A cancer vaccine obtained by co-culturing and immunizing autologous and / or haploidentical natural killer (NK) cells with autologous cancer cells attenuated under far-UVC light

Abstract

The invention relates to a personalized cancer vaccine obtained by co-culturing and immunization of autologous and / or haploidentical natural killer (NK) cells with autologous cancer cells attenuated under Far-UVC (Ultraviolet) light, in the field of cancer immunotherapy.

Description

[0001] A CANCER VACCINE OBTAINED BY CO-CULTURING AND IMMUNIZING AUTOLOGOUS AND / OR HAPLOIDENTICAL NATURAL KILLER (NK) CELLS WITH AUTOLOGOUS CANCER CELLS ATTENUATED UNDER FAR-UVC LIGHT

[0002] Technical Field

[0003] The invention relates to a personalized cancer vaccine obtained by co-culturing and immunization of autologous and / or haploidentical natural killer (NK) cells with autologous cancer cells attenuated under Far-UVC (Ultraviolet) light, in the field of cancer immunotherapy.

[0004] Background of the Invention

[0005] Cancer is characterized by the uncontrolled proliferation of body cells. Potential cancers, largely initiated by mutations affecting the cell regulatory function in the genome, constantly arise in the body.

[0006] However, natural processes such as apoptosis and immune surveillance eliminate these cancers before tumors develop. Despite the protection it provides, immune surveillance directs the selection of malignancies. This process, called immune regulation, explains the mechanism by which cancers arise in individuals with competent immune systems. In short, malignant cells recognized by the immune system are destroyed, while malignant cells that are not recognized survive.

[0007] This repeated selection of immunologically silent cancerous cells eventually leads to the formation of a tumor mass. This tumor microenvironment is immunosuppressive due to the secretion of inhibitory cytokines, the expression of inhibitory membrane-bound ligands, and the accumulation of regulatory cell populations that deactivate tumor-specific immune cells. These factors create an environment that supports cancer growth, unhindered by the body's natural defenses.

[0008] Small molecule inhibitors, as well as biological agents designed against dri ver / actionable mutations, are designed against a single target at a time, requiring a long development process and resulting in a limited number of available mechanisms of action along with a number of associated side effects. The feasibility of a multivalent target therapy consisting of small molecules or biological agents is limited due to practical reasons and the cumulative side effects associated with therapeutic drugs.Although cancer treatment has made significant progress in the last decade, long-term responses are not achieved in most cases in patients with metastatic disease.

[0009] Standard treatments using surgical or radio / chemotherapy procedures for widespread tumors provide limited results that rarely alter the outcome of the disease. Over the last two decades, various immunotherapy approaches have been tested as alternative treatments against solid and hematological malignancies. The use of recombinant cytokines such as IL-2 and IFN-a, T cell-mediated adaptive therapies, monoclonal antibodies, and dendritic cell-based vaccines has resulted in improvements in tumor growth control and patient survival, among others. However, despite the relative effectiveness of these treatments, disadvantages remain prevalent in a large proportion of cases in terms of the percentage of resistant patients and side effects.

[0010] Intratumoral heterogeneity underlies the tumor variants that arise under targeted therapies and immunosuppression, and current treatments fail to provide a solution in cases of broad tumor heterogeneity.

[0011] While current cancer treatments such as chemotherapeutic agents and ionizing radiation reduce the recurrence rate in some cancer types, the resulting clones may be more resistant to the same treatments. Furthermore, these treatments are associated with a lack of specificity in the case of chemotherapy or a loss of hematopoietic potential following radiation therapy. While these problems remain unresolved, activation of the immune system against cancer remains a viable treatment option.

[0012] Consequently, a range of immunotherapeutic treatments have been investigated to enhance a patient's natural anti-tumor immune response and overcome tumor-induced immunosuppression. These treatments, varying widely in composition, aim to induce specific anti-tumor immunity.

[0013] Allogeneic vaccines are largely similar to autologous vaccines, except that the material is derived from anothermember of the same species. Commonly used allogeneic materials include the use of established cancer cell lines grown in the laboratory that are known to express TAAs of a specific tumor type. This allows for mass production, storage, and modification of these therapeutics before use. Although many allogeneic cell-based vaccines are derived from cancer cell lines of the same species and type (breast, prostate, lung, etc.), they may differ from autologous vaccines in that they do not contain “patient-specific” tumor antigens. However, the greatest obstacle to the use of allogeneic therapies is the widespread intratumoral heterogeneity in cancers.The use of allogeneic whole tumor cells for vaccination has several advantages, as they are immortal cell lines with simple and standardized production protocols, making them cost-effective sources of good tumor antigens and allowing for comparison of clinical responses among patients. Vaccination with autologous tumor cells has many benefits because they are a large source of tumor antigens. However, the effectiveness of the vaccine is limited if sufficient antigens are not present in the tumor.

[0014] A personalized cancer vaccine needs to target a large number of tumor-specific mutations, minimizing side effects while protecting normal tissue and keeping tumors under immunological memory control for as long as possible, but current treatments cannot achieve this.

[0015] Tumor cell lysates have also been used as vaccines to stimulate anti-tumor immune responses. While these vaccines allow the presentation of multiple tumor antigens, they exhibit poor immunogenicity, and some have been shown to be completely ineffective.

[0016] The inability of tumor cell lysates to induce a sustained immune response may be partly due to the presence of immunosuppressive molecules within the lysate. Patients often have severely weakened immune systems due to prior participation in various chemotherapy and radiation treatments.

[0017] Neoantigens are important targets for immune responses. Current cancer vaccine therapies are limited to neoantigens, and therefore immune responses are often neoantigen-specific.

[0018] Clinical trials and regulatory procedures for oral vaccines may result in lower costs due to less complex application processes. Injectable vaccines typically require more complex manufacturing processes, including purification and formulation of vaccine antigens. Manufacturing costs are higher, and animal and clinical trials are more complicated and costly. Cytotoxic T cells induced via subcutaneous vaccination with a peptide-based cancer vaccine are delivered to the vaccination site but do not accumulate within the tumor, resulting in limited cancer regression. A vaccination method containing more heterogeneous antigens and exposure to the tumor from a larger surface area rather than a limited area such as subcutaneous area is needed for a stronger response.

[0019] In vaccine therapies with allogeneic tumor cells, the mismatched minor histocompatibility antigen poses an additional risk consistent with the increased graft-versus-tumor responses observed in patients receiving bone marrow transplants. Allogeneic tumor cell vaccines do not include strategies to mitigate tumor-induced immunosuppression.

[0020] Current cancer treatments generally rely on methods such as chemotherapy, radiotherapy and immunotherapy, the effectiveness of which varies from patient to patient, and there iscurrently no definitively successful cancer treatment method. Existing immunotherapeutic approaches developed to enable the immune system to generate a specific response against cancer have limited success rates. There is a need for a new generation of therapeutic cancer vaccines that can generate a stronger and more specific immune response against individual cancer cells while reducing systemic toxicity.

[0021] In the literature, the European patent document numbered EP3892295B1 refers to this subject as follows: “The present invention relates to the provision of vaccines that are specific to a patient’s tumor and potentially useful for immunotherapy of both the primary tumor and tumor metastases. In one aspect, the present invention relates to a method involving the following steps to provide a personalized cancer vaccine: (a) identifying cancer-specific somatic mutations in a tumor sample of a cancer patient to provide a cancer mutation signature of the patient and (b) providing a vaccine exhibiting the cancer mutation signature characteristic obtained in step (a). In an additional aspect, the present invention relates to vaccines that can be obtained by said method.”

[0022] Said application describes the provision of vaccines that are specific to a patient's tumor and potentially useful for immunotherapy of both the primary tumor and tumor metastases.

[0023] Furthermore, in the literature, the European patent document numbered EP3473267B1 refers to this subject as follows: “The present invention relates to the provision of vaccines that are tumorspecific and potentially useful for immunotherapy of both the primary tumor and tumor metastases. In one aspect, the present invention relates to a method involving the following steps to provide a personalized cancer vaccine: (a) identifying cancer-specific somatic mutations in a tumor sample of a cancer patient to provide a cancer mutation signature of the patient, and (b) providing a vaccine exhibiting the cancer mutation signature characteristic obtained in step (a). In an additional aspect, the present invention relates to vaccines that can be obtained by said method.”

[0024] Said patent also describes the provision of vaccines that are tumor-specific and potentially useful for immunotherapy of both the primary tumor and tumor metastases.

[0025] Similarly, in the literature, the European patent document numbered EP2184072B1 refers to this subject as follows: “The present invention relates to biotechnology, particularly human health. In an embodiment, the present invention describes vaccine compositions for protective and / or therapeutic cancer vaccines, primarily aimed at generating a significant increase in the immune response against Epidermal Growth Factor (EGF), which has been extensively demonstrated for the growth of tumors of epithelial origin and of oncological importance.”Said patent describes the production of a homogeneous vaccine formulation for cancer treatment.

[0026] For the reasons mentioned above, there is a need for a new, personalized cancer vaccine.

[0027] Objects of the Invention

[0028] Based on the current state of the art, the object of this invention is to develop a novel, personalized cancer vaccine.

[0029] Another object of the invention is to train natural killer (NK) cells obtained from the patient's own immune system and / or haploidentical NK cells obtained from parents, siblings, etc., by culturing them in vitro with cancer cells using a specific protocol, thereby creating a patientspecific, enhanced immunotherapeutic vaccine.

[0030] Another object of the invention is to create a structure that enables a stronger and more specific immune response against cancer by immunizing NK cells co-cultured with autologous cancer cells attenuated using Far-UVC light.

[0031] Detailed Description of the Invention

[0032] This present detailed description explains the novelty of the invention with examples that do not impose any limiting effect on a better understanding of the subject.

[0033] The invention relates to a method for the production of a personalized cancer vaccine, characterized in that it comprises the steps of isolating cancer cells from the patient via surgery or biopsy, proliferating the obtained cancer cells under suitable culture conditions, attenuating (weakening) the cancer cells with Far-UVC light to render them immunogenic but without proliferative capacity, isolating and activating autologous and / or alloidentical natural killer (NK) cells, training the natural killer (NK) cells using co-culture, and returning the prepared natural killer (NK) cells to the patient.

[0034] In the first step in the application of the invention:

[0035] - Autologous cancer cells are obtained from the patient. Cancer cells are isolated from the patient via surgery or biopsy, and the obtained cancer cells are multiplied under appropriate culture conditions.

[0036] - Cancer cells are attenuated (weakened) using Far-UVC light. In this process, autologous cancer cells multiplied in culture are exposed to Far-UVC light at a wavelength of 222 nm for a specific dose and duration. As a result of this process, the cells become immunogenic but have lost their proliferative capacity (attenuated).- Autologous and / or alloidentical natural killer (NK) cells are isolated and activated. Natural killer (NK) cells are isolated from the peripheral blood or bone marrow of the patient and / or their parents, siblings, etc. NK cells are activated with interleukins (such as IL-2, IL- 15) and other appropriate growth factors.

[0037] - Natural killer (NK) cells are trained using co-culture. Attenuated (weakened) autologous cancer cells are co-cultured in vitro with activated natural killer (NK) cells for a specific period. During this process, the natural killer (NK) cells are programmed to learn to recognize the surface antigens of cancer cells, thus generating a more specific anti-tumor response.

[0038] - The prepared natural killer (NK) cells are then returned to the patient. The trained natural killer (NK) cells, obtained from culture and having developed a specific immune response, are returned to the patient intravenously, intrathecally, via direct open surgery, or under ultrasound, MRI or CT guidance, or stereotactically into intracavitary site (into the tumor bed). This process enables the immune system to generate a stronger response against the targeted cancer cells, thus demonstrating a therapeutic effect.

[0039] The personalized cancer vaccine in question provides a personalized and specific immune response. Natural killer (NK) cells, trained with antigens obtained from the patient's own cancer cells, create an individualized and specific immune response.

[0040] Said cancer vaccine reduces side effects. Systemic toxicity is significantly reduced compared to traditional methods such as chemotherapy or radiotherapy.

[0041] The training and increased activation of these natural killer (NK) cells strengthens the innate immune response against cancer cells.

[0042] Trained natural killer (NK) cells are more effective at recognizing cancer cells, potentially reducing the risk of disease recurrence and providing a definitive cure.

[0043] This invention represents an innovative approach to developing immunotherapeutic cancer vaccines and can be implemented by pharmaceutical companies and research centers operating in the fields of biotechnology and oncology. This method, within the scope of personalized cancer treatments, can be used in advanced cellular therapy laboratories and clinical immunotherapy units.

[0044] The method involves developing an immunotherapeutic vaccine by co-culturing natural killer (NK) cells with autologous cancer cells attenuated (weakened) using Far-UVC light and returning them to the patient. This approach aims to provide a more effective cancer treatment by creating a personalized immune response.

Claims

CLAIMS1. A method for producing personalized cancer vaccines, characterized in that it comprises the steps of isolating cancer cells, proliferating the obtained cancer cells under suitable culture conditions, attenuating the cancer cells using Far-UVC light at a wavelength of 222 nm to render them immunogenic but devoid of proliferative capacity, isolating and activating autologous and / or alloidentical natural killer cells, training the natural killer cells using co-culture, and obtaining the prepared natural killer cells.

2. A method for producing personalized cancer vaccines according to any one of the preceding claims, characterized in that it comprises the step of isolating natural killer (NK) cells from the peripheral blood or bone marrow of the patient and / or their parents, siblings, etc.

3. A method for producing personalized cancer vaccines according to any one of the preceding claims, characterized in that it comprises the step of activating said natural killer (NK) cells using interleukins (IL-2, IL- 15) and other appropriate growth factors.

4. A method for producing personalized cancer vaccines according to any one of the preceding claims, characterized in that it comprises the step of co-culturing attenuated (weakened) autologous cancer cells with activated natural killer (NK) cells in an in vitro environment.

5. A method for producing personalized cancer vaccines according to any one of the preceding claims, characterized in that it comprises the step of programming said natural killer (NK) cells to learn to recognize surface antigens of cancer cells, thereby generating a more specific anti-tumor response.

6. A method for producing personalized cancer vaccines according to any one of the preceding claims, characterized in that it comprises the step of reintroducing natural killer (NK) cells — which have been cultured and have developed a specific immune response — into the patient via intravenous, intrathecal, or direct open surgical routes, or intracavitary (into the tumor bed) under ultrasound, MRI, or CT guidance, or using stereotactic techniques.