / kg body weight, 1.0 mg / kg body weight, 1.2 mg / kg body weight or 1.4 mg / kg body weight by intravenous administration, wherein the antibody drug conjugate is administered using one of the following dosing schedules: (i) once per week; (ii) once per week for four doses; (iii) once per week for three doses; (iv) three times per week, once every three weeks.

[0256] For administration of PD1 antibodies, dosages are from 200 mg to 480 mg, e.g., 200 mg, 240 mg, 400 mg, or 480 mg. Exemplary treatment regimens entail administration once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, or once every 6 weeks.

[0257] For administration of PD-L1 antibodies, the dosage is from 800 mg to 1500 mg, for example, 800 mg, 1200 mg or 1500 mg. Exemplary treatment regimens entail administration once every two weeks, once every three weeks, or once every four weeks.

[0258] In some methods, two or more monoclonal antibodies with different binding specificities are administered simultaneously, in which case the dosage of each antibody administered falls within the ranges indicated.

[0259] The actual dosage level of the active ingredient in the pharmaceutical combination of the present invention can be varied for a specific patient, composition, and mode of administration to obtain an amount of the active ingredient effective to achieve the desired therapeutic response without causing toxicity to the patient. The selected dosage level depends on various pharmacokinetic factors, such as, for example, the activity of the specific composition of the present invention used, or its ester, salt, or amide, the route of administration, the timing of administration, the excretion rate of the specific compound used, the duration of treatment, other drugs, compounds and / or substances used in combination with the specific composition used, the age, sex, weight, symptoms, general health and previous medical history of the patient being treated, and factors well known in the medical art.

[0260] A "therapeutically effective dosage" of an anti-CD205 antibody or combination of the invention preferably results in a decrease in the severity of disease symptoms, an increase in the frequency and duration of disease symptom-free periods, or prevention of functional impairment or disability due to disease affliction. For example, for treating CD205 or PD1 / PD-L1 mediated tumors, a "therapeutically effective dosage" preferably inhibits cell proliferation or tumor growth by at least about 20%, at least about 30%, more preferably at least about 40%, at least about 50%, even more preferably at least about 60%, at least about 70%, even more preferably at least about 80%, or at least about 90%, compared to untreated subjects. The ability of a compound to inhibit tumor growth may be assessed in an animal model system predictive of efficacy in human tumors. Alternatively, this property of a composition may be assessed by testing the ability of the compound to inhibit cell proliferation, and such inhibition may be measured in vitro by assays known to those of skill in the art. A therapeutically effective amount of a therapeutic compound may reduce tumor size or otherwise alleviate symptoms in a subject. One of ordinary skill in the art would be able to determine such an amount based on factors such as the size of the subject, the severity of the subject's symptoms, and the particular composition or route of administration selected.

[0261] The pharmaceutical combination of the present invention may be administered via one or more routes of administration using one or more of a variety of methods known in the art. Components (a) and (b) may be administered by the same route or by different routes. As will be appreciated by those skilled in the art, the route and / or mode of administration will vary depending on the desired results. Preferred routes of administration of the antibody of the present invention include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes, for example, by injection or by infusion. The phrase "parenteral administration" as used in this application means a mode of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion.

[0262] Alternatively, the antibody may be administered by a non-oral route, such as a topical, epidermal or mucosal route of administration, such as intranasal, oral, vaginal, rectal, sublingual or topical.

[0263] The active compound may be prepared with a carrier that will protect the compound against rapid release, such as a controlled release formulation, such as implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers may be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for preparing such formulations are patented or generally known to those skilled in the art [see, for example, Sustained and Controlled Release Drug Delivery Systems (1978) JR Robinson, ed., Marcel Dekker, Inc., NY].

[0264] Therapeutic compositions may be administered using medical devices known in the art. For example, in preferred embodiments, the antibody or antibodies may be administered using a needleless hypodermic injection device, such as those disclosed in U.S. Patent Nos. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556. Examples of well-known implants and modules useful in the present invention include: U.S. Patent No. 4,487,603, which discloses an implantable microinfusion pump for dispensing drugs at a controlled rate; U.S. Patent No. 4,486,194, which discloses a therapeutic device for administering medication through the skin; U.S. Patent No. 4,447,233, which discloses a drug infusion pump for delivering drugs at a precise infusion rate; U.S. Patent No. 4,447,224, which discloses a variable flow implantable infusion device for continuously delivering drugs; U.S. Patent No. 4,439,196, which discloses an osmotic drug delivery system having a multi-chamber compartment; and U.S. Patent No. 4,475,196, which discloses an osmotic drug delivery system. These patents are incorporated by reference into this application. Many other such implants, delivery systems, and modules are known to those of skill in the art.

[0265] In certain embodiments, the anti-CD205 and / or anti-PD1 / PD-L1 antibodies may be formulated to ensure proper distribution in vivo. For example, the blood-brain barrier (BBB) ​​excludes many highly hydrophilic compounds. To ensure that therapeutic compounds cross the BBB (if desired), they may be formulated, for example, in liposomes. For methods of making liposomes, see, for example, U.S. Patent Nos. 4,522,811; 5,374,548; and 5,399,331. The liposomes may contain one or more moieties that are selectively transported to specific cells or organs, thus enhancing targeted drug delivery [see, for example, VV Ranade (1989) J. Clin. Pharmacol. 29:685]. Exemplary targeting moieties include folate or biotin (see, e.g., U.S. Pat. No. 5,416,016); mannosides [Umezawa et al. (1988) Biochem. Biophys. Res. Commun. 153:1038]; antibodies [PG Bloeman et al. (1995) FEBS Lett. 357:140; M. Owais et al. (1995) Antimicrob. Agents Chemother. 39:180]; surfactant protein A receptor [Briscoe et al. (1995) Am. J. Physiol. 1233:134]:p120 [Schreier et al. (1994) J. Biol. Chem. 269:9090]; K. Keinanen; ML Laukkanen (1994) FEBS Lett. 346:123; JJ Killion; see also IJ Fidler (1994) Immunomethods 4:273.

[0266] Uses and Methods As used in this application, the term "subject" is intended to include humans and non-human animals. Non-human animals include all vertebrates, e.g., mammals and non-mammals, e.g., non-human primates, sheep, dogs, cats, cows, horses, chickens, amphibians, and reptiles. Preferred subjects include human patients with disorders mediated by CD205 activity and / or PD1 / PD-L1 activity. Suitable routes for administering the antibody compositions (e.g., monoclonal antibodies, and immunoconjugates) in vivo and in vitro are well known in the art and may be selected by the skilled artisan. For example, the antibody compositions may be administered by injection (e.g., intravenously or subcutaneously). Appropriate dosages for the molecules used depend on the age and weight of the subject, as well as the concentration and / or formulation of the antibody composition.

[0267] As mentioned above, the anti-CD205 and / or anti-PD1 / PD-L1 antibodies may be co-administered with one or more other therapeutic agents, such as cytotoxic, radiotoxic or immunosuppressive agents. The antibodies may be linked to the agents (as an immunoconjugate) or may be administered separately from the agents. In the latter case (administration separately), the antibodies may be administered before, after or simultaneously with the agents, or may be co-administered with other known therapies, such as anti-cancer therapies, such as radiation. Such therapeutic agents include, inter alia, anti-neoplastic agents. Other agents suitable for co-administration with the antibodies of the invention include other agents used to treat cancer (e.g. gastric, endometrial, colorectal, prostate, breast, ovarian or lung cancer). Co-administration of the anti-CD205 antibodies or antigen-binding fragments thereof of the invention together with chemotherapeutic agents provides two anti-cancer agents that act through different mechanisms to produce a cytotoxic effect on human tumor cells. Such co-administration can overcome problems due to the development of resistance to the drugs or changes in the antigenicity of tumor cells, which renders them unresponsive to the antibodies.

[0268] The pharmaceutical combination of the invention may also be administered together with serum and / or complement. These compositions may be advantageous when the complement is located in close proximity to the antibody. Alternatively, the antibody and the complement or serum may be administered separately.

[0269] Also within the scope of the invention is a kit comprising components (a) and (b) together with instructions for use. The kit may further contain one or more additional reagents, such as an immunosuppressant, cytotoxic agent, or radiotoxic agent, or one or more additional antibodies, such as an antibody that binds to an epitope in the CD205 antigen that is distinct from the first antibody and has complementary activity.

[0270] Thus, a patient treated with a pharmaceutical combination of the invention may additionally be administered (before, simultaneously with, or after administration of an antibody disclosed in the present application) another therapeutic agent (e.g., a cytotoxic agent or a radiotoxic agent) that enhances or potentiates the therapeutic effect of the antibody.

[0271] All references cited herein, including, but not limited to, all articles, publications, patents, patent applications, presentations, texts, reports, manuscripts, brochures, books, internet postings, journal articles, periodicals, product fact sheets, etc., are hereby incorporated by reference in their entirety. The discussion of references in this application is intended merely to summarize the assertions made by their authors and is not an admission that any reference is prior art, and applicants reserve the right to challenge the accuracy and pertinence of the cited references.

[0272] Although the foregoing invention has been described in some detail by way of illustration and example for clarity of understanding, it will be readily apparent to those skilled in the art in light of the teachings of this invention that certain changes and modifications can be made without departing from the spirit or scope of the appended claims.

[0273] This invention is further illustrated by the following examples which should not be construed as further limiting. EXAMPLES

[0274] Working Example Example 1: Generation of human monoclonal antibodies against the CD205 antigen Mice (xenogeneic mouse IgG1) were immunized with CHO cells transfected with full length CD205 according to standard methods.

[0275] The specificity of the antibodies raised against CD205 was tested by flow cytometry on CD205 transfected HEK293 cells and subsequently on CD205-expressing HT29 cells. To test the ability of the antibodies to bind to cell surface CD205 protein, the antibodies were incubated with CD205-expressing cells. Cells were washed with FACS buffer (DPBS, 2% FBS), centrifuged, and resuspended in 100 μl of diluted primary CD205 antibody (diluted in FACS buffer). The antibody-cell line complex was incubated on ice for 60 minutes and then washed twice with FACS buffer as above. The cell-antibody pellet was resuspended in 100 μl of diluted secondary antibody (diluted in FACS buffer) and incubated on ice for 60 minutes. The pellet was washed as before and resuspended in 200 μl of FACS buffer. The samples were loaded onto a BD FACScanto II flow cytometer and the data were analyzed using BD FACSdiva software (results not shown).

[0276] Example 2: Structural characterization of monoclonal antibodies against CD205 The cDNA sequences encoding the heavy and light chain variable regions of the CD205_A1 monoclonal antibody were obtained using standard PCR techniques and sequenced using standard DNA sequencing techniques.

[0277] The antibody sequence may be subjected to mutagenesis at one or more residues back to germline residues.

[0278] The nucleotide and amino acid sequences of the heavy chain variable region of CD205_A1 are shown in SEQ ID NOs: 3 and 1, respectively.

[0279] The nucleotide and amino acid sequences of the light chain variable region of CD205_A1 are shown in SEQ ID NOs: 4 and 2, respectively.

[0280] Further analysis of the CD205_A1 VH sequence using the Kabat system for CDR region determination delineates the heavy chain CDR1, CDR2 and CDR3 regions, as shown as SEQ ID NOs: 5, 6 and 7, respectively. The CD205_A1 CDR1, CDR2 and CDR3 VH sequences are shown in FIG.

[0281] Further analysis of the CD205_A1 VK sequence using the Kabat system for CDR region determination delineates the light chain CDR1, CDR2 and CDR3 regions, as shown as SEQ ID NOs: 8, 9 and 10, respectively. The CD205_A1 CDR1, CDR2 and CDR3 VK sequences are shown in FIG.

[0282] Example 3: Efficacy of various DM4-conjugated anti-LY75 monoclonal antibodies in Raji and THP1 cells THP-1 and Raji cells were prepared at a seeding density of 3,000 cells / well (1.5×10 5 cells / mL) and added to the assay plates (20 μL / well).

[0283] THP-1 cells were prepared in RPMI GLUTAMAX Growth (2ME) and Raji cells were prepared in RPMI 1640 ATCC Growth AB-Free (10%).

[0284] Each conjugated antibody was prepared in triplicate at a starting concentration that was twice the final concentration and diluted to the final concentration in RPMI 1640 ATCC Growth AB-Free (10%). Antibodies were transferred to the required assay plates and incubated for 96 hours.

[0285] Following assay incubation, Cell-Titer Glo was added to each plate and read using a plate reader set for luminescence with 0.2 second integration.

[0286] The raw data was converted to % specific killing (data not shown) using the negative control (target cells only) and EC50 was calculated. The EC50 of the antibodies against the two cell lines is shown in Table 3. As can be seen from Table 3, the antibody conjugate CD205_A1 showed a lower EC50 than the other two antibodies tested. However, all three conjugates showed cytotoxicity against both Raji and THP1 cells. [Table 3]

[0287] Example 4: Toxicity of DM1-conjugated and DM4-conjugated anti-CD205 monoclonal antibodies in cynomolgus monkeys Six male monkeys were assigned to this study, with 2 monkeys per group. Either vehicle (PBS), CD205_DM4 (cleavable) or CD205_DM1 (non-cleavable) was administered twice (days 1 and 29) by 15-minute intravenous infusion at 0 mg / kg / dose (PBS, vehicle), 5 mg / kg / dose (CD205_DM4, cleavable) or 10 mg / kg / dose (CD205_DM1, non-cleavable). Blood samples were taken for toxicokinetic evaluation before administration began (day 1) and 1, 2, 3, 7, 14, 21 and 28 days after each administration (day 28 after the first administration was also the pre-dose time point for the second administration). Blood samples were taken for clinical pathology analysis before administration began (day 1) and 1, 3, 7, 14, 21 and 28 days after each administration (day 28 after the first administration was also the pre-dose time point for the second administration). All study animals were euthanized and necropsied after the final blood draw on day 57. Separated plasma from each blood draw was isolated, frozen, and shipped to Oxford BioTherapeutics, Inc. and analyzed for ADC concentrations by ELISA.

[0288] Treatment-related clinical pathology findings included mild regenerative anemia and a transient decrease in the blood white blood cell profile, especially the neutrophil count. Anemia was observed in both animals treated with CD205_DM4 at 5 mg / kg and in one of two animals treated with CD205_DM1 at 10 mg / kg. Severe neutropenia was observed in all animals, with counts reaching nadirs one week after treatment and rapidly recovering; absolute neutrophil count nadirs were lower in animals treated with CD205_DM4. No test article-related effects on APTT and PT coagulation parameters were observed. Serum chemistry changes included transient increases in AST, CK, LDH (in one of two animals in each treatment group) and globulins after treatment with CD205_DM4 at 5 mg / kg and CD205_DM1 at 10 mg / kg. Furthermore, a transient increase in the liver-specific enzyme ALT was observed only in CD205_DM4-treated animals. The short-term and / or magnitude of increases in serum chemistry parameters suggests that they are not adverse. There were no test substance-related urinary test findings. Necropsy examination after a 4-week recovery period revealed no treatment-related gross pathology findings or absolute and relative organ weight changes. Only histopathological findings in the thyroid (changes in colloid morphology of follicles) and kidney (dilated tubules in the outer cortex) were assessed as minimal severity; not associated with changes in other study parameters; and of non-adverse and small toxicological significance. Conclusion: Repeated treatment at two doses, 5 mg / kg CD205_DM4 or 10 mg / kg CD205_DM1, was well tolerated in cynomolgus monkeys. All treatment-related toxicological findings were reversible after a 4-week recovery period.

[0289] Example 5: Immunohistochemistry protocol for CD205. CD205 target expression levels are assessed in formalin-fixed paraffin-embedded (FFPE) human tumors using immunohistochemistry (IHC) staining assays. FFPE tissues were sectioned at 4-6 microns thickness on a rotary microtome and mounted onto positively charged glass slides. The mounted sections were air-dried on the slides overnight at room temperature or for 30 min at 37°C, followed by baking at 60°C for 30 min. The slides were deparaffinized in three changes of xylene for 5 min each and rehydrated through graded ethanol (first three changes of 100% ethanol, followed by one change of 95% ethanol and one change of 80% ethanol) and two rinses in deionized water, for 3 min each. After the deparaffinization and rehydration process, the slides underwent heat-induced epitope retrieval (HIER) in Diva Decloaker solution (DDV2004) in a Biocare Decloaker NxGen pressure cooker. The slides were exposed to a temperature of 110°C for 15 minutes and cooled in the unit for another 10 minutes before removal. After removal from the pressure cooker, the slides were equilibrated to room temperature by slowly replacing the hot Diva retrieval solution with deionized or distilled water. The slides were rinsed with Tris-Buffered Saline (TBS) (TWB945) and loaded into the staining rack of an intelliPATH automated stainer (IPS0001US). The slides were incubated in 300ul of Peroxidazed 1 (PX968) for 5 minutes to block endogenous peroxidase. The Peroxidazed 1 was then removed and the slides were incubated in 300 ul of Background Punisher (IP974G20) for 10 minutes to block non-specific protein-protein interactions. The slides were then washed with TBS and primary antibodies were applied.The primary antibody was a mouse monoclonal antibody against CD205 supplied by Leica Biosystems (cat. no. NCL-L-CD205) and was used diluted 1:80 (0.5 ug / mL) in Da Vinci Green Diluent (PD900). 300ul of diluted primary antibody was applied to the slides and incubated at room temperature for 30 minutes. After incubation with the primary antibody, the slides were washed in TBS and 300ul of secondary detection antibody polymer MACH 2 mouse HRP (MHRP520) was applied and incubated at room temperature for 30 minutes. The slides were washed in TBS and developed in 300ul of intelliPATH FLX DAB chromogen for 5 minutes. After the chromogen was developed, the slides were washed in deionized or distilled water, lightly counterstained with hematoxylin for 20 seconds, and rinsed again with deionized water. The stained slides were then dehydrated through graded histological grade ethanol changes (3-5 min each): 70%, 90%, 95%, three times 100%, and three changes of xylene, and then mounted in Permount.

[0290] Staining was scored on a scale of 0 (negative) to 3+ (highly positive), with 1+ being low positivity and 2+ being moderately positive. Pathology graders assessed and recorded the percentage of tumor cells showing membranous staining at each intensity level (e.g., 0=5%, 1+=50%, 2+=35%, 3+=10%).

[0291] Patients exhibiting greater than 50% CD205 tumor expression that was at least 2+ were selected as suitable for treatment with CD205-DM4 ADC. For the avoidance of doubt, the antibody portion of CD205-DM4-ADC includes the antibody CD205_A1.

[0292] Example 6: Effect of anti-CD205 DM4 ADC on T-cell populations in the blood of gastric cancer patients Patients with metastatic gastric cancer were administered CD205-DM4 ADC at a dosage of 2.5 mg / kg (day 0). Blood was collected from gastric cancer patients 1, 8, 15, and 21 days after treatment.

[0293] method All steps were performed at room temperature. 100 μl of patient blood was dispensed into each microcentrifuge tube and antibodies were added at the appropriate concentrations (see table). The blood samples were stained for 20 min at room temperature and 1 ml of 1× RBC lysis buffer was added. The cells were incubated for an additional 15 min and centrifuged at 300 g for 5 min. The buffer was removed and the pellet was washed with 1 ml of FACS staining buffer (2% FCS + PBS + 0.05% sodium azide).

[0294] The pellet was resuspended in 500-700 μl of FACS buffer and the samples were analyzed by FACS analysis. [Table 4]

[0295] FACS gating strategy Lymphocytes were first isolated from blood using CD45-PE antibody. The T-cells were then separated using CD3-PerCp-Cy5.5 antibody. Separate populations of CD4+ and CD8+ cells were separated using CD4-PECY7 and CD8-FITC, respectively. CD4+ and CD8+ cells were then screened for CD205 and PD1 expression using CD205-Alexa Fluor 647 and PD1-BV421.

[0296] result In FIG. 3, the left panel shows a 3-fold increase in the number of CD8+ T-cells present in the patient's blood between days 8 and 21 of a 21-day time series after administration of the CD205-DM4 ADC drug. The right panel shows a 3.4-fold increase in the number of CD4+ T-cells present in the patient's blood between days 8 and 21 of a 21-day time series after administration of the CD205-DM4 ADC drug. As can be seen, the numbers of CD8+ and CD4+ T-cells remain relatively constant until day 15. Thereafter, T-cell levels rapidly increase approximately 3-fold between days 15 and 21.

[0297] FIG. 4 shows in the left panel that the proportion of CD4+ and CD8+ T-cells relative to the total T-cell population remained relatively constant over time.

[0298] The right panel shows the percentage of CD4+ and CD8+ T cells that were also PD1+. As can be seen for both CD4+ and CD8+, the percentage of PD1 positive T-cells rose rapidly from day 8 onwards, peaking at day 15.

[0299] Figure 5 shows in the left panel the evolution over time of the number of CD8+ T-cells present in the patient's blood that are also PD1+. The right panel shows the evolution over time of the number of CD4+ T-cells present in the patient's blood that are also PD1+. As can be seen, the number of CD8+ PD1+ T-cells initially decreases slightly, but then increases approximately four-fold from day 8 to day 21. A similar pattern is seen for CD4+ PD1+ T-cells.

[0300] In contrast, Figures 6 and 7 show that the populations of CD8+CD205+ and CD4+205+ immune cells were dramatically reduced to very low levels by day 8 and did not recover even at day 21.

[0301] It has been previously reported that CD8+CD205+ immune cells can induce Foxp3+ regulatory T cells, which are known to mediate immunological self-tolerance and suppress immune responses (Yamazaki, S; et al, J. Immunol., 181(10), 6923,

[2008] ).

[0302] conclusion The increase in the number of T-cells one week after CD205-DM4 ADC-induced depletion of CD4+ CD205+ and CD8+ CD205+ immunoregulatory cells supports the use of CD205-DM4 ADC as a therapeutic modality to reactivate the suppressed immune system of patients to induce an immune response against tumors. Furthermore, the increase in the number of PD1+ T-cells after treatment with CD205-DM4 ADC supports the prevention of the CD205-DM4 ADC-induced immune response from being subsequently blocked by tumors by the use of the immune checkpoint inhibitor PD1 / PD-L1.

[0303] Example 7: Effect of anti-CD205 DM4 ADC on dendritic cell populations in the blood of gastric cancer patients method All steps were performed at room temperature. 100 μl of patient blood was dispensed into each microcentrifuge tube and the appropriate antibody was added (see table). The blood samples were stained for 20 min at room temperature and 1 ml of 1× RBC lysis buffer was added. The cells were incubated for an additional 15 min and centrifuged at 300 g for 5 min. The buffer was removed and the pellet was washed with 1 ml of FACS staining buffer (2% FCS + PBS + 0.05% sodium azide).

[0304] The pellet was resuspended in 500-700 μl of FACS buffer and the samples were analyzed by FACS analysis. [Table 5]

[0305] FACS gating strategy Dendritic cells were first isolated from blood using HLA-DR FITC and Lineage-BV510 antibodies. The dendritic cells were then separated into pDCs and mDCs using CD11c (mDC) and CD123 (pDC) antibodies. Separate populations of mDCs and pDCs were then screened for CD205 and PD-L1 expression using CD205-Alexa Fluor 647 and PD-L1-PE.

[0306] result In Figure 8, the top left panel shows that the total number of mDCs in peripheral blood increased 4.5-fold over the 21-day timeline following administration of the drug. The bottom left panel shows that after the initial drop, the total number of peripheral pDCs doubled over the 21-day timeline following administration of the drug. The right panel shows a similar pattern for CD205+ mDCs and pDCs, with a sharp rise between days 8 and 21 following the initial drop in CD205.

[0307] Example 8: Clinical response of gastric cancer patients to treatment with 2.0-2.5 mg / kg CD205-DM4 ADC A patient with chemotherapy-refractory advanced gastric cancer whose tumor was MSI stable, PD-L1 negative, and had previously received two lines of chemotherapy (1st line docetaxel / cisplatin / 5FU; 2nd line ramucirumab / paclitaxel) and progressed, and had lymph node metastasis and malignant ascites, was screened by IHC for CD205 tumor expression. IHC showed that the primary tumor showed 60% 2+ CD205 expression, which met the criteria for treatment (data not shown). The patient was treated with CD205-DM4 ADC administered at 2.5 mg / kg in a 21-day cycle. After the first cycle, the dose was reduced to 2.0 mg / kg. After three cycles of treatment, the patient was evaluated. It was shown that the primary gastric tumor had shrunk by about 40%, and the lymph node metastasis had disappeared, as had the ascites (see Table 5). Two further cycles of CD205-DM4 ADC were administered, followed by one cycle of pembrolizumab (200 mg) (approximately 4 weeks after the last cycle of CD205-DM4 ADC). After treatment with pembrolizumab, clinical examination revealed a complete response to the primary gastric tumor. [Table 6]

[0308] Example 9: Patient Blood Sample Analysis A blood sample taken from a gastric cancer patient (Patient 1) on day 1 of cycle 1 was analyzed for CD205+ expression. In this patient, both CD4+ and CD8+ T-cells were found to express CD205 at high levels (see Figure 6).

[0309] Furthermore, an esophageal cancer patient (patient 2) who received CD205 DM4 ADC and developed stable disease (data not shown) also demonstrated high levels of CD205 expression on both CD4+ and CD8+ T-cells isolated from a blood sample taken on day 1 of cycle 1 of treatment.

[0310] Patients 3-5 showed low levels of expression of CD205 on CD4+ and CD8+ T-cells and did not respond in the same way as patients 1 and 2.

[0311] Furthermore, an endometrial cancer patient (patient 6) who demonstrated a complete response after two cycles of treatment with the CD205 DM4 ADC and one cycle of treatment with pembrolizumab was shown to have high levels of CD205 expression on both CD4+ and CD8+ T-cells. [Table 7]

[0312] Given the association between high levels of CD205+ T-cells in the blood of cancer patients and the anti-tumor efficacy of treatment with CD205-DM4 ADC, this measurement can be used to select patients suitable for treatment with said therapy.

[0313] Example 10: Clinical response of endometrial cancer patients to treatment with 3.0 mg / kg CD205-DM4 ADC A patient with advanced endometrial cancer (Patient 6 above) with stable MSI tumors, low PD-L1 expression (TPS 10%; not eligible for CPI treatment), and who had previously received two lines of chemotherapy (1st line carboplatin / taxol / herceptin; 2nd line letrozole / everolimus) and had progressed lung and liver metastases was screened by IHC for CD205 tumor expression. IHC showed that the primary tumor showed 100% 3+ CD205 expression, which met the criteria for treatment (data not shown). The patient was treated with CD205-DM4 ADC administered at 3 mg / kg in 21-day cycles. After two cycles of treatment, the patient was administered one cycle of pembrolizumab (200 mg) (~3 weeks after the last cycle of CD205-DM4 ADC). Following treatment with pembrolizumab, the patient experienced a complete response to the primary endometrial tumor, as well as liver and lung metastases.

[0314]

Table 8-1

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Claims

1. Combination medications including: iii) an anti-CD205 antibody, or an antigen-binding portion thereof, wherein the antibody comprises: A heavy chain variable region comprising: iii) a first vhCDR comprising SEQ ID NO: 5; ii) a second vhCDR comprising SEQ ID NO:6; iii) a third vhCDR comprising SEQ ID NO: 7; and A light chain variable region comprising: iii) a first vlCDR comprising SEQ ID NO: 8; ii) a second vlCDR comprising SEQ ID NO:9; iii) a third vlCDR comprising SEQ ID NO: 10; and b) Checkpoint modulators.

2. The pharmaceutical combination according to claim 1, wherein said pharmaceutical combination is in the form of a combined preparation for simultaneous, separate or sequential use, preferably sequential use.

3. The pharmaceutical combination according to claim 1 or claim 2, wherein the checkpoint modulator is a PD1 / PD-L1 inhibitor, preferably, the PD1 / PD-L1 inhibitor is an antibody.

4. 4. The pharmaceutical combination of claim 3, wherein the PD1 / PD-L1 inhibitor is selected from the group consisting of nivolumab (MDX-1 106, Opdivo; Bristol-Myers Squibb), pembrolizumab (MK-3475, Keytruda, lambrolizumab, BMS-936558; Merck), dostallimab (TSR-042, Tesaro, Inc.), cemiplimab (REGN-2810, Libtayo; Regeneron), EH12.2H7 (BioLegend, Cat. No. 329902), Balstilimab (Agenus Inc.), avelumab (Bavencio; EMD Serono, Pfizer), durvalumab (Imfinzi, AstraZeneca), BMS-936559, atezolizumab (Tecentriq, Genentech), or their equivalents.

5. The pharmaceutical combination according to claim 1 or 2, wherein the anti-CD205 antibody or antigen-binding portion thereof comprises a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 1, and a light chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO:

2.

6. 3. The pharmaceutical combination according to claim 1 or 2, wherein the antibody that binds to CD205 comprises: (i) a heavy chain having at least 80%, 85%, 90%, 95%, 99% or 100% amino acid sequence identity to SEQ ID NO:100; and (ii) a light chain having at least 80%, 85%, 90%, 95%, 99% or 100% amino acid sequence identity to SEQ ID NO:

101.

7. The pharmaceutical combination according to claim 1 or 2, wherein the antibody or antigen-binding portion thereof further comprises a covalently linked moiety.

8. The pharmaceutical combination according to claim 7, wherein the moiety is a drug.

9. 9. The pharmaceutical combination of claim 8, wherein the drug is selected from the group consisting of maytansinoids, dolastatins, hemiasterlins, auristatins, trichothecenes, calicheamicins, duocarmycins, bacterial immunotoxins, pyranoindoidinoquinolines, camptothecins, anthracyclines, antheamicins, thienoindoles, amatoxins, CC1065 or taxol, and derivatives thereof.

10. 10. The pharmaceutical combination according to claim 9, wherein the drug is a maytansinoid selected from the group consisting of DM4 and DM1, preferably DM4.

11. 3. The pharmaceutical combination according to claim 1 or 2, wherein the pharmaceutical combination comprises at least one pharma- ceutically acceptable diluent, excipient, or carrier.

12. A pharmaceutical combination for use in treating or preventing cancer, wherein the pharmaceutical combination comprises: an antibody or antigen-binding portion thereof that regulates a population of CD205+ immunoregulatory cells; and a composition comprising a checkpoint modulator.

13. The pharmaceutical combination for use according to claim 12, wherein the checkpoint modulator is a PD1 or PD-L1 inhibitor, preferably PD1.

14. A pharmaceutical combination for use in enhancing the effect of an inhibitor of PD-1 / PD-L1 interaction in a patient, wherein the pharmaceutical combination comprises: an antibody, or antigen-binding portion thereof, that modulates a population of CD205+ immunoregulatory cells; and a composition comprising an inhibitor of PD1 / PD-L1 interaction.

15. A pharmaceutical combination for use according to any one of claims 12 to 14, wherein the pharmaceutical combination is in the form of a combined preparation for simultaneous, separate or sequential use, preferably sequential use.

16. 15. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the patient is refractory to at least one chemotherapy or the patient's cancer is progressing during at least one chemotherapy.

17. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the patient is refractory to checkpoint modulator therapy.

18. 18. The pharmaceutical combination for use according to claim 17, wherein the checkpoint modulator therapy is a PD1 inhibitor therapy.

19. The combination drug for use according to any one of claims 12 to 14, wherein the cancer is PDL1 negative or low.

20. 15. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the cancer is MSI stable.

21. A pharmaceutical combination for use according to any one of claims 12 to 14, wherein the antibody or antigen-binding portion thereof binds to CD205.

22. 15. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the antibody or antigen-binding portion thereof that binds to CD205 comprises: A heavy chain variable region comprising: i) a first vhCDR comprising SEQ ID NO:5; ii) a second vhCDR comprising SEQ ID NO:6; and iii) a third vhCDR comprising SEQ ID NO: 7; and A light chain variable region comprising: i) a first vlCDR comprising SEQ ID NO: 8; ii) a second vlCDR comprising SEQ ID NO: 9; and iii) a third vlCDR comprising SEQ ID NO: 10 wherein, optionally, any one or more of the above SEQ ID NOs independently comprise one or two amino acid substitutions, preferably conservative substitutions.

23. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the antibody or antigen-binding portion thereof comprises a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO: 1, and a light chain variable region having at least 80%, 85%, 90%, 95% or 99% amino acid sequence identity to SEQ ID NO:

2.

24. 15. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the antibody binding to CD205 comprises: (i) a heavy chain having at least 80%, 85%, 90%, 95%, 99% or 100% amino acid sequence identity to SEQ ID NO:100; and (ii) a light chain having at least 80%, 85%, 90%, 95%, 99% or 100% amino acid sequence identity to SEQ ID NO:

101.

25. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the antibody or antigen-binding portion thereof further comprises a covalently linked moiety.

26. 26. The pharmaceutical combination for use according to claim 25, wherein the moiety is a drug.

27. 27. The pharmaceutical combination for use according to claim 26, wherein the drug is selected from the group consisting of maytansinoids, dolastatins, hemiasterlins, auristatins, trichothecenes, calicheamicins, duocarmycins, bacterial immunotoxins, pyranoindoidinoquinolines, camptothecins, anthracyclines, antheamicins, thienoindoles, amatoxins, CC1065 or taxol, and derivatives thereof.

28. 28. The pharmaceutical combination for use according to claim 27, wherein the drug is a maytansinoid selected from the group consisting of DM4 and DM1, preferably DM4.

29. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein said checkpoint modulator is an antibody.

30. 30. The pharmaceutical combination for use according to claim 29, wherein the antibody is an anti-PD1 or PD-L1 antibody.

31. 31. The pharmaceutical combination for use according to claim 30, wherein the anti-PD-1 antibody is nivolumab (MDX-1 106, Opdivo; Bristol-Myers Squibb), pembrolizumab (MK-3475, Keytruda, lambrolizumab, BMS-936558; Merck), dostallimab (TSR-042, Tesaro, Inc.), cemiplimab (REGN2810, Regeneron Pharmaceuticals), EH12.2H7 (BioLegend, Cat. No. 329902), Balstilimab (Agenus Inc.).

32. 31. The pharmaceutical combination for use according to claim 30, wherein the anti-PD-L1 antibody is avelumab (Bavencio; EMD Serono, Pfizer), durvalumab (Imfinzi, AstraZeneca), BMS-936559, atezolizumab (Tecentriq, Genentech).

33. 31. A pharmaceutical combination for use according to claim 30, comprising: administering the antibody, or antigen-binding portion thereof, once per week or once per three weeks; and The anti-PD1 or PD-L1 antibody is administered once every three weeks.

34. An antibody or an antigen-binding portion thereof, or a combination pharmaceutical for the use according to claims 12 to 14, wherein the cancer is a CD205-positive cancer.

35. 15. The pharmaceutical combination for use according to any one of claims 12 to 14, wherein the cancer is selected from the group consisting of gastric cancer, endometrial cancer, esophageal cancer, lung cancer, ovarian cancer, gastroesophageal junction cancer, pancreatic cancer, breast cancer, colorectal cancer, skin cancer, thyroid cancer, kidney cancer, and the like. cancer), liver cancer, head and neck cancer, bladder cancer, leukemia, preferably acute myeloid leukemia or chronic lymphocytic leukemia, myeloma, preferably multiple myeloma and lymphoma, preferably diffuse large B-cell lymphoma (DLBCL), B-cell lymphoma, follicular lymphoma, mantle cell lymphoma, lymphoma of mucosa-associated lymphoid tissue (MALT), T-cell / histiocyte-rich B-cell lymphoma, Burkitt's lymphoma, lymphoplasmacytic lymphoma, small lymphocytic lymphoma, marginal zone lymphoma, T-cell lymphoma, peripheral T-cell lymphoma, anaplastic large cell lymphoma and angioimmunoblastic T-cell lymphoma.

36. 36. The antibody or antigen-binding portion thereof, or the combination pharmaceutical for use according to claim 35, wherein the cancer is selected from the group comprising gastric cancer, endometrial cancer, esophageal cancer, lung cancer, ovarian cancer, gastroesophageal junction cancer, breast cancer, bladder cancer, and renal cancer.

37. An antibody or an antigen-binding portion thereof, or a pharmaceutical combination for use according to any one of claims 12 to 14, wherein the patient is a human.

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