Combination of Faecalibacterium prausnitzii with anti-PD-1, anti-PD-L1 or anti-CTLA-4 antibodies for the treatment of cancer

Abstract

The present invention relates to a combination of the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under accession number I-4573 with at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies and anti-CTLA-4 monoclonal antibodies, for use in preventing and / or treating cancer in a mammalian patient. Such combinations are contemplated.

Description

[Technical field]

[0001] The present invention relates to the field of immunology, more particularly to a combination of the Faecalibacterium prausnitzii bacterial strain with an immune checkpoint inhibitor, in particular a combination of the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 with an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and / or an anti-CTLA-4 monoclonal antibody, and their use in human medicine.

[0002] In particular, the present invention relates to a combination of the Faecalibacterium prausnitzii bacterial strain with an immune checkpoint inhibitor, in particular the combination of the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 with an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and / or an anti-CTLA-4 monoclonal antibody, in particular an anti-PD-1 monoclonal antibody and / or an anti-PD-L1 monoclonal antibody. [Background technology]

[0003] Cancer is ranked as the first or second leading cause of death. In 2020, an estimated 19.3 million new cases of cancer were reported worldwide, and approximately 10 million deaths were attributed to cancer. The number of cancer patients worldwide is expected to reach 28.4 million in 2040, a 47% increase from 2020, with a larger increase in transitioning and transitioned countries due to demographic changes. In this field, there is often no or inadequate treatment. Thus, there is a well-recognized need to develop new and improved therapies to treat cancer.

[0004] Immunotherapy offers real hope in the treatment of many cancers, especially those that are not currently effectively cured by conventional therapy. This treatment mainly consists of the administration of monoclonal antibodies directed against tumor cells or against activators or inhibitors of checkpoints of the immune response against cancer (also called ICIs for immune checkpoint inhibitors). In particular, great progress has been made in the treatment of cancer by using ICIs that target cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), programmed cell death protein (PD-1), or programmed death-ligand 1 (PD-L1).

[0005] However, in most cases, the response rates remain relatively low. In fact, depending on the cancer considered, 40% to 95% of patients are resistant to ICI-based treatment. The mechanisms of escape are complex and are currently being actively studied by the scientific community. Summary of the Invention [Problem to be solved by the invention]

[0006] Thus, there remains a need in the art to provide effective and improved therapeutic approaches for treating cancer.

[0007] In particular, there is a need in the art to provide means for improving the efficacy of immune checkpoint inhibitors (ICIs), particularly to enhance their efficacy against tumors that are resistant to such treatment. [Means for solving the problem]

[0008] The present invention relates in particular to the following items: Item 1 : 1. A combination for use in preventing and / or treating cancer in a mammalian patient, comprising: the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under the accession number I-4573; at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; The above combinations.

[0009] In fact, we show for the first time that the in vivo administration of the bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under accession number I-4573, is able to restore the efficacy of immune checkpoint inhibitors against tumors in individuals.

[0010] In particular, the inventors have confirmed for the first time that alteration of the microbiota of tumor-bearing individuals by the administration of antibiotics impairs the antitumor effect of ICI treatment, in particular the antitumor effect of anti-PD-L1 treatment (Routy et al., Science 359, 91-97 (2018)) and serves as a suitable model for non-responders to mono-ICI therapy (i.e., individuals whose tumors are resistant to ICI-based treatment). Based on this, the inventors have surprisingly confirmed that the efficacy of anti-PDL-1 is significantly restored by administering to said individuals the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under the accession number I-4573.

[0011] Such a result is highly unexpected, taking into consideration that the Faecalibacterium prausnitzii bacterial strain specifically embodied in the present invention, i.e. the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573, was previously known for its anti-inflammatory properties (International Publication No. WO 2017 / 129515). Conversely, the present invention demonstrates for the first time the ability of this strain to enhance the immunostimulatory properties of immune checkpoint inhibitors in individuals in need of said immune checkpoint inhibitors.

[0012] Recent studies have demonstrated that the gut microbiome can modulate the response to anti-PD-1 immunotherapy in melanoma patients, and that fecal microbiota transplantation (FMT) using stool from responders to anti-PD-1 therapy was able to improve the response of mice administered anti-PD-L1 therapy (Gopalakrishnan et al. Science 359, 97-103 (2018); Davar et al., Science 371, 595-602 (2021) et Baruch et al. Science 10.1126 / science.abb5920 (2020)). None of these studies were able to determine the actual mechanism of action that led to this result, and in particular, which of the numerous bacteria present in the stool were responsible for this improved response, and which were pure biomarkers that would provide some insight into the likelihood that a given patient will respond to ICI treatment or not. Another problem associated with and identified by these studies is that the administration of stool, especially from responders to anti-PD-1 / anti-PD-L1 therapy, is not reproducible in its composition, especially the number and types of bacteria contained therein, which (i) vary from donor to donor and (ii) may even change over time within the same donor. Moreover, the shift in the patient's microbiota composition to that of the donor after fecal microbiota transplant may not be maintained over time, especially after antibiotic exposure.

[0013] Thus, the inventors have succeeded for the first time in identifying a new, effective and completely safe method for improving the efficacy of immune checkpoint inhibitors, in particular a means for enhancing their efficacy against tumors resistant to such treatment.

[0014] Item 2:2. The combination for use according to item 1, wherein the immune checkpoint inhibitor is an anti-PD-L1 monoclonal antibody or an anti-PD-1 monoclonal antibody, in particular an anti-PD-L1 monoclonal antibody.

[0015] Item 3: 3. The combination for use according to item 1 or 2, wherein the mammalian patient is selected from the group consisting of primates and humans.

[0016] Item 4: 4. A combination for use according to any one of items 1 to 3, wherein the mammalian patient is a human.

[0017] Item 5: 5. The combination for use according to any one of items 1 to 4, wherein the mammalian patient has intestinal microbial dysbiosis.

[0018] Item 6: The cancer may be a solid tumor or a liquid tumor, and may be, in particular, fibrosarcoma, bladder cancer, breast cancer, cervical cancer, colorectal cancer, kidney cancer, lung cancer, lymphoma, leukemia, myeloma, melanoma, neuroblastoma, Wilms' tumor, oral or oropharyngeal cancer, pancreatic cancer, prostate cancer, retinoblastoma, thyroid cancer, uterine cancer, adenoid cystic carcinoma, adrenal cortical tumor, chondrosarcoma, desmoid tumor, desmoplastic small round cell tumor, endocrine tumor, endodermal sinus tumor, epithelioid hemangioendothelioma, Ewing's sarcoma, nephroma, adrenal tumor, amyloidosis, anal cancer, appendix cancer, bile duct cancer, glioma, non-rhabdomyosarcoma soft tissue sarcoma (NRSTS). 6. The combination for use according to any one of items 1 to 5, wherein the cancer is selected from the group consisting of sarcoma, paraspinal sarcoma, renal cell carcinoma, rhabdomyosarcoma, synovial sarcoma, bone cancer, brain tumor, central nervous system tumor, cervical cancer, esophageal cancer, eye cancer, eyelid tumor, digestive tract cancer, HIV / AIDS-related cancer, lacrimal gland cancer, laryngeal or hypopharyngeal cancer, leukemia, liver cancer, meningioma, nasopharyngeal cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, parathyroid cancer, penile cancer, salivary gland cancer, sarcoma, non-melanoma skin cancer, small intestine cancer, gastric cancer, testicular cancer, thymoma and thymic cancer, vaginal cancer, and vulvar cancer.

[0019] Item 7: 7. The combination for use according to any one of items 1 to 6, wherein the cancer is fibrosarcoma.

[0020] Item 8: 8. The combination for use according to any one of items 1 to 7, wherein the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and the at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and an anti-CTLA-4 monoclonal antibody, are administered to the mammalian patient in the same composition or in separate compositions, preferably in separate compositions, wherein the one or more compositions further comprises a physiologically acceptable medium.

[0021] Item 9: 9. The combination for use according to any one of items 1 to 8, wherein the combination further comprises at least one additional anti-cancer agent which is distinct from the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and which is distinct from the anti-PD-1 monoclonal antibody, the anti-PD-L1 monoclonal antibody or the anti-CTLA-4 monoclonal antibody.

[0022] Item 10: The at least one additional anti-cancer agent is selected from the group consisting of monoclonal antibodies or bispecific antibodies, in particular Anti-CD137 monoclonal or bispecific antibody, anti-TIM-3 monoclonal or bispecific antibody, anti-B7-H3 monoclonal or bispecific antibody, anti-CD134 monoclonal or bispecific antibody, anti-CD154 monoclonal or bispecific antibody, anti-LAG-3 monoclonal or bispecific antibody, anti-CD227 monoclonal or bispecific antibody, anti-BTNA3 monoclonal or bispecific antibody, anti-CD39 monoclonal or bispecific antibody, anti-CD73 monoclonal or bispecific antibody, anti-CD115 monoclonal or bispecific antibodies, anti-SIRP alpha monoclonal or bispecific antibodies, anti-SIRP gamma monoclonal or bispecific antibodies, anti-CD28 monoclonal or bispecific antibodies, anti-NCR monoclonal or bispecific antibodies, anti-NKp46 monoclonal or bispecific antibodies, anti-NKp30 monoclonal or bispecific antibodies, anti-NKp44 monoclonal or bispecific antibodies, anti-NKG2D monoclonal or bispecific antibodies, and DNAM-1 monoclonal or bispecific antibodies; and selected from the group consisting of anti-CTLA-4 / anti-PDL-1 bispecific antibodies, anti-CTLA-4 / anti-PD-1 bispecific antibodies, and anti-PD-1 / anti-PD-L1 bispecific antibodies; Item 9. A combination for use according to item 9.

[0023] Item 11: the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under the accession number I-4573; at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; A combination of The combination, in particular, is one as described in any one of items 2, 3, 9 and 10. [Brief description of the drawings]

[0024] (Figure 1A) JPEG2024538781000001.jpg49170 (Figure 1B) JPEG2024538781000002.jpg52170 (Figure 1C) JPEG2024538781000003.jpg58170 (Figure 2) JPEG2024538781000004.jpg63170 (Figure 3) JPEG2024538781000005.jpg63170 (Figure 4) JPEG2024538781000006.jpg70170 (Figure 5A) JPEG2024538781000007.jpg47170 (Figure 5B) JPEG2024538781000008.jpg52170 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] definition

[0026] In order to provide a more complete understanding of the present invention, certain definitions are set forth below. Such definitions are intended to encompass all grammatical equivalents.

[0027] The term "antibody" is used herein in the broadest sense. An "antibody" refers to any polypeptide that contains at least (i) an Fc region and (ii) a binding polypeptide domain derived from a variable region of an immunoglobulin. Thus, antibodies include, but are not limited to, full length immunoglobulins, antibodies, antibody conjugates, and fragments of each. The terms "antibody" and "immunoglobulin" may be used interchangeably herein. More particularly contemplated antibodies herein are monoclonal antibodies, particularly immune checkpoint inhibitor (ICI) monoclonal antibodies, more particularly monoclonal antibodies selected from the group consisting of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies and anti-CTLA-4 monoclonal antibodies, particularly monoclonal antibodies selected from the group consisting of anti-PD-1 monoclonal antibodies and anti-PD-L1 monoclonal antibodies.

[0028] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous (i.e., the individual antibodies comprising the population are identical except for naturally occurring mutations that may be present in minor amounts) antibodies. Monoclonal antibodies are highly specific, being directed against a single antigen. Moreover, in contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier "monoclonal" should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). The "monoclonal antibodies" may also be isolated from phage antibody libraries using the techniques described in, for example, Clackson et al., Nature 352:624-628 (1991) or Marks et al., J. MoI Biol. 222:581-597 (1991).

[0029] The term "bispecific antibody" or "bispecific antibodies (BsAbs)" refers to a molecule that combines the antigen-binding sites of two antibodies in a single molecule. Thus, bispecific antibodies are capable of simultaneously binding to two different antigens. Methods for obtaining such antibodies are well known by those skilled in the art.

[0030] In the context of the present invention, the words "prevent" and "prevention" mean to reduce to a lower extent the risk or the probability of occurrence of a given phenomenon, i.e. in this invention cancer, in particular the cancers detailed hereinafter.

[0031] As used herein, the term "treat" or "treatment" includes alleviation and / or elimination of symptoms associated with a particular disorder or condition, i.e., in the present invention, cancer, particularly cancer as described in more detail below.

[0032] In the present invention, the term "immune checkpoint inhibitors" (also referred to as ICIs) refers to a type of immunomodulatory anticancer drug that targets immune checkpoints, which are key regulators of the immune system that, when stimulated, can dampen the immune response to immune stimulation. ICIs can be in various forms, such as antibodies (monoclonal, bispecific, etc.), RNAi molecules, or antisense molecules.

[0033] In the present invention, "intestinal microbial dysbiosis" is intended to designate a significant deviation from a balanced microbiota, either in terms of the overall microbiota profile, metabolism, or at the level of specific taxa.

[0034] "Physiologically acceptable medium" according to the present invention means a carrier or additive that is physiologically acceptable to the mammalian patient being treated while retaining the therapeutic properties of the compound being administered. One exemplary pharma- ceutically acceptable medium is physiological saline. Other physiologically acceptable mediums in the context of the present invention are known to those skilled in the art.

[0035] The terms "malignant cell," "cancer cell," and "tumor cell" may be used interchangeably herein. As used herein, a "tumor cell" refers to a cell that autonomously overgrows in vivo. Examples of tumor cells include (1) sarcomas, such as osteosarcoma and soft tissue sarcoma; (2) carcinomas, such as breast carcinoma, lung carcinoma, bladder carcinoma, thyroid carcinoma, prostate carcinoma, colon carcinoma, colorectal carcinoma, pancreatic carcinoma, gastric carcinoma, liver carcinoma, uterine carcinoma, cervical carcinoma, and ovarian carcinoma; (3) lymphomas, such as Hodgkin's lymphoma and non-Hodgkin's lymphoma; (4) neuroblastoma; (5) melanoma; (6) myeloma; (7) Wilms' tumor; and (8) leukemias, such as acute myelocytic leukemia (AML), chronic myelocytic leukemia (CML), acute lymphocytic leukemia (ALL), and / or other inflammatory diseases. leukemia, and chronic lymphocytic leukemia (CLL), (9) glioma, and (10) retinoblastoma.

[0036] As used herein, "cancer" refers to the uncontrolled abnormal proliferation of cells, and includes within its scope all known diseases caused by the uncontrolled abnormal proliferation of cells. Non-limiting examples of common cancers include fibrosarcoma, bladder cancer, breast cancer, ovarian cancer and stomach cancer, cervical cancer, colon cancer, endometrial cancer, head and neck cancer, lung cancer, melanoma, multiple myeloma, leukemia (e.g., myeloid leukemia, lymphocytic leukemia, myelocytic leukemia, and lymphoblastic leukemia), non-Hodgkin's lymphoma, prostate cancer, rectal cancer, malignant melanoma, especially fibrosarcoma.

[0037] The cancers considered herein may in particular be solid or liquid tumors.

[0038] In particular, the cancer cells of the present invention are selected from the group consisting of fibrosarcoma, bladder cancer, breast cancer, cervical cancer, colorectal cancer, kidney cancer, lung cancer, lymphoma, leukemia, myeloma, melanoma, neuroblastoma, Wilms' tumor, oral or oropharyngeal cancer, pancreatic cancer, prostate cancer, retinoblastoma, thyroid cancer, uterine cancer, adenoid cystic carcinoma, adrenal cortical tumor, chondrosarcoma, desmoid tumor, desmoplastic small round cell tumor, endocrine tumor, endodermal sinus tumor, epithelioid hemangioendothelioma, Ewing's sarcoma, nephroma, adrenal tumor, amyloidosis, anal cancer, appendix cancer, cholangiocarcinoma, glioma, non-rhabdomyosarcoma soft tissue sarcoma (NRSTS), and / or rhabdomyosarcoma soft tissue sarcoma (NRSTS). The cell may be contained in a cancer selected from the group consisting of sarcoma, paraspinal sarcoma, renal cell carcinoma, rhabdomyosarcoma, synovial sarcoma, bone cancer, brain tumor, central nervous system tumor, cervical cancer, esophageal cancer, eye cancer, eyelid tumor, gastrointestinal cancer, HIV / AIDS-related cancer, lacrimal gland cancer, laryngeal or hypopharyngeal cancer, leukemia, liver cancer, meningioma, nasopharyngeal cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, parathyroid cancer, penile cancer, salivary gland cancer, sarcoma, non-melanoma skin cancer, small intestine cancer, gastric cancer, testicular cancer, thymoma and thymic cancer, vaginal cancer, and vulvar cancer.

[0039] Combinations according to the invention

[0040] With a view to meeting the above mentioned objectives, and in particular to providing a new treatment for cancer in which the improved efficiency is not associated with any observable toxicity in the treated organism, the inventors have The bacterial strain Faecalibacterium prausnitzii, deposited at the Collection Nationale de Cultures de Microorganismes de l'Institut Pasteur de Paris (CNCM) on January 21, 2012 under the accession number I-4573, At least one immune checkpoint inhibitor We came up with a combination of the above.

[0041] The immune checkpoint inhibitor (ICI) according to the present invention may in particular be an antibody, in particular an anti-PD-1, anti-PD-L1 or anti-CTLA-4 antibody. More particularly, the at least one immune checkpoint inhibitor according to the present invention may be a monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and an anti-CTLA-4 monoclonal antibody.

[0042] The anti-PD-1 monoclonal antibody according to the present invention can be, for example, selected from the group consisting of Nivolumab, Pembrolizumab, and Cemiplimab.

[0043] The anti-PD-L1 monoclonal antibody according to the present invention may, for example, be selected from the group consisting of Atezolizumab, Avelumab, and Durvalumab.

[0044] The anti-CTLA-4 monoclonal antibody according to the present invention can be, for example, selected from the group consisting of ipilimumab, tremelimumab and quavonlimab.

[0045] In particular, the ICI of the combination according to the invention may be an anti-PD-L1 monoclonal antibody or an anti-PD-1 monoclonal antibody, in particular an anti-PD-L1 monoclonal antibody.

[0046] The Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 according to the combination of the present invention may be in isolated form. By "isolated form" according to the present invention is meant a form isolated from any fecal material or intestinal sampling. Thus, the bacterial strain of the present invention is not administered to the patient in the form of a fecal microbiota transplant (FMT). Methods for isolating bacteria from the intestinal microbiota are well known to those skilled in the art and are shown, for example, in Foditsch et al. (PLoS One. 2014; 9(12): e116465).

[0047] The mammalian patient according to the present invention may be selected from the group consisting of primates and humans, and may in particular be a human.

[0048] The mammalian patient according to the invention may in particular have an intestinal bacterial dysbiosis.

[0049] The combination according to the invention further comprises and which is different from the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under the accession number I-4573. Different from anti-PD-1 monoclonal antibody, anti-PD-L1 monoclonal antibody, or anti-CTLA-4 monoclonal antibody The composition may further comprise at least one additional anti-cancer agent.

[0050] The at least one additional anti-cancer agent may be selected from the group consisting of monoclonal or bispecific antibodies, in particular anti-CD137 monoclonal or bispecific antibodies, anti-TIM-3 monoclonal or bispecific antibodies, anti-B7-H3 monoclonal or bispecific antibodies, anti-CD134 monoclonal or bispecific antibodies, anti-CD154 monoclonal or bispecific antibodies, anti-LAG-3 monoclonal or bispecific antibodies, anti-CD227 monoclonal or bispecific antibodies, anti-BTNA3 monoclonal or bispecific antibodies, anti-CD39 monoclonal or bispecific antibodies, anti-CD7 3 monoclonal or bispecific antibodies, anti-CD115 monoclonal or bispecific antibodies, anti-SIRP alpha monoclonal or bispecific antibodies, anti-SIRP gamma monoclonal or bispecific antibodies, anti-CD28 monoclonal or bispecific antibodies, anti-NCR monoclonal or bispecific antibodies, anti-NKp46 monoclonal or bispecific antibodies, anti-NKp30 monoclonal or bispecific antibodies, anti-NKp44 monoclonal or bispecific antibodies, anti-NKG2D monoclonal or bispecific antibodies, and DNAM-1 monoclonal or bispecific antibodies; and Anti-CTLA-4 / anti-PDL-1 bispecific antibody, anti-CTLA-4 / anti-PD-1 bispecific antibody, and anti-PD-1 / anti-PD-L1 bispecific antibody may be selected from the group consisting of:

[0051] Medical Uses According to the Invention

[0052] As mentioned above, the present invention provides according to one of its aspects a combination for use in preventing and / or treating cancer in a mammalian patient, comprising: The Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under the accession number I-4573, more particularly said bacterial strain in isolated form, at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; The present invention relates to the above combination of

[0053] These ICIs, and in particular these monoclonal antibodies, are described throughout this specification.

[0054] To treat a patient in need thereof, such as those described above, a therapeutically effective dose of a combination according to the invention may be administered.

[0055] By "therapeutically effective dose" herein is meant a dose that produces the effect for which it is administered. The exact dose will depend on the purpose of the treatment and will be ascertainable by one skilled in the art using known techniques.

[0056] The dosage is, for example, 10 9 A daily dose equivalent to 10 colony forming units (CFU) 3 ~10 12 CFU, more preferably 10 7 ~l0 11 It could be CFU.

[0057] Dosages can also be used in pharmaceutical applications, e.g. 10 The daily dose is equivalent to 10 total cell count (TCC). 4 ~10 12 TCC, more specifically 10 8 ~ 12 It could be TCC.

[0058] Doses may also range from 0.001 to 100 mg per kg body weight (mg / kg) or greater of anti-PD-1 and / or anti-PD-L1 monoclonal antibodies according to the invention, for example, 0.1, 1.0, 5, 10 or 50 mg / kg of body weight.

[0059] The dosage and frequency of administration may be adapted depending on the host response as well as the frequency of injection for better tolerance.

[0060] For illustrative purposes only, the frequency of administration of the combination according to the invention can be daily administration for 5 to 15 consecutive days. Alternatively, the administration can be every 2, 3, 4, 5, 6 or 7 days for a period of up to several months. The Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 of the combination according to the invention and the immune checkpoint inhibitor (ICI) can be administered simultaneously or separately, for example separately during the same day or otherwise, for example with at least one or more days between the administration of the bacterial strain and the administration of the ICI.

[0061] As is known in the art, adjustments for protein degradation, systemic and local delivery, as well as age, weight, general health, sex, diet, time of administration, drug interactions, and severity of symptoms may be necessary and are readily determined by routine experimentation by one of ordinary skill in the art.

[0062] The administration of the combinations of the invention may be effected in various ways, in particular orally or rectally.

[0063] In certain embodiments, the conjugate according to the invention is in a form suitable for administration by rectal or oral route.

[0064] As described above, the present invention provides a combination for use in preventing and / or treating cancer in a mammalian subject, comprising: the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under the accession number I-4573; at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; The present invention relates to a combination of

[0065] The Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies and anti-CTLA-4 monoclonal antibodies, may be combined in the same composition or used in the form of separate compositions which may be administered simultaneously or sequentially.

[0066] In a particular embodiment, the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and the at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies and anti-CTLA-4 monoclonal antibodies, are administered in separate compositions.

[0067] When the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and an anti-CTLA-4 monoclonal antibody, are administered in separate compositions, the compositions can be administered to a mammalian patient via the same route or via different routes, simultaneously or separately.

[0068] By simultaneously, it is understood that the compositions can be administered at the same time, or on the same day or up to several days.

[0069] By separately it is understood that the compositions can be administered at least several days apart, for example at least 2 days apart.

[0070] In some embodiments, the composition comprising the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 according to the present invention and the ICI, in particular the anti-PD-1 monoclonal antibody, anti-PD-L1 monoclonal antibody and / or anti-CTLA-4 monoclonal antibody composition, is in liquid form.

[0071] In some embodiments, the composition comprising the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 according to the present invention and the ICI, in particular the anti-PD-1 monoclonal antibody, anti-PD-L1 monoclonal antibody and / or anti-CTLA-4 monoclonal antibody composition, is in a solid form, including a lyophilized form.

[0072] These compositions may be formulated according to standard methods, for example, as described in Remington: The Science and Practice of Pharmacy (Lippincott Williams & Wilkins; Twenty first Edition, 2005).

[0073] As previously indicated, the above-mentioned compositions contain physiologically / pharmaceutical acceptable vehicles / additives. The terms "physiologically acceptable" and "pharmaceutical acceptable" are used interchangeably herein.

[0074] The combinations of the present invention may be administered with other therapies or treatments, such as those mentioned above including antibiotics, cytotoxic chemotherapy, small molecules, other biologics, radiation therapy, surgery, etc., which are different from the additional anti-cancer agents mentioned above, and Unlike the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573, and It is different from anti-PD-1 antibodies, anti-PD-L1 antibodies, or anti-CTLA-4 antibodies. It is also different from.

[0075] According to a particular embodiment, the combination according to the invention may further comprise, as other therapeutic agent, at least one immunosuppressant, such as, for example, a glucocorticoid, a cytostatic drug (azathioprine, methotrexate), an antibody or a drug acting on an immunophilin (cyclosporine, tacrolimus, rapamycin).

[0076] These other therapeutic agents can be administered to a mammalian patient in the same composition or in separate compositions with the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 or with at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody and an anti-CTLA-4 monoclonal antibody.

[0077] In particular, these other therapeutic agents are administered in a composition separate from the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 or from at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of anti-PD-1 monoclonal antibodies, anti-PD-L1 monoclonal antibodies and anti-CTLA-4 monoclonal antibodies.

[0078] In a particular embodiment, the present invention provides a combination for the preparation of a medicament for preventing and / or treating cancer in a mammalian patient, comprising: The bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under the accession number I-4573, at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; Use of the above combinations.

[0079] In another embodiment, the present invention provides a method of preventing and / or treating cancer in a mammalian patient, the method comprising: The bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under the accession number I-4573, at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; to said mammalian patient a combination of

[0080] The present invention also relates to a combination for use as a medicament, comprising: The bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under the accession number I-4573, at least one immune checkpoint inhibitor, in particular at least one monoclonal antibody selected from the group consisting of an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, and an anti-CTLA-4 monoclonal antibody; The present invention relates to the above combination of

[0081] The invention is further illustrated by the following examples.

[0082] Working Example:

[0083] Cancer cell lines and tumor induction in mice

[0084] Sarcoma MCA-205 tumor cell line was cultured in vitro in DMEM supplemented with 10% FBS, 1% penicillin-streptomycin, and 1 mM HEPES according to the supplier's specifications.

[0085] Sarcoma MCA-205 tumor cells (500,000 cells / 100 μL / mouse) were inoculated subcutaneously into the right flank of immunocompetent C57Bl / 6J mice (9-week-old female).

[0086] Animal Group

[0087] The aim of this study was to evaluate the ability of the Faecalibacterium prausnitzii bacterial strain, deposited at the CNCM under accession number I-4573, to restore the antitumor effect of an anti-PD-L1 antibody in the MCA205 tumor-bearing mouse model with a dysregulated microbiota.

[0088] Microbiota dysbiosis was induced by administration of antibiotics (ABX).

[0089] Fifteen days before tumor inoculation, the mice were randomized according to body weight and assigned to ABX-treated and ABX-untreated groups.

[0090] MCA-205 sarcoma cells were inoculated subcutaneously into 9-week-old female C57BL / 6 mice (Charles River Laboratories) by injecting the cells into the flank on day 0. Five days after tumor inoculation, mice were randomized based on tumor volume and assigned to vehicle-treated and anti-PD-L1+ / - test bacteria-treated groups.

[0091] Treatments were administered as follows (10 mice per group): Group 1: No antibiotics (no ABX) and no treatment (vehicle: 200 μL PBS / glycerol, by oral gavage, once daily starting 6 days after tumor inoculation until the end of the study). Group 2: No ABX + anti-PD-L1 + vehicle treatment group: 200 μL PBS / glycerol, by oral gavage, once daily starting 6 days after tumor inoculation until the end of the study (day 42); and anti-PD-L1 antibody, 5 mg / kg, intraperitoneally, once daily on days 6, 9, 12 and 15. Group 3: treatment with antibiotics (ABX) and no treatment (vehicle: antibiotic combination in drinking water, starting 14 days before tumor inoculation until the end of the study; and 200 μL PBS / glycerol, by oral gavage, once daily, starting 6 days after tumor inoculation until the end of the study). Group 4: Antibiotic (ABX) treatment + anti-PD-L1 + vehicle treatment group: Antibiotic combination in drinking water starting 14 days before tumor inoculation until the end of the study; anti-PD-L1 antibody, 5 mg / Kg, intraperitoneally, once daily on days 6, 9, 12 and 15; and 200 μL PBS / glycerol by oral gavage, once daily starting 6 days after tumor inoculation until the end of the study. Group 5: treatment with antibiotics (ABX) + anti-PD-L1 + treatment with Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 group: drinking water solution combined with antibiotics, starting 14 days before tumor inoculation until the end of the study; anti-PD-L1 antibody, 5 mg / Kg, intraperitoneally, once daily, on days 6, 9, 12 and 15; and 200 μL (1x10 10 (TCC) strain CNCM I-4573, administered by oral gavage once daily starting 6 days after tumor inoculation until the termination of the study.

[0092] Starting 5 days after tumor cell inoculation, all experimental animal groups, each consisting of 10 mice, were monitored three times a week up to day 30 for the following parameters: Tumor size: measured by physical examination three times a week Body weight: monitored once a week before tumor inoculation and three times a week thereafter Survival rates: 3x per week, presented as Kaplan-Meier plots.

[0093] Antibiotic Treatment (ABX)

[0094] Antibiotic solutions consisting of ampicillin (Sigma - 1 mg / mL), streptomycin (Sigma - 5 mg / mL) and colistin (Carbosynth - 1 mg / mL) were prepared weekly in drinking water supplemented with 1% sucrose (Sigma) and kept refrigerated. Cage bottles were changed once a week.

[0095] Tested bacteria of the present invention

[0096] The bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under the accession number I-4573, was prepared ready to use in PBS / glycerol. Treatments were started on day 6 after tumor inoculation and were maintained throughout the study period, with the treatments being performed once a day (including weekends).

[0097] Treatment was performed by oral gavage of 200 μL per mouse per day, which was 10 10 This is equivalent to TCC.

[0098] anti-PD-L1

[0099] Anti-PD-L1 treatment was administered by intraperitoneally administering 100μg (for a 20g mouse) of anti-PD-L1 antibody (5mg / Kg). Treatment was repeated four times on days 6, 9, 12 and 15 after tumor cell inoculation. Anti-PD-L1 antibody (clone 10F.9G2-BioXCell) was diluted to a concentration of 1mg / mL in sterile PBS. The stock solution was then aliquoted (amount for one day's treatment) and stored at 4℃.

[0100] Example 1: Effect of ABX on anti-PD-L1 mediated responses

[0101] The survival rates over 30 days of MCA-205 tumor-bearing mice according to groups 1, 2, 3 and 4 were first examined.

[0102] The results obtained are presented in FIG. 1A.

[0103] After 30 days, 90% of mice in group 2 were alive, compared with only 40% in group 4, demonstrating that antibiotic-induced changes in the microbiota compromised the antitumor effect of anti-PD-L1 treatment.

[0104] This is also shown in Figures 1B and 1C, where tumor volumes were measured in mice from the same four groups.

[0105] The mean tumor volume of mice from group 4 (ABX + anti-PD-L1) was significantly higher than that of mice from group 2 (anti-PD-L1).

[0106] The results also demonstrate the detrimental impact of antibiotic-induced microbiota alteration on the antitumor properties of anti-PD-L1 treatment.

[0107] Example 2: Effect of bacterial strains according to the invention on ABX-mediated responses

[0108] The changes in tumor volume over 26 days in MCA-205 tumor-bearing mice from groups 2, 4 and 5 were also examined.

[0109] The results obtained are presented in FIGS.

[0110] After 26 days, the average tumor volume in mice in group 4 was 1000 mm 3 whereas the corresponding values ​​in group 5 were statistically inferior, at only about 500 mm 3 , similar to that observed in mice from group 2.

[0111] The survival progression of the tested mice was also measured during the experiment.

[0112] The results obtained are presented in FIG.

[0113] After 30 days, only 40% of the individuals from group 4 treated with antibiotics (ABX + anti-PD-L1) were alive. In contrast, 90% of the individuals in group 5 were still alive after 30 days due to administration of the bacterial strain according to the invention. This percentage is the same as that obtained for group 2 in Figure 1A.

[0114] When tumor-bearing mice with microbiota dysbiosis are treated with a bacterial strain according to the invention in combination with PD-L1, survival progression is significantly improved compared to anti-PD-L1 alone.

[0115] These results demonstrate the ability of the bacterial strains according to the invention to significantly improve the anti-tumor effect of anti-PD-L1 treatment.

[0116] These results therefore substantiate the fact that the bacterial strain according to the invention has the potential to rescue the antitumor effect of ICI in non-responders.

[0117] Example 3: Complementary study of the antitumor effect of the bacterial strain according to the invention on the MCA205 tumor-bearing mouse model with a dysregulated microbiota

[0118] The same protocol as detailed above is carried out on the following two groups of MCA205 tumor-bearing mice (10 mice per group): Group 3' (same as Group 3 above): treatment with antibiotics (ABX) and no treatment (vehicle: antibiotic combination drinking water solution, starting 14 days before tumor inoculation until the end of the study; and 200 μL PBS / glycerol, by oral gavage, once daily, starting 6 days after tumor inoculation until the end of the study). Group 6: treatment with antibiotics (ABX) + treatment with the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under the accession number I-4573: drinking water solution combined with antibiotics, starting 14 days before tumor inoculation until the end of the study (D42); 10 (TCC) CNCM I-4573 strain, administered by oral gavage once daily starting 6 days after tumor inoculation until the end of the study (D42).

[0119] Starting 5 days after tumor cell inoculation, all experimental animal groups, each consisting of 10 mice, were monitored three times a week until day 42 for the following parameters: Tumor size: measured by physical examination three times a week Body weight: monitored once a week before tumor inoculation and three times a week thereafter Survival rates: 3x per week, presented as Kaplan-Meier plots.

[0120] Antibiotic solutions were identical to those described above and consisted of ampicillin (Sigma - 1 mg / mL), streptomycin (Sigma - 5 mg / mL), and colistin (Carbosynth - 1 mg / mL), prepared weekly in drinking water supplemented with 1% sucrose (Sigma) and kept refrigerated. Cage bottles were changed weekly. Treatment was initiated 14 days prior to tumor inoculation and continued throughout the study.

[0121] The bacterial strain Faecalibacterium prausnitzii, deposited at the CNCM under accession number I-4573, was prepared ready for use as previously described, i.e. in PBS / glycerol. Treatments were started on day 6 after tumor inoculation and were maintained throughout the study period, with the treatments being performed once a day (including weekends).

[0122] Treatment was performed by oral gavage of 200 μL per mouse per day, which was 10 10 This is equivalent to TCC.

[0123] result

[0124] The mean tumor volume and survival rate over 26 days of MCA-205 tumor-bearing mice were examined according to group 3' (ABX+vehicle) and group 6 (ABX+CNCM I-4573), and the results are shown in Figure 5A and Figure 5B, respectively.

[0125] At the end of the experiment, the data show that when mice were treated with ABX, no advantage was observed with CNCM I-4573 alone in either mean tumor volume progression or survival proportion.

[0126] JPEG2024538781000009.jpg255165

Claims

1. 1. A combination for use in preventing and / or treating cancer in a mammalian patient, comprising: the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under accession number I-4573; and At least one immune checkpoint inhibitor The above combination.

2. 2. The combination for use according to claim 1, wherein the immune checkpoint inhibitor is an anti-PD-L1 monoclonal antibody or an anti-PD-1 monoclonal antibody.

3. 2. The combination for use according to claim 1, wherein said mammalian subject is selected from the group consisting of primates and humans.

4. 2. The combination for use according to claim 1, wherein the mammalian patient is a human.

5. 2. The combination for use according to claim 1, wherein the mammalian patient has intestinal bacterial dysbiosis.

6. The cancer may be a solid tumor or a liquid tumor, in particular fibrosarcoma, bladder cancer, breast cancer, cervical cancer, colorectal cancer, kidney cancer, lung cancer, lymphoma, leukemia, myeloma, melanoma, neuroblastoma, Wilms' tumor, oral or oropharyngeal cancer, pancreatic cancer, prostate cancer, retinoblastoma, thyroid cancer, endometrial cancer, adenoid cystic carcinoma, adrenal cortical tumor, chondrosarcoma, desmoid tumor, desmoplastic small round cell tumor, endocrine tumor, endodermal sinus tumor, epithelioid hemangioendothelioma, Ewing's sarcoma, nephroma, adrenal tumor, amyloidosis, anal cancer, appendix cancer, bile duct cancer, glioma ...

2. The combination for use according to claim 1, wherein the cancer is selected from the group consisting of: rhabdomyosarcoma, non-rhabdomyosarcoma soft tissue sarcomas (NRSTS), paraspinal sarcoma, renal cell carcinoma, rhabdomyosarcoma, synovial sarcoma, bone cancer, brain tumor, central nervous system tumor, cervical cancer, esophageal cancer, eye cancer, eyelid tumor, gastrointestinal cancer, HIV / AIDS-related cancer, lacrimal gland cancer, laryngeal or hypopharyngeal cancer, leukemia, liver cancer, meningioma, nasopharyngeal cancer, ovarian cancer, fallopian tube cancer, peritoneal cancer, parathyroid cancer, penile cancer, salivary gland cancer, sarcoma, non-melanoma skin cancer, small intestine cancer, gastric cancer, testicular cancer, thymoma and thymic cancer, vaginal cancer, and vulvar cancer.

7. 2. The combination for use according to claim 1, wherein the cancer is fibrosarcoma.

8. The combination for use according to claim 1, wherein the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and the at least one immune checkpoint inhibitor are administered to the mammalian patient in the same composition or in separate compositions, wherein the one or more compositions further comprise a physiologically acceptable medium.

9. 2. The combination for use according to claim 1, wherein the combination further comprises at least one additional anti-cancer agent which is different from the Faecalibacterium prausnitzii bacterial strain deposited at the CNCM under accession number I-4573 and which is different from an anti-PD-1 monoclonal antibody, an anti-PD-L1 monoclonal antibody, or an anti-CTLA-4 monoclonal antibody.

10. The at least one additional anti-cancer agent is selected from the group consisting of a monoclonal antibody or a bispecific antibody, in particular Anti-CD137 monoclonal or bispecific antibody, anti-TIM-3 monoclonal or bispecific antibody, anti-B7-H3 monoclonal or bispecific antibody, anti-CD134 monoclonal or bispecific antibody, anti-CD154 monoclonal or bispecific antibody, anti-LAG-3 monoclonal or bispecific antibody, anti-CD227 monoclonal or bispecific antibody, anti-BTNA3 monoclonal or bispecific antibody, anti-CD39 monoclonal or bispecific antibody, anti-CD73 monoclonal or bispecific antibody, anti-CD115 Monoclonal or bispecific antibodies, anti-SIRP alpha monoclonal or bispecific antibodies, anti-SIRP gamma monoclonal or bispecific antibodies, anti-CD28 monoclonal or bispecific antibodies, anti-NCR monoclonal or bispecific antibodies, anti-NKp46 monoclonal or bispecific antibodies, anti-NKp30 monoclonal or bispecific antibodies, anti-NKp44 monoclonal or bispecific antibodies, anti-NKG2D monoclonal or bispecific antibodies, and DNAM-1 monoclonal or bispecific antibodies; and Anti-CTLA-4 / anti-PDL-1 bispecific antibody, anti-CTLA-4 / anti-PD-1 bispecific antibody, and anti-PD-1 / anti-PD-L1 bispecific antibody selected from the group consisting of A combination for use according to claim 9.

11. the bacterial strain Faecalibacterium prausnitzii deposited at the CNCM under accession number I-4573; and At least one immune checkpoint inhibitor A combination of The combination, in particular as defined in claims 2, 3, 9 and 10.

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