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EP-4739322-A1 - PHARMACEUTICAL PRODUCT FOR AUTOLOGOUS ADOPTIVE CELLULAR THERAPY

EP4739322A1EP 4739322 A1EP4739322 A1EP 4739322A1EP-4739322-A1

Abstract

The present invention concerns a pharmaceutical product for use as an adoptive autologous cellular therapy against a disease in a human or animal subject, comprising two compositions comprising various combinations of autologous immune cells to be administered in three separate doses sequentially to the human or animal subject.

Inventors

  • PAPASOTIRIOU, Ioannis

Assignees

  • R.G.C.C. Holdings AG

Dates

Publication Date
20260513
Application Date
20240705

Claims (19)

  1. 1. Pharmaceutical product for use in an autologous adoptive cellular therapy for the treatment of a cancer disease of a human or animal subject, comprising - a first dose comprising a first composition, comprising autologous macrophages derived from autologous mononuclear cells, and further comprising autologous natural killer cells, - a second dose comprising a second composition, and - a third dose comprising the second composition, wherein the second composition comprises ex vivo activated autologous dendritic cells, ex vivo activated autologous CD8+ cytotoxic T-lymphocytes and ex vivo activated autologous plasma cells.
  2. 2. Pharmaceutical product according to claim 1 , wherein in the first composition the macrophages are M1 -polarized autologous macrophages.
  3. 3. Pharmaceutical product according to claim 1 or 2, wherein in the first composition the autologous natural killer cells are autologous CD16+ and CD56+ natural killer cells.
  4. 4. Pharmaceutical product according to one of the preceding claims, wherein in the second composition contained in the second dose and in the third dose, the autologous dendritic cells are activated against one or more antigenic peptides, and wherein the autologous CD8+ cytotoxic T-lymphocytes are activated against one or more antigenic peptides, such that the activated autologous CD8+ cytotoxic T- lymphocytes are capable of recognizing the one or more antigenic peptides, and wherein the autologous plasma cells are activated against one or more antigenic peptides, such that the autologous plasma cells are capable of producing antibodies against the one or more antigenic peptides, wherein the antigenic peptides are isolated from the same human or animal subject.
  5. 5. Pharmaceutical product according to claim 4, wherein the activated autologous CD8+ cytotoxic T-lymphocytes are capable of recognizing at least one of the same antigenic peptides against which the autologous dendritic cells are activated, and/or wherein the activated autologous plasma cells are capable of producing antibodies against at least one of the same antigenic peptides against which the autologous dendritic cells are activated.
  6. 6. Pharmaceutical product according to one of the preceding claims, wherein the three doses are adapted for administration to the human or animal subject separately from each other and sequentially at three different points in time, wherein preferably the first dose comprising the first composition is adapted to be administered to the human or animal subject in week 1 of a treatment schedule, preferably by injection, wherein preferably the second dose comprising the second composition is adapted to be administered to the human or animal subject in week 3 of the treatment schedule, preferably by injection, and wherein preferably the third dose comprising the second composition is adapted to be administered to the human or animal subject in week 5 of the treatment schedule, preferably by injection.
  7. 7. Pharmaceutical product according to one of the preceding claims, wherein each of the first, second and third dose is adapted to be administered to the human or animal subject by intravenous injection.
  8. 8. Pharmaceutical product according to one of the preceding claims, for use as a medicament against a cancer disease of a human or animal subject.
  9. 9. Kit-of-parts for use as a medicament against a disease in a human or animal subject, preferably against a cancer disease, comprising the pharmaceutical product according to one of claims 1-7.
  10. 10. Kit-of-parts for use as a medicament against a cancer disease in a human or animal subject, comprising: - a first dose comprising a first composition, comprising macrophages derived from autologous mononuclear cells, preferably autologous macrophages polarized in a M1 -phase, and further comprising autologous natural killer cells, preferably CD16+/CD56+ natural killer cells, - a second dose comprising a second composition, and - a third dose comprising the second composition, wherein the second composition comprises ex vivo activated autologous dendritic cells, ex vivo activated autologous cytotoxic T-lymphocytes and ex vivo activated autologous plasma cells; and wherein preferably the first dose, the second dose and the third dose each are adapted for administration to the human or animal patient at a different point in time of a treatment schedule, preferably in 2-week intervals.
  11. 11. Method for the production of a medicament for autologous adoptive cellular therapy in the treatment of a cancer disease of a human or animal subject, comprising the following steps ex vivo: a.) providing a population of autologous macrophages, preferably autologous macrophages polarized in a M1 -phase; b.) providing a population of autologous natural killer cells, preferably CD16+/CD56+ natural killer cells; c.) providing a population of ex vivo activated autologous dendritic cells; d.) providing a population of autologous CD8+ cytotoxic T-lymphocytes; e.) providing a population of autologous plasma cells; f.) mixing of the population resulting from step a.) with the population resulting from step b.) for the provision of a first composition for administration in a first dose to a human or animal subject.; g.) mixing of a first portion of the population resulting from step c.) with the population from step d.), resulting in a population of ex vivo activated CD8+ cytotoxic T- lymphocytes; h.) mixing of a second portion of the population resulting from step c.) with the population resulting from step e.), resulting in a population of ex vivo activated autologous plasma cells; i.) mixing of the population resulting from step g.) with the population of step h.) for the provision of a second composition; j.) splitting up the second composition resulting from step i.) into two preferably equal portions, resulting in a second dose of the second composition and a third dose of the second composition for administration to the human or animal subject.
  12. 12. Method according to claim 11 , wherein in step g.), for the purpose of providing a population of activated autologous CD8+ cytotoxic T-lymphocytes, the population resulting from step c.) is incubated in the presence of p2-microglobulin, preferably for 6-18 hours, preferably for 8-12 hours, and preferably also incubated with at least one, preferably all of the following group consisting of IL-1 p, TNF-a, IL-6, PGE2 prior to mixing the population resulting from step c.) with the autologous CD8+ cytotoxic T-lymphocytes, preferably for 24-72 hours, more preferably for 48 hours.
  13. 13. Method according to claim 11 or 12, wherein in step g.), for the purpose of providing a population of ex vivo activated autologous CD8+ cytotoxic T-lymphocytes, the population resulting from step c.), after mixing with the population resulting from step d.), is further incubated with at least one of the cytokines of the group consisting of IL-2, IL-7, and IL-15, preferably for 8-15 days, more preferably for 10 days.
  14. 14. Method according to claim 11 , wherein in step g.), for the purpose of providing a population of ex vivo activated autologous plasma cells, the population resulting from step c.), after mixing with the population resulting from step e.), is further incubated with at least one of the cytokines of the group consisting of IL-6, IL-10, TNF-a, and sCD40L, preferably for 8-15 days, more preferably for 10 days.
  15. 15. Method according to one of claims 11-14, wherein step c.) includes providing an antigen preparation for the purpose of ex vivo activation of the autologous dendritic cells and autologous CD8+ cytotoxic T-lymphocytes in step d.) and for the purpose of ex vivo activation of autologous B lymphocytes in step e.).
  16. 16. Method according to claim 15, wherein, in case of an intended use of the medicament against a cancer disease of the human or animal patient, the antigen preparation is prepared by isolating antigens present on circulating tumor cells (CTCs) and/or cancer stem cells (CSCs) of the human or animal subject, wherein preferably, the circulating tumor cells and/or the cancer stem cells are isolated using magnetic cell separation, preferably using magnetic beads selective for recognition of at least one tumor antigen selected from a group consisting of CD44, CD133, EpCAM, PanCK.
  17. 17. Method according to one of claims 11-16, wherein in step a.), for the provision of a population of autologous macrophages which are polarized in a M1-phase, autologous monocytes are isolated and incubated in a medium free from GM-CSF and free from IL-4, wherein one day prior to step f.), IFN-y is added to the medium in order to trigger a polarization of the macrophages into M1 -stage.
  18. 18. Method according to one of claims 11-17, wherein for the purpose of providing the populations of steps a.)-e.), a sample of whole peripheral blood is collected from the human or animal patient, wherein preferably the populations resulting from steps a.)- e.) are derived from the same sample of whole peripheral blood.
  19. 9. Method of treating or preventing a disease in a human or animal subject, comprising the following steps: - administering a first dose comprising a first composition, said first composition comprising macrophages derived from autologous mononuclear cells, wherein the macrophages are preferably polarized in a M1 -phase, and further comprising autologous natural killer cells, preferably CD16+/CD56+ natural killer cells, to the human or animal subject; - administering a second dose comprising a second composition, said second composition comprising ex vivo activated autologous dendritic cells, ex vivo activated autologous CD8+ cytotoxic T-lymphocytes and ex vivo activated autologous plasma cells, to the human or animal subject; - administering a third dose comprising said second composition, to the human or animal subject; - wherein preferably the first dose, the second dose and the third dose are sequentially administered to the human or animal subject in intervals, preferably in intervals of at least one week, more preferably in intervals of 2 weeks.

Description

TITLE PHARMACEUTICAL PRODUCT FOR AUTOLOGOUS ADOPTIVE CELLULAR THERAPY TECHNICAL FIELD The present invention relates to a pharmaceutical product for autologous adoptive cellular therapy in the treatment of a cancer disease in a human or animal subject, comprising two compositions of various combinations of autologous immune cells to be administered sequentially in three doses to the human or animal subject, as well as a method for the manufacture of such a pharmaceutical product. PRIOR ART According to the WHO, cancer is the second leading cause of death worldwide, accounting annually for about 10 million deaths. Besides surgery, long-established and widely used conventional therapies, such as chemotherapies and radiation therapies, offer rapid results by killing cancer cells, but they have major drawbacks. In most cases, there is no specificity for cancerous cells and as a result, also healthy cells close to target cells are killed. Patients often face severe and long-lasting side effects, and furthermore, tumor cells can develop escape mechanisms and become resistant (Mansoori et al., 2017). It is crucial, especially for advanced-stage cancer patients, who have limited treatment options or have developed resistance to front-line therapies, to identify novel and effective therapies (Cheng et al., 2021). Immunotherapy triggers a patient's own immune system to fight cancer cells. It is increasingly gaining ground as a potential alternative treatment for cancer patients (Waldman et al., 2020), also due to the reduced side effects it results in. Targeted antibodies, checkpoint inhibitors, vaccines, cytokines, oncolytic viruses and adoptive cell transfer are different forms of established immunotherapies. They can be used alone or in combination with chemotherapies, radiation, surgery or targeted therapies to improve the efficacy of the treatment (Waldman et al., 2020). Adoptive cellular therapy is a type of immunotherapy which employs the use of ex vivo activated and expanded immune cells. Clinical trials have shown remarkable results of adoptive cellular therapies against tumors which are highly resistant to conventional therapies (Kirtane et al., 2021). Challenges limiting the efficiency of cellular therapies include immunosuppressive tumor microenvironment, insufficient number of tumor antigens, and poor cell trafficking. This is where personalized medicine steps in, by developing ways to determine, following a screening procedure, the potential of each patient to respond to a specific treatment. Each cancer type requires a specific treatment approach. Furthermore, as no two patients are the same, the treatment regimen shall be selected according to the genetic profile and disease history of the specific individual. In oncology, personalized treatment takes into account the inter- and intra-tumor variability of each individual patient (Hoeben et al., 2021). Efficiency of adoptive cellular immunotherapy depends on the immune status of each individual. Autologous cell therapies involve the isolation, activation and expansion of each patient's cells outside the body (ex vivo) and reintroduction of cells to the patient (Sarivalasis et al., 2021). Side effects generally are fewer than with other types of therapy. Innate immune cells are able to fight cancer cells in an antigen-independent manner, while T cells and B cells, being adoptive immune cells, require pre-activation with antigenic peptides. Activation of adoptive immune cells is performed with dendritic cells (DC), a type of antigen presenting cells (APC). Said immune cells can be generated from patient’s blood itself and then be infused back to the patient, enabling an autologous therapy. Macrophages (MP) and Natural Killer cells (NK) can recognize and kill cancer cells. Macrophages are phagocytic cells and natural killer cells which have the ability to eliminate abnormal cells, i.e. cancer cells in a manner independent of the major histocompatibility complexes class I or II (MHC-independent) (DeNardo et al., 2019; Multhoff et al., 2020; Kundu et al., 2021). DCs are the most efficient APCs and can activate both humoral and cellular immunity. DCs can be differentiated in vitro from their progenitors found in blood. The aim of in vitro manipulation of DCs for immunotherapeutical purposes, especially against cancer, is to obtain loading of both MHC class I and II molecules, for activation of both cellular and humoral immunity. Cytotoxic T-cells (CTLs) are activated against specific antigens by DCs and have the ability to target and kill cancer cells expressing the specific antigen (Hont et al., 2022; Parsonidis et al., 2019). Antibodies produced by plasma cells tag abnormal cells, especially cancer cells, and help their recognition by phagocytes or complement proteins, leading to the subsequent destruction of said targeted abnormal cells (Thomas et al. , 2020). Cancer cellular therapies up till now focused on single populations of immune cells, such as T-