US-12622935-B2 - Modified oncolytic adenoviruses

US12622935B2US 12622935 B2US12622935 B2US 12622935B2US-12622935-B2

Abstract

The invention concerns a modified replication competent, oncolytic adenovirus; a pharmaceutical composition comprising same; and a method of treating cancer using same.

Inventors

Tuuli Ranki
Sari PESONEN
Petri Priha
Erkko Ylösmäki
Vincenzo Cerullo
Beatriz Martins

Assignees

VALO THERAPEUTICS OY

Dates

Publication Date: 20260512
Application Date: 20190319
Priority Date: 20180321

Claims (11)

1 . A modified replicating adenovirus (Ad) of serotype 5 (Ad5) having lytic activity in target cancer cells comprising: a) a E1A gene deletion wherein the deletion is of nucleotides encoding amino acids 923-946 with respect to the wild-type Ad5 sequence; b) a 5/3 chimeric substitution of a knob of an adenoviral fiber protein wherein the knob of Ad5 is replaced by the knob of a serotype 3 Ad (Ad3); c) a 14.7k gene deletion wherein the deletion is of base pairs 30448-30834 with respect to the wild type Ad5 sequence and wherein the sequence GGA GGA GAT GAC TGA (SEQ ID NO: 1) is substituted for GGA GGA GAC GAC TGA (SEQ ID NO: 2); d) a gp19k gene deletion and a 7.1k gene deletion wherein the deletions are of base pairs 28541-29211 with respect to the wild type Ad5 sequence; and e) an insertion of a molecule encoding OX40L and a molecule encoding CD40L in the E3B region, replacing the 14.7K gene deletion, wherein the molecule encoding CD40L is inserted immediately downstream from the molecule encoding OX40L using a 2A processing site, and the 2A processing site is preceded by a cleavage site and a SGSG-linker (SEQ ID NO: 28) to ensure effective cleavage of OX40L and CD40L.
2 . The modified adenovirus according to claim 1 wherein said OX40L is human OX40L.
3 . The modified adenovirus according to claim 1 wherein said CD40L is human CD40L.
4 . The modified adenovirus according to claim 1 wherein said 2A processing site is a foot-and-mouth disease virus 2A processing site (F2A) or a porcine teschovirus-1 2A processing site.
5 . The modified adenovirus according to claim 1 wherein a molecule encoding the whole cDNA of each of OX40L and CD40L is inserted into said adenovirus.
6 . A pharmaceutical composition comprising at least one modified replication-competent and target cell lytic adenovirus according to claim 1 and a suitable carrier.
7 . The pharmaceutical composition according to claim 6 wherein said composition is formulated for intratumoral, intramuscular, intra-arterial, intravenous, intrapleural, intravesicular, intradermal, intracavitary or peritoneal injection, or an oral administration.
8 . A method of treating cancer in a patient comprising administering to a patient an effective amount of a composition comprising at least one modified replication-competent and target cell lytic adenovirus according to claim 1 .
9 . The method of treating cancer according to claim 8 wherein the at least one modified replication-competent and target cell lytic adenovirus is administered with a cell checkpoint modulator.
10 . The method of treating cancer according to claim 9 wherein the checkpoint modulator is an anti-PD1 molecule, an anti-PD-L1 molecule or an anti-CTLA-4 molecule.
11 . The method according to claim 8 wherein said cancer is selected form the list comprising or consisting of: nasopharyngeal cancer, synovial cancer, hepatocellular cancer, renal cancer, cancer of connective tissues, melanoma, lung cancer, bowel cancer, colon cancer, rectal cancer, colorectal cancer, brain cancer, throat cancer, oral cancer, liver cancer, bone cancer, pancreatic cancer, choriocarcinoma, gastrinoma, pheochromocytoma, prolactinoma, T-cell leukemia/lymphoma, neuroma, von Hippel-Lindau disease, Zollinger-Ellison syndrome, adrenal cancer, anal cancer, bile duct cancer, bladder cancer, ureter cancer, oligodendroglioma, neuroblastoma, meningioma, spinal cord tumor, osteochondroma, chondrosarcoma, Ewing's sarcoma, cancer of unknown primary site, carcinoid, carcinoid of gastrointestinal tract, fibrosarcoma, breast cancer, Paget's disease, cervical cancer, esophagus cancer, gall bladder cancer, head cancer, eye cancer, neck cancer, kidney cancer, Wilms' tumor, liver cancer, Kaposi's sarcoma, prostate cancer, testicular cancer, Hodgkin's disease, non-Hodgkin's lymphoma, skin cancer, mesothelioma, multiple myeloma, ovarian cancer, endocrine pancreatic cancer, glucagonoma, parathyroid cancer, penis cancer, pituitary cancer, soft tissue sarcoma, retinoblastoma, small intestine cancer, stomach cancer, thymus cancer, thyroid cancer, trophoblastic cancer, hydatidiform mole, uterine cancer, endometrial cancer, vagina cancer, vulva cancer, acoustic neuroma, mycosis fungoides, insulinoma, carcinoid syndrome, somatostatinoma, gum cancer, heart cancer, lip cancer, meninges cancer, mouth cancer, nerve cancer, palate cancer, parotid gland cancer, peritoneum cancer, pharynx cancer, pleural cancer, salivary gland cancer, tongue cancer and tonsil cancer.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This is the U.S. National Stage of International Application No. PCT/EP2019/056768, filed Mar. 19, 2019, which was published in English under PCT Article 21 (2), which in turn claims the benefit of Great Britain Application No. 1814867.6, filed Sep. 13, 2018, and Great Britain Application No. 1804473.5, filed Mar. 21, 2018. Incorporation of Electronic Sequence Listing The Sequence Listing is submitted as an ASCII text file, created on Sep. 17, 2020, 4.94 KB, which is incorporated by reference herein. FIELD OF THE INVENTION The invention concerns a modified replication competent, oncolytic adenovirus; a pharmaceutical composition comprising same; and a method of treating cancer using same. BACKGROUND OF THE INVENTION The perception of the role of oncolytic viruses in cancer treatment has changed dramatically during the last decade, as immunotherapy and the stimulation of the patient's own immune system to target and attack cancer has gained popularity. At the beginning of the century, oncolytic viruses were perceived as active agents in cancer treatment, acting solely through their inherent ability to lyse tumor cells via oncolysis. Recently, their use as cancer vaccines has gained interest, and their ability to release tumor antigens from cancer cells upon oncolysis for activating the immune system is recognised as an important characteristic in designing the ultimate immunotherapy against cancer. Adenoviruses are highly immunogenic viruses often used as vectors in various vaccine approaches against infectious diseases. Importantly, they have an exceptional ability to both prime and boost immune responses. Further, the presence of an oncolytic adenovirus within a tumor and the immunogenic cell death it causes is likely to shape the hostile tumor microenvironment towards a more susceptible state for a clinically relevant anti-tumor immunity to occur, by causing the expression of TH1-type immune modulators such as interferon gamma (IFNgamma). Immune cell infiltration to tumor is a frequent consequence of treatment with oncolytic viruses, and importantly, adenoviruses induce the infiltration by CD8+ T cells that are key effector cells in cancer immunity. Adenoviruses cause immunogenic cancer cell lysis whereupon tumor antigens, including unique patient specific neoantigens, previously hidden from the immune system or not presented in an immunogenic context are released into the immunogenic environment. This is the basis for a tumor-specific immune response caused by oncolytic adenoviruses. However, tumors have evolved several immunosuppressive mechanisms to counteract the immune cells of the body. Immune cells express cell surface molecules that regulate their activation and effector functions, specifically co-stimulatory and co-inhibitory molecules. The negative feedback molecules, i.e. checkpoint molecules, enable self-tolerance under normal physiological contexts, but are often utilized by the tumor cells to cause severe immune suppression. The best characterized checkpoint pathways are cytotoxic T-lymphocyte protein 4 (CTLA-4) and programmed cell death protein 1 pathway (PD-1/PD-L1). Due to these strong immunosuppressive mechanisms within the tumor, the virus-induced anti-tumor immune response can be weak unless strengthened by the use of immunostimulatory transgenes. The current approach tackles the problem of immune suppression at the tumor site, as the presence of the highly immunogenic oncolytic adenovirus encoding immunostimulatory human transgenes shapes the tumor microenvironment towards an “immune inflamed” phenotype that is more susceptible towards immunotherapy approaches. Importantly, the oncolytic virus of the current approach can be utilized in combination with checkpoint modulators such as anti-PD1, anti-PD-L1 or anti-CTLA-4 molecules to counteract immunosuppressive tumor environment and to cause a strong anti-immune response. STATEMENTS OF THE INVENTION According to a first aspect of the invention there is provided a modified, replicating adenovirus having lytic activity in target cancer cells comprising: a) a E1A gene deletion wherein the deletion is of nucleotides encoding amino acids 923-946;b) a 5/3 chimeric substitution of a knob of an adenoviral fiber protein wherein the knob of serotype 5 Ad is replaced by the knob of a serotype 3 Ad;c) a 14.7k gene deletion wherein the deletion is of base pairs 30448-30834 with respect to the wild type adenovirus and wherein the sequence GGA GGA GAT GAC TGA (SEQ ID NO: 1) is substituted for GGA GGA GAC GAC TGA (SEQ ID NO: 2); andd) a gp19k gene deletion and a 7.1k gene deletion wherein the deletions are of base pairs 28541-29211 with respect to the wild type adenovirus. In the above modified adenovirus amino acids 923-946 are deleted from the wild type (wt) Ad5 sequence. This deletion is a safety measure: as the viral E1A protein cannot bind to a retinoblastoma (Rb) molecule and release the transcription factor