Methods and products for treating or preventing lung cancer

Abstract

The present invention relates to methods for treating or preventing lung cancer in a subject, including a subject with Stage IV lung cancer. In particular, the methods include administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof to the subject. Also encompassed by the present inventio is a pharmaceutical composition when used for treating lung cancer in a subject, the composition including a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof. Combination products are also provided which include a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, and one or more of a chemotherapeutic agent, an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated, one or more immunotherapeutic agents, and one or more cancer cell-specific inhibitors. The combination product can be used to treat or prevent lung cancer, including Stage IV lung cancer.

Description

METHODS AND PRODUCTS FOR TREATING OR PREVENTING LUNG CANCERPRIORITY CLAIM

[0001] This application claims priority from Australian provisional patent application number 2023902817 filed on 1 September 2023, the contents of which are to be taken as incorporated herein by this reference.FIELD OF THE INVENTION

[0002] The present invention relates to methods and products for treating or preventing lung cancer in a subject.

[0003] The present invention further relates to methods for inhibiting formation and / or growth of tumours in a subject with lung cancer, improving the rate of survival / life expectancy of a subject with cancer, and inducing an anti-tumour immune response in a subject with lung cancer.

[0004] The present invention also relates to a method of selecting a subject with lung cancer suitable for treatment.BACKGROUND OF THE INVENTION

[0005] Lung cancer is a primary malignancy arising in the lungs and bronchi which can spread to pulmonary lymph nodes and within the lung(s) and metastasize elsewhere in the body to almost any region.

[0006] Lung cancer has been one of the most common cancers in the world for several decades. According to the World Cancer Research Fund International, it remains the second most common cancer worldwide, and indeed there were more than 2.2 million new cases of lung cancer diagnosed in 2020. In Australia alone, 13,078 new cases of lung cancer were diagnosed in 2018 with that number increasing in 2022. In the United States, an estimated 240,000 people will be diagnosed with lung cancer in 2023.

[0007] Lung cancer is also the most common cause of death from cancer, with 3 times as many men dying from lung cancer compared to prostate cancer, and 3 times as many women dying from lung cancer compared to breast cancer. According to the American Cancer Society, almost 130,000 people die annually in the US from lung cancer.

[0008] In human subjects, clinical staging of lung cancer is based on whether the tumor has spread to regional lymph nodes or distant sites. In Stage I lung cancer, the cancer cells are limited to the originating lung and so have yet to spread; however, in Stage II lung cancer nearby lymph nodes or the lining of the lungs will show signs of the cancer. In Stage III lung cancer, the cancer would have spread to the lymph nodes in the mediastinum or would have grown into the chest wall or the pericardium. Furthermore, the cancer may have spread to lymph nodes on the opposite side of the chest where the tumour first developed. The cancer may also have spread to the lower neck region. Subjects with Stage IV, or metastatic lung cancer, have cancer that has spread from its site of origin to distant lymph nodes and / or distant sites, such as bones, liver and brain. Further details regarding clinical staging of lung cancer is described in Nicholson AG at al., 2022, J. Thoracic Oncol., 17(3): 362-387, or can be found at the American Cancer Society (www.cancer.org / cancer / types / lung- cancer / detection-diagnosis-staging / staging-nsclc.html; and www.cancer.org / cancer / types / lung-cancer / detection-diagnosis-staging / staging-sclc.html).

[0009] The American Joint Committee on Cancer (AJCC) has designated staging by TNM classification to define lung cancer. For example, Stage IV lung cancer is defined by the following clinical stage grouping: any primary tumor (T), any metastasis to a regional lymph node (N), and a distant metastasis (M).

[0010] While 5-year survival rates for Stage I lung cancer are around 50% and for Stage II are around 35%, the survival rate for more advanced stages of lung cancer is poor. For example, AJCC Stage IV NSCLC has a dismal median survival time of around 4-months from diagnosis and a less than 10% 5-year survival rate. However, more advanced Stage IV lung cancer with widespread or rapidly appearing metastases has an even shorter survival time, with essentially a zero 5-year survival rate. Therefore, the aim of current treatments for metastatic NSCLC are to improve or maintain quality of life, and to prolong overall survival. Surgery for isolated metastases is an option for a subgroup of patients with Stage IV lung cancer, and occasionally chemotherapy can be effective in specific instances, but these options are usually not available for widespread metastatic disease and response rates to any form of standard therapy are almost non-existent.

[0011] Despite many studies relating to the development of therapeutic interventions for the treatment of lung cancer, there has been little progress in this field. Indeed, apart from surgical intervention and the use of chemotherapeutic agents, which have a low percentage impact, there is no effective treatment for Stage IV lung cancer.

[0012] In light of above, there is a real and present need for new treatments for lung cancer, including Stage IV lung cancer.

[0013] The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any, or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.SUMMARY OF THE INVENTION

[0014] The present invention is predicated in part on the identification of a novel treatment method for lung cancer. In this regard, in a first aspect the present invention provides a method of treating or preventing lung cancer in a subject, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. In some embodiments, the subject is a human.

[0015] In some embodiments, the melanoma cell lysate includes fragmented melanoma cell membranes. In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate. In some embodiments, the melanoma cell lysate is an allogeneic cell lysate. In some embodiments, the melanoma cell lysate includes a lysate from MM200 cells.

[0016] In some embodiments, the lung cancer is Stage IV lung cancer. In some embodiments, the Stage IV lung cancer is advanced Stage IV lung cancer.

[0017] In some embodiments, the administration of the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, to the subject includes multiple administrations of the melanoma cell lysate to the subject during a treatment period. In some embodiments, the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for one or more cycles. In some embodiments, the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for at least about one month. In some embodiments, the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for at least about three months.

[0018] In some embodiments, the subject undergoes radiation therapy and / or chemotherapy and / or surgery before, during or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0019] In some embodiments, the method includes the further step of administering to the subject one or more of: one or more chemotherapeutic agents; one or more immunotherapeutic agents; and one or more cancer cell-specific inhibitors, before, during or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. In some embodiments, the one or more immunotherapeutic agents comprise one or more immune checkpoint inhibitors. In some embodiments, the method includes the further step of one or more administrations of an effective amount of an agent to the subject that reduces the level and / or activity of regulatory T cells in the subject.

[0020] In some embodiments, the method includes the step of selecting a subject suitable for treatment. In some embodiments, the subject is selected on the basis that the subject shows a T cell response to a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. In some embodiments, the subject is selected on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administration to the subject of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0021] In some embodiments, formation and / or growth of a secondary tumour in the subject is inhibited. In some embodiments, growth of a primary lung cancer in the subject is inhibited. In some embodiments, rate of survival of the subject is improved. In some embodiments, life expectancy of the subject is improved. In some embodiments, an anti-tumour immune response in the subject is induced. In some embodiments, one or more of an increase in the number of intratumoral dendritic cells is produced in the subject, dendritic cell activation is improved in the subject, and the number and / or activity of regulatory T cells is decreased in the subject.

[0022] In a second aspect, the present invention provides a method of inhibiting formation and / or growth of a secondary tumour in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0023] In a third aspect, the present invention provides a method of inhibiting growth of a primary lung cancer in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0024] In a fourth aspect, the present invention provides a method of improving the rate of survival of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0025] In a fifth aspect, the present invention provides a method of increasing life expectancy of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0026] In a sixth aspect, the present invention provides a method of inducing an anti-tumour immune response in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0027] In some embodiments of the second to sixth aspects of the invention, the subject is a human.

[0028] In some embodiments of the second to sixth aspects of the invention, the melanoma cell lysate includes fragmented melanoma cell membranes. In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate.

[0029] In some embodiments of the second to sixth aspects of the invention, the lung cancer is Stage IV lung cancer.

[0030] In a seventh aspect, the present invention provides use of a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, in the preparation of a medicament for treating lung cancer in a subject.

[0031] In some embodiments of the seventh aspect of the invention, the subject is a human.

[0032] In some embodiments of the seventh aspect of the invention, the melanoma cell lysate includes fragmented melanoma cell membranes. In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate.

[0033] In some embodiments of the seventh aspect of the invention, the lung cancer is Stage IV lung cancer.

[0034] In an eighth aspect, the present invention provides a pharmaceutical composition when used for treating lung cancer in a subject, the composition including a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof.

[0035] In a ninth aspect, the present invention provides a combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and00 a chemotherapeutic agent; and / or (ill) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

[0036] In a tenth aspect, the present invention provides a combination product when used for treating or preventing lung cancer in a subject, the combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and00 a chemotherapeutic agent; and / or (iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

[0037] In some embodiments of the ninth or tenth aspects of the invention, the components are provided in a form for separate administration to the subject.

[0038] In some embodiments of the eighth to tenth aspects of the invention, the melanoma cell lysate includes fragmented melanoma cell membranes. In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate. In some embodiments, the melanoma cell lysate is an allogeneic cell lysate. In some embodiments, the melanoma cell lysate includes a lysate from MM200 cells. In some embodiments, the one or more immunotherapeutic agents comprise one or more immune checkpoint inhibitors.

[0039] In some embodiments of the eighth to tenth aspects of the invention, the lung cancer is Stage IV lung cancer.

[0040] In some embodiments of the eighth to tenth aspects of the invention, the composition or combination product inhibits growth of a primary lung cancer in the subject. In some embodiments, the composition or combination product inhibits formation and / or growth of a secondary tumour in the subject. In some embodiments, the composition or combination product improves rate of survival of the subject. In some embodiments, the composition or combination product improves life expectancy of the subject. In some embodiments, the composition or combination product induces an anti-tumour immune response in the subject.

[0041] In some embodiments of the ninth or tenth aspects of the invention, the combination product includes instructions for selecting a subject with lung cancer suitable for treatment. In some embodiments, the product includes instructions for selecting a subject with lung cancer suitable for treatment on the basis that the subject shows a T cell response to the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. In some embodiments, the product includes instructions for selecting a subject with lung cancer suitable for treatment on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administration to the subject of the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof.

[0042] In an eleventh aspect, the present invention provides a method of treating lung cancer in a human subject, the method including the step of administering to the subject a human vaccinina melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0043] In a twelfth aspect, the present invention provides a method of selecting a subject with lung cancer suitable for treatment with a melanoma cell lysate and / or animmunotherapeutic extract, component or antigen thereof, the method including the step of identifying a subject that shows a T cell response upon vaccination of the subject with the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0044] In a thirteenth aspect, the present invention provides a method of selecting a subject with lung cancer suitable for treatment with a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof, the method including the step of identifying a subject that has a decreased level and / or activity of regulatory T cells upon vaccination of the subject with a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0045] In some embodiments of the eleventh to thirteenth aspects of the invention, the lung cancer is Stage IV lung cancer.DETAILED DESCRIPTION OF THE INVENTION

[0046] The present invention arises from the effective treatment of subjects with lung cancer using a vaccine produced from melanoma cells (a melanoma cell lysate).

[0047] Accordingly, certain disclosed embodiments provide methods, compositions, and combination products, that have one or more advantages. For example, some of the advantages of some embodiments disclosed herein include one or more of the following: new methods, compositions, and combination products to treat lung cancer; new methods, compositions, and combination products to prevent lung cancer; new methods, compositions, and combination products to inhibit formation and / or growth of a secondary tumour; new methods, compositions, and combination products to inhibit growth of a primary lung cancer; new methods, compositions, and combination products to increase the rate of survival of a subject with lung cancer; new methods, compositions, and combination products to increase life expectancy of a subject with lung cancer; to provide one or more advantages, or to provide a commercial alternative. Other advantages of some embodiments of the present disclosure are provided herein.

[0048] In a first aspect the present invention provides a method of treating or preventing lung cancer in a subject, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0049] In a particular embodiment, a Vaccinia Melanoma Cell Lysate (VMCL) vaccine has been used for the treatment. In this embodiment, the cell lysate was prepared from a melanoma cell line. It is shown herein that the VMCL vaccine is effective in inducing complete regression of tumours in a patient with advanced "incurable" Stage IV lung cancer. Use of a melanoma cell lysate also provides a non-toxic avenue for treatment of patients with Stage IV lung cancer.

[0050] The term "subject" as used throughout the specification is to be understood to mean any human or animal subject. In this regard, the present invention includes within its scope veterinary applications for the treatment of melanoma. For example, the animal subject may be a mammal, a primate, a livestock animal (eg. A horse, a cow, a sheep, a pig, or a goat), a companion animal (eg. A dog, a cat), a laboratory test animal (eg. A mouse, a rat, a guinea pig, a bird), an animal of veterinary significance, or an animal of economic significance.

[0051] It will also be appreciated that the term "subject" includes within its scope a subject originally diagnosed as suffering from a lung cancer and who has been subsequently treated for the cancer using current treatments (such as chemotherapy, radiation therapy, immune checkpoint inhibitor therapy, cancer cell-specific inhibitor therapy, surgery, and the like). Therefore, the present invention extends to the treatment of those original subjects using a method as described herein. For example, the subject may be a human subject suffering from a lung cancer treated for the cancer and then subsequently treated using a method as described herein.

[0052] The terms "treat" and "treating", and variants thereof, as used throughout the present specification are to be understood to mean therapeutic intervention with a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) of the present invention. The terms "prevent", "preventing" and "prevention" as used herein are to be understood to include within their scope obtaining a desired pharmacologic and / or physiologic effect in terms of arresting or suppressing the appearance of one or more symptoms in the subject.

[0053] For example, the aforementioned terms include within their scope a therapeutic or preventative intervention which has one or more of the following outcomes in the subject: (i) inhibiting or preventing the growth of a primary lung cancer in the subject, including reducing the growth of the primary tumour after resection; (ii) inhibiting or preventing the formation and / or growth of one or more secondary tumours in the subject, including tumours in thelymph nodes; (ill) inhibiting the recurrence of the cancer in the subject following treatment; (iv) increasing the life expectancy of the subject as compared to the untreated state; (v) improving the rate of survival of the subject; (vi) inducing an anti-tumour immune response in the subject; and (vii) improving the quality of life the subject as compared to the untreated state. Accordingly, certain disclosed embodiments of the present invention provide methods and compositions that have one or more advantages, and / or provide a commercial alternative to existing methods and compositions. Other advantages of some embodiments of the present disclosure are provided herein.

[0054] The term "inhibit" and the like as used throughout the specification is to be understood to mean a reduction in the progress of a process, including any one or more of the start, continuation or termination of a process.

[0056] The present invention is directed to a method of treating lung cancer using an immunotherapeutic approach, and in particular, treating the cancer by administering a vaccine derived from melanoma cells. In humans, assessment of the clinical staging of lung cancer may generally be made by a suitably qualified practitioner using any tests that are generally known and accepted in the art. In some embodiments, the cancer staging can comprise that developed by the American Joint Committee on Cancer (AJCC) which has designated staging in humans by “TNM" classification to define lung cancer. Generally, the TNM system provides results from various tests and scans in order to determine the size and location of the primary tumor (Tumor, I); whether the cancer has spread to the lymph nodes, and if it has, the location and number of the affected lymph nodes (Node, N); and whether the cancer has spread to other parts of the body, and if it has, the extent and location of the remote cancer (Metastasis, M). While each type of cancer may have its own specific system, the TNM staging system generally uses scaled scoring for each letter.

[0056] For Tumor, “T is associated with a number (e.g., 0 to 4) to describe the general tumor size, location, and whether it intrudes into nearby tissues. Larger or more intrusive tumors are given a higher number and, depending on the cancer, a lowercase letter, such as "a," “b,” or “m" (for multiple), may be added to provide more detail. Similarly, for Node, “N" is associated with a number (e.g., 0 to 3) to describe whether cancer has been found in the lymph nodes, and can also indicate the number of lymph nodes containing cancer. Larger numbers are assigned when more lymph nodes are involved with cancer. For Metastasis, "M" indicates whether or not the cancer has spread to other parts of the body and is labeled MO for no spread, or M1 if it has spread.

[0057] The T, N, and M results are combined to determine the stage of cancer, typically one of four stages: stages I (one) to IV (four). Some cancers also have a stage 0 (zero). Stage 0 describes cancer in situ, remaining local to the original tissue without any spread to nearby tissues. Staging may include optional analysis of prognostic factors to provide chances of recovery and a recommended therapy. Prognostic factors may include grading the cancer based on appearance of the cancer cells; analysis of tumor marker expression; and analysis of tumor genetics. A cancer may be restaged using the same initial system in order to determine efficacy of a treatment or obtain more information about a recurrent cancer.

[0058] In some embodiments of the present invention, the lung cancer is non-small cell lung cancer (NSCLC) or is small cell lung cancer (SCLC). In some embodiments, the lung cancer is Stage III lung cancer. In some embodiments of the present invention, the lung cancer is Stage IV lung cancer. Stage IV lung cancer is associated with the spread to more than one area in the other lung, the fluid surrounding the lung or the heart, or distant metastasis in the body. Lung cancer is more likely to spread to the brain, bones, liver, and adrenal glands. Stage IV lung cancer includes substages IVA (spread within the chest) and IVB (spread outside of the chest). Surgery is rarely successful for most stage IV lung cancers and may be impossible to remove if it has spread to the lymph nodes above the collarbone, or to vital structures within the chest (e.g., heart, large blood vessels, or the main pulmonary structures).

[0059] In some embodiments of the present invention, the Stage IV lung cancer is advanced Stage IV lung cancer. Advanced Stage IV lung cancer is generally non-resectable or where surgery is not able to be performed, whereas Stage IV lung cancer may be resectable.

[0060] The melanoma cell lysate in the various forms of the present invention is a lysate that is able to induce an anti-tumour response in the subject, so as to provide an immunotherapeutic effect in the subject. In this regard, the term "lysate" is to be understood to mean the cellular debris and / or fluid produced by lysis of a cell, or an extract, a component including a semi-purified or purified component, or antigen derived from the lysate. The immunotherapeutic approach of the present invention includes administration of one or more of a lysate, an extract of the lysate, a semi-purified or purified component derived from the lysate, and one or more immunotherapeutic antigens derived from the lysate.

[0061] Lysis of a melanoma cell may be achieved by a suitable method, for example, by viral lysis. Methods for producing aann immunotherapeutic extract of the lysate, animmunotherapeutic component of the lysate (semi-pruified or purified), or one or more antigens present in the lysate with an immunotherapeutic effect are known in the art.

[0062] In some embodiments of the present invention, the melanoma cell lysate includes fragmented melanoma cell membranes.

[0063] It is appreciated that the lysate in the various forms of the present invention includes either or both of antigens that are, or were derived from, membrane-associated and non- membrane associated antigens. In this regard, the term "membrane-associated" will be understood to mean an antigen that normally is part of a membrane from a melanoma cell, an antigen that is normally bound to a membrane from a melanoma cell, or an antigen that co-purifies with a cell membrane so as to be associated with a cell membrane in the lysate of the present invention.

[0064] In some embodiments of the present invention, the lysate includes membrane- associated antigens.

[0066] In some embodiments, the melanoma cell lysate is an allogeneic cell lysate. For example, in the case of treating a human subject, the melanoma cell lysate may be an allogeneic human melanoma cell lysate.

[0066] In the case where the lysate includes fragmented cell membranes, the presence of fragmented cell membranes in the melanoma cell lysate may be determined by a suitable method known in the art. For example, the presence of fragmented cell membranes in the lysate may be confirmed by microscopic analysis of the lysate.

[0067] In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In this regard, the virus used to produce the lysate may be a naturally occurring strain (or a derivative thereof), or may be a recombinant vims. In the case of a recombinant vims, the recombinant vims may also encode one or more gene products. For example, the recombinant vims may encode an immunostimulating molecule such as a cytokine, a hematopoietic growth factor or a melanoma immunogen.

[0068] Methods for producing cell lysates with viruses are known in the art. Generally, to prepare a viral melanoma cell lysate, melanoma cells are infected with a suitable virusresulting in lysis of a proportion of the cells. After a suitable period of time, the lysed cells are collected and processed to produce a lysate suitable for administration to a subject.

[0069] For example, the cells may be collected, homogenised and subject to centrifugation to produce a supernatant. The pellet resulting after centrifugation may be also further processed, such as by being subjected to one or more freeze-thaw cycles, and combined with the earlier supernatant. The resulting supernatant (either from the centrifugation or when combined with the freeze-thawed cells) may then be subjected to a further high-speed centrifugation to sediment the material. The pellet may then be resuspended in saline and the protein and viral content determined. The lysate is then in a form suitable for administration to a subject. The lysate may be further processed to produce an extract of the lysate, a component of the lysate, or purify one or more antigens present in the lysate, each with immunotherapeutic effect.

[0070] In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate. Methods for producing cell lysates with vaccinia virus are known in the art.

[0071] Accordingly, in a further aspect the present invention provides a method of treating lung cancer in a human subject, the method including the step of administering to the subject a human vaccinia melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. In some embodiments, the lung cancer is Stage VI lung cancer.

[0072] In the case where a recombinant virus is used to prepare the cell lysate, the virus may also encode an antigen such as a melanoma immunogen (e.g. MAGE-1 , MAGE-3, SAGE, GAGE, FRAME and NY-ESO-1); melanocyte differentiation antigens (e.g. tyrosinase, Melan- A / MART-1, gp100, TRP-1 and TRP-2); mutated or aberrantly expressed antigens (e.g. MUM- 1 , CDK4, beta-catenin, gp100 and N-acetylglucosaminyltransferase); and other antigens like B7-1 , TA-90, lysosome-associated membrane protein (LAMP), melanocyte-stimulating hormone receptor (MCIR), and p90 calnexin.

[0073] Methods for producing recombinant viruses encoding a desired gene product are known in the art. Methods for cloning nucleic acids into viral vectors are known in the art, for example as described in Green MR and Sambrook J, Molecular Cloning: A Laboratory Manual (4thedition), Cold Spring Harbor Laboratory Press, 2012.

[0074] As discussed above, the melanoma cell lysate may be an allogeneic melanoma cell lysate. However, it will be appreciated that the melanoma cell lysate in the various forms of the present invention may also be a lysate produced (or derived) from melanoma cells or cell lines (primary or secondary) from the subject being treated (i.e. an autologous cell lysate), including melanoma cells derived from the primary tumour or a metastasis resected from the patient.

[0075] Methods for producing an autologous cell lysate are known in the art. For example, a suitable method for isolating a malignant cell suspension for use as an autologous antigen source is by excision of malignant tissue and mechanically disaggregated using a scalpel in RPMI 1640 medium in a Petri dish, a flask, or a vessel. The cell suspension may be rinsed with RPMI 1640 and transferred to a sterile tissue culture plate, RPMI 1640 containing 5% FCS and antibiotics added and cells cultured in a 37°C, 5% CO2humidified incubator.

[0076] Primary or secondary cell lines may also be produced from these cells by a method known in the art. Procedures for establishing melanoma cell lines are known in the art.

[0077] In the case of treatment of a human subject with the melanoma cell lysate of the present invention, the melanoma cell lysate may be for example an allogeneic cell line referred to as MM200, as described in Pope et al., 1979, Pathology, 11: 191-195. This cell line has the HLA type A1 ,3; B7.35; DR2.4. Chromosomal analysis of this cell line showed a nodal number of 76, and a number of marker chromosomes were also revealed by karyotypic analysis (Muir and Gunz, 1979, Pathology, 11: 597-606). Antigens known to be expressed in MM200 cells are Tyrosinase, gp100; MART-1; MAGE-A3; MAGE-A10; BAGE; GAGE; XAGE.

[0078] In some embodiments, the melanoma cell lysate of the present invention includes a lysate (and / or an immunotherapeutic extract, component or antigen thereof) from MM200 cells.

[0079] Other human melanoma cell lines are described in Satyamoorthy et al., 1997, Melanoma Res., 7 Suppl 2: S35-42.

[0080] Human melanoma cells are also available from from the American Tissue Culture Collection. Examples of a non-limiting list of such cells are shown in Table 1.TABLE 1ATCC Melanoma Cell lines

[0081] In one form, the melanoma cells used to produce the lysate in the various forms of the present invention provide at least one NLA class I antigen.

[0082] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject in the absence of an adjuvant. As would be understood in the art, an adjuvant is any agent that acts to boost or augment an immune response with the aim of making the melanoma cell lysate more effective, or more active, in its intended action.

[0083] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject in the absence of a carrier, helper protein, or hapten.

[0084] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), when administered to the subject, may include one or more adjuvants, carriers, helper proteins, haptens, and / or excipients, as described in more detail below. An example of an adjuvant is montanide.

[0085] The administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to the subject in the various forms of the present invention may be performed by a suitable method known in the art and may be conducted over a suitable period of time.

[0086] In some embodiments, administration is performed using multiple injections administered over a time course which is selected to maximize an immune response in the subject. Accordingly, administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to the subject may include multiple administrations to the subject during a treatment period prior to administration of the immune checkpoint inhibitor to the subject.

[0087] In some embodiments of the methods encompassed herein, administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to the subject includes a treatment period of administration about every one to four weeks for one or more cycles. In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject about every one to four weeks for at least about one to six months. In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject about every two weeks for at least about one month. In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject about every two weeks for at least about three months. In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject about every two weeks for at least about three to six months. In some embodiments, administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to the subject includes a treatment period of administration about every two weeks for one or more cycles, followed by a period of administration about once a month for one or more cycles. In some embodiments, administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to the subject includes a treatment period of administration about every two weeks for one or more cycles, followed by a period of administration about once a month for one or more cycles, followed by a period of administration about once every three months for one or more cycles.

[0088] In some embodiments, the subject receives bi-weekly administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) (e.g. by injection) for about six weeks, then about every month for about six months, then about every three months for about six months. However, any suitable dosing regimen of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) can be used as would be determined by a skilled physician.

[0089] It will be appreciated that administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is not to be restricted to injection, and indeed the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), may be administered by any conventional route including parenteral and oral routes, as explained in more detail below. Examples of parenteral routes include subcutaneous, intradermal, transcutaneous, intra-tumoural, intravascular, intravenous, intramuscular, intraorbital, intracapsular, intrathecal, intraspinal, intracranial, intraventricular,intracistemal, intranasal and intraperitoneal. In the case of injection, suitable sites of the injection in a human subject are on anterior thighs, anterior upper arms, or the anterior thorax

[0090] It will also be appreciated that the administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) also includes within its scope ex vivo treatment of the subject’s cells, or a suitably matched donor’s cells, and re- introduction into the subject. For example, antigen presenting cells such as dendritic cells may be isolated from the subject and treated or contacted with the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). Methods for isolating antigen presenting cells, such as dendritic cells, are known in the art.

[0091] The antigen presenting cells discussed above include antigen presenting cells isolated from a subject, or an antigen presenting cell formed in vitro from a precursor cell. The antigen presenting cells may be present in a mix of one or more types of cell, or alternatively may be substantially purified from other types of cells.

[0092] For example, dendritic cells may be isolated directly from a subject, as a mixture of cells enriched for dendritic cells prepared by leukapheresis or separation of peripheral blood from a subject. Alternatively, the isolated dendritic cells may be prepared by obtaining dendritic cell precursor cells and treating them in vitro to form immature dendritic cells, for example as described in Sallustro et al., 1994, J. Exp. Med., 179: 1109-18.

[0093] The introduction of treated antigen presenting cells into a subject may be by a suitable method known in the art, including the introduction intravenously into the subject. For example, treated dendritic cells may be introduced into a human subject as described in Lau et al., 2001, J. Immunol., 24(1): 66-78. Typically, 1x10® to 1x10® dendritic cells will be introduced into a human subject. The time period over which introduction of the treated cells occurs will depend on a number of factors, including the extent of the immune response required, the age and body weight of the subject, and the administration of other active agents to the subject.

[0094] The dosage levels of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) administered to the subject in the various forms of the present invention are not particularly limited and depend on the mode of administration, thecharacteristics of the subject, and the nature of any carrier or adjuvant that may be included in a formulation.

[0095] It will be appreciated that the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject in an effective amount. The term "effective amount" as used herein is the quantity of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) which, when administered to a subject, improves the prognosis and / or health state of the subject. The amount of melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) to be administered to a subject will depend on particular characteristics as intimated above, including (but not limited to) one or more of: the extent of the immune response required; the mode of administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof); and the characteristics of the subject, such as general health, other diseases, age, sex, genotype, and body weight. A person skilled in the art will be able to determine appropriate dosages depending on these and other factors.

[0096] In the case of a vaccinia melanoma cell lysate, a suitable dose for administration is in the range of 0.1-1.0 ml lysate. A suitable dose may therefore include, but is not limited to, 0.1-0.2 ml lysate, 0.1-0.3 ml lysate, 0.1-0.4 ml lysate, 0.1-0.5 ml lysate, 0.1-0.6 ml lysate, 0.1- 0.7 ml lysate, 0.1-0.8 ml lysate, 0.1-0.9 ml lysate, 0.1-1.0 ml lysate, 0.2-0.3 ml lysate, 0.2-0.4 ml lysate, 0.2-0.5 ml lysate, 0.2-0.6 ml lysate, 0.2-0.7 ml lysate, 0.2-0.8 ml lysate, 0.2-0.9 ml lysate, 0.2-1.0 ml lysate, 0.3-0.4 ml lysate, 0.3-0.5 ml lysate, 0.3-0.6 ml lysate, 0.3-0.7 ml lysate, 0.3-0.8 ml lysate, 0.3-0.9 ml lysate, 0.3-1.0 ml lysate, 0.4-0.5 ml lysate, 0.4-0.6 ml lysate, 0.4-0.7 ml lysate, 0.4-0.8 ml lysate, 0.4-0.9 ml lysate, 0.4-1.0 ml lysate, 0.5-0.6 ml lysate, 0.5-0.7 ml lysate, 0.5-0.8 ml lysate, 0.5-0.9 ml lysate, 0.5-1.0 ml lysate, 0.6-0.7 ml lysate, 0.6-0.8 ml lysate, 0.6-0.9 ml lysate, 0.6-1.0 ml lysate, 0.7-0.8 ml lysate, 0.7-0.9 ml lysate, 0.7-1.0 ml lysate, 0.8-0.9 ml lysate, 0.8-1.0 ml lysate, or 0.9-1.0 ml lysate. In some embodiments, 0.3 ml of the lysate is administered to the subject per dose.

[0097] The manner in which the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) is administered to the subject is not limited to a particular mode. In this regard, one route of administration may be systemic administration and therefore the lysate may be in the form of an injectable solution, or may be in another dosage form useful for systemic administration of agents. Another route of administration may include topical administration and therefore the lysate, or a composition comprising thelysate, may be in the form of a liquid, gel, suspension, paste, lotion, cream, solid, semi-solid, powder, and the like. Other forms of administration may include delivery by way of a scaffold, such as a biomaterial scaffold including a scaffold produced from collagen, hydroxyapatite, β- tricalcium phosphate or a combination thereof. Other routes of administration are contemplated.

[0098] The melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) may be administered in the form of a suitable pharmaceutical composition, for example in a mixture with other therapeutic substances and / or other substances that enhance, stabilise or maintain the activity of the active component of the pharmaceutical composition. In some embodiments, an administration vehicle (e.g., injectable solution, liquid, gel, paste, powder, cream, pill, tablet, capsule, aerosol, etc) would contain the active component or a pharmaceutical composition comprising the active component and / or additional substance(s). In this regard, the pharmaceutical composition may also include the use of one or more pharmaceutically acceptable carriers or additives, including pharmaceutically acceptable salts, amino adds, polypeptides, polymers, solvents, buffers, exdpients and bulking agents, taking into consideration the particular physical and chemical characteristics of the active component to be administered.

[0099] In some embodiments, the carrier may be chosen based on various considerations including the route of administration, and the time course of delivery of the pharmaceutical composition. The term "pharmaceutically acceptable carrier" refers to a substantially inert solid, semi-solid or liquid filler, diluent, excipient, encapsulating material or formulation auxiliary of any type. An example of a pharmaceutically acceptable carrier is physiological saline. Other physiologically acceptable carriers and their formulations are known in the art. Some examples of materials which can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose and sucrose; starches such as com starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; com oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; detergents such as TWEEN 80; buffering agents such as magnesium hydroxide and aluminium hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions, as well as other non- toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colouring agents, releasing agents, coating agents, sweetening, flavouring and perfumingagents, preservatives and antioxidants can also be present.

[0100] The preparation of such pharmaceutical compositions is known in the art, for example as described in Remington's Pharmaceutical Sciences, 18th ed., 1990, Mack Publishing Co., Easton, Pa. and U.S. Pharmacopeia: National Formulary, 1984, Mack Publishing Company, Easton, Pa, which are incorporated herein by reference in their entirety.

[0101] As intimated above, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may be formulated for parenteral administration. The term ''parenteral'' as used herein includes, but is not limited to, subcutaneous, intradermal, transcutaneous, intravascular, intravenous, intramuscular, intraorbital, intracapsular, intrathecal, intraspinal, intracranial, intraventricular, intracistemal, intranasal and intraperitoneal injection or infusion techniques.

[0102] When administered parenterally, tthhee melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, will normally be in a unit dosage, sterile injectable, form (solution, suspension or emulsion) which is preferably isotonic with the blood of the recipient with a pharmaceutically acceptable carrier. Examples of such sterile injectable forms are sterile injectable aqueous or oleaginous suspensions. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable forms may also be sterile injectable solutions or suspensions in non-toxic parenterally-acceptable diluents or solvents, for example, as solutions in 1 ,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, saline, Ringer's solution, dextrose solution, isotonic sodium chloride solution, and Hanks' solution. In addition, sterile, fixed oils are conventionally employed as solvents or suspending mediums. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides, com, cottonseed, peanut, and sesame oil. Fatty acids such as ethyl oleate, isopropyl myristate, and oleic acid and its glyceride derivatives, including olive oil and castor oil, especially in their polyoxyethylated versions, are useful in the preparation of injectables. These oil solutions or suspensions may also contain long- chain alcohol diluents or dispersants.

[0103] The carrier may contain minor amounts of additives, such as substances that enhance solubility, isotonicity, and chemical stability, for example anti-oxidants, buffers and preservatives.

[0104] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may be formulated for topical administration, e.g. transdermal administration. Transdermal administrations are understood to include all administrations across the surface of the body and the inner linings of bodily passages including epithelial and mucosal tissues. Such administrations may be carried out using a liquid, gel, paste, lotion, cream, ointment, powder, foam, patch, suspension, solution, or other suitable formulation.

[0105] A cream is a formulation that contains water and oil and is stabilized with an emulsifier. Lipophilic creams are called water-in-oil emulsions, and hydrophilic creams oil-in- water emulsions. The cream base for water-in-oil emulsions are normally absorption bases such as vaseline, ceresin or lanolin. The bases for oil-in-water emulsions are mono-, di-, and tri-glycerides of fatty acids or fatty alcohols with soaps, alkyl sulphates or alkyl polyglycol ethers as emulsifiers.

[0106] A lotion is an opaque, thin, non-greasy emulsion liquid dosage form for external application to the skin, which generally contains a water-based vehicle with greater than 50% of volatiles and sufficiently low viscosity that it may be delivered by pouring. Lotions are usually hydrophilic and contain greater than 50% of volatiles as measured by LOD (loss on drying). A lotion tends to evaporate rapidly with a cooling sensation when rubbed onto the skin.

[0107] A paste is an opaque or translucent, viscous, greasy emulsion or suspension semisolid dosage form for external application to the skin, which generally contains greater than 50% of hydrocarbon-based or a polyethylene glycol-based vehicle and less than 20% of volatiles. A paste contains a large proportion (20-50%) of dispersed solids in a fatty or aqueous vehicle.

[0108] An ointment is an opaque or translucent, viscous, greasy emulsion or suspension semisolid dosage form for external application to the skin, which generally contains greater than 50% of hydrocarbon-based or a polyethylene glycol-based vehicle and less than 20% of volatiles. An ointment is usually lipophilic and contains >50% of hydrocarbons or polyethylene glycols as the vehicle and <20% of volatiles as measured by LOD. An ointment tends not to evaporate or be absorbed when rubbed onto the skin.

[0109] A gel is usually a translucent, non-greasy emulsion or suspension semisolid dosage form for external application to the skin, which contains a gelling agent in quantities sufficient to impart a three-dimensional, cross-linked matrix A gel is usually hydrophilic and contains sufficient quantities of a gelling agent such as starch, cellulose derivatives, carbomers, magnesium-aluminum silicates, xanthan gum, colloidal silica, aluminium or zinc soaps.

[0110] The melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, when in a form for topical administration, may further include drying agents, anti-fbaming agents, buffers, neutralizing agents, agents to adjust pH, colouring agents and decolouring agents, emollients, emulsifying agents, emulsion stabilizers and viscosity builders, humectants, odorants, preservatives, antioxidants, and chemical stabilizers, solvents, and thickening, stiffening, and suspending agents, and a balance of water or solvent.

[0111] Transdermal administration may also be accomplished through the use of a transdermal patch containing the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), and a carrier that is inert to the active agents, is non- toxic to the skin, and allows delivery of the active agents for systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active agents may also be suitable. A variety of occlusive devices may be used to release the active agents into the blood stream such as a semi-permeable membrane covering a reservoir containing the active agents with or without a carrier, or a matrix containing the active agents. Transdermal formulations are known in art and may be formulated by a skilled person.

[0112] The melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may also be formulated using controlled release technology. For example, they may be administered as a sustained-release pharmaceutical. To further increase the sustained release effect, they may be formulated with additional components such as vegetable oil (for example soybean oil, sesame oil, camellia oil, castor oil, peanut oil, rape seed oil); middle fatty acid triglycerides; fatty acid esters such as ethyl oleate; glycerol monooleate; polysiloxane derivatives; alternatively, water-soluble high molecular weight compounds such as hyaluronic acid orsalts thereof (weight average molecular weight: ccaa.. 80,000 ttoo 2,000,000), carboxymethylcellulose sodium (weight average molecular weight: ca. 20,000 to 400,000), hydroxypropylcellulose (viscosity in 2% aqueous solution: 3 to 4,000 cps), atherocollagen (weight average molecular weight: ca. 300,000), polyethylene glycol (weight average molecular weight: ca. 400 to 20,000), polyethylene oxide (weight average molecular weight: ca. 100,000 to 9,000,000), hydroxypropylmethylcellulose (viscosity in 1% aqueous solution: 4 to 100,000 cSt), methylcellulose (viscosity in 2% aqueous solution: 15 to 8,000 cSt), polyvinyl alcohol (viscosity: 2 to 100 cSt), polyvinylpyrrolidone (weight average molecular weight: 25,000 to 1,200,000).

[0113] Alternatively, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may be incorporated into a hydrophobic polymer matrix, scaffold or support (such as a biodegradable matrix or support), including for controlled release of the active agent over a period of days. Methods for delivering agents via scaffolds are known in the art. For example, a biomaterial scaffold including a scaffold produced from collagen, hydroxyapatite, β- tricalcium phosphate or a combination thereof may be used to deliver the active agent. Alternatively, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), or a pharmaceutical composition comprising said agent may be incorporated into a basement membrane matrix, such as that secreted by Engelbreth-Holm- Swarm mouse sarcoma cells. Such a matrix is sold under the trade name Matrigel™ which resembles the laminin / collagen IV-rich basement membrane extracellular environment found in many tissues. Methods for incorporating agents into such substrates are known in the art.

[0114] The melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may also be moulded into a solid implant, or externally applied patch, suitable for providing efficacious concentrations of the composition over a prolonged period of time without the need for frequent re-dosing. Such controlled release films are well known in the art. Other examples of polymers commonly employed for this purpose that may be used include nondegradable ethylene-vinyl acetate copolymer or degradable lactic acid-glycolic acid copolymers which may be used externally or internally. Certain hydrogels such as poly(hydroxyethylmethacrylate) or poly(vinylalcohol) also may be useful, but for shorter release cycles than the other polymer release systems, such as those mentioned above.

[0115] The carrier may also be a solid biodegradable polymer or mixture of biodegradablepolymers with appropriate time release characteristics and release kinetics. The melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) or a pharmaceutical composition comprising said agent, may then be moulded into a solid implant suitable for providing efficacious concentrations of the active agent over a prolonged period of time without the need for frequent re-dosing. These can be incorporated into the biodegradable polymer or polymer mixture in any suitable manner known to one of skill in the art and may form a homogeneous matrix with the biodegradable polymer, or may be encapsulated in some way within the polymer, or may be moulded into a solid implant.

[0116] The treatment of lung cancer in a subject in the various forms of the present invention may also include treatment of the subject with one or more other interventions, including for example chemotherapy, radiation therapy, immunotherapeutic agent treatment, cancer cell- specific inhibitor treatment, and / or surgery (such as surgical resection of the primary or secondary tumours). In some embodiments, one or more of these other interventions are conducted prior to treatment of the subject with the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). In some embodiments, one or more of these other interventions are conducted concurrently with treatment of the subject with the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). In some embodiments, one or more of these other interventions are conducted after treatment of the subject with the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof).

[0117] Accordingly, in some embodiments the method includes the further step of administering to the subject a chemotherapeutic agent before, during or after administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). Examples of chemotherapeutic agents would be known in the art. For example, suitable chemotherapeutic agents could include etoposide (E; Toposar, Vepesid) along with a platinum (P) agent like cisplatin (Platinol) or carboplatin (Paraplatin) for Small Cell Lung Cancer (SCLC), often called the EP regimen. SCLC in extensive-stages, may also have the EP regimen, combined carboplatin and irinotecan (Camptosar). Up to 90% of people with lung cancer have non-small-cell lung cancer (NSCLC) and examples of chemotherapy treatments for NSCLC include a combination of cisplatin and carboplatin plus either docetaxel (Taxotere), gemcitabine (Gemzar), paclitaxel (Taxol), pemetrexed (Alimta), or vinorelbine (Navelbine). The dosages and dosing regimens contemplated would ultimately be dictated by a skilled physician.

[0118] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) may be administered to the human subject two or more weeks prior to an administration of a chemotherapeutic agent. This is done so that an induction of an immune response in the patient may be achieved prior to the reduction in leukocytes due to treatment with the chemotherapeutic agent.

[0119] In some embodiments the method includes the further step of administering to the subject one or more immunotherapeutic agents before, during or after administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). As would be appreciated by a person skilled in the art, the term "immunotherapeutic agent" as used herein refers to any therapeutic approaches aimed at mobilising or manipulating a subject’s immune system to treat or cure cancer. It includes targeting tumour cells by recognizing the immunogenic proteins or antigens expressed by said tumour cells, which can be accomplished by utilising either passively transferred immune molecules such as antibodies, or preparations designed to induce antibodies or T lymphocytes (T cells) recognising a localised region of an antigen or an epitope specific to the tumor cell. Examples of immunotherapeutic agents may therefore include, but are not limited to, immune checkpoint inhibitors, immunomodulators, activation immunotherapies, suppression immunotherapies, CAR T-cell therapies, and the like.

[0120] Immunomodulators may include interleukins (e.g. IL-2, IL-7, IL-12, IL-15, IL-17 and IL- 21), cytokines (e.g. interferons, and G-CSF), chemokines (e.g. CCL3, CCL26 and CXCL7), immunomodulatory imide agents (IMiDs), and others such as cytosine phosphate-guanosine, oligodeoxynucleotides, and glucans. Other immunomodulators are contemplated. Activation immunotherapies may include dendritic cell-based pump-priming or vaccination, and T-cell adoptive transfer, while suppression immunotherapies may include the use of immunosuppressive drugs and immune tolerance therapries..

[0121] Immune checkpoints are a normal part of the immune system. Generally, their role is to prevent an immune response from being too strong and destroying healthy cells in the body. As part of an effort to harness a cancer patient's own immune system to fight his / her cancer, immunotherapy drugs that belong to a category of immune checkpoint inhibitors have been developed. Those drugs generally work by blocking immune checkpoint proteins, therefore preventing an inhibition of the immune response to cancer cells. For example, administration of immune checkpoint inhibitors may allow T cells to kill cancer cells. Inhibitionof immune checkpoints in cancer patients may also result in inducing the cancer patient's B cells to generate antibodies against the cancer cells.

[0122] The term "immune checkpoint protein" is known in the art. Within the known meaning of this term it will be clear to the person skilled in the art that on the level of "immune checkpoint proteins" the immune system provides inhibitory signals to its components in order to balance immune reactions. Known immune checkpoint proteins may comprise PD1 (also known as PD-1 and Programmed Death 1 receptor) and its ligands PD-L1 and PD-L2, CTLA-4 (Cytotoxic T-lymphocyte-Associated protein, CD152), LAG-3 (Lymphocyte Activation Gene-3), 0X40, A2AR (Adenosine A2A Receptor), BTLA (B and T Lymphocyte Attenuator, CD272), B7-H3 (CD276), B7-H4 (VTCN1), IDO (Indoleamine 2,3-dioxygenase), TIM3 (T-cell Immunoglobulin domain and Mucin domain 3), KIR (Killer-cell Immunoglobulin-like Receptor), VISTA (V-domain Ig Suppressor of T cell Activation), and IL-2R (lnterleukin-2 Receptor). The pathways involving LAGS, BTLA, B7H3, B7H4, TIM3, and KIR are recognized in the art to constitute immune checkpoint pathways similar to the CTLA-4 and PD-1 dependent pathways (see e.g. Pardoll, 2012, Nature Rev. Cancer, 12: 252-264; and Mellman et al., 2011, Nature, 480: 480-489).

[0123] Accordingly, as used herein, an "immune checkpoint inhibitor" is any agent inhibiting the function of an immune checkpoint protein. Inhibition includes reduction of function and full blockade. The designation "immune checkpoint" includes the experimental demonstration of stimulation of an antigen-receptor triggered T lymphocyte response by inhibition of the immune checkpoint protein in vitro or in vivo, e.g. mice deficient in expression of the immune checkpoint protein demonstrate enhanced antigen-specific T lymphocyte responses or signs of autoimmunity (such as disclosed in Waterhouse et al., 1995, Science, 270: 985-988; Nishimura et al., 1999, Immunity, 11: 141-151). It may also include demonstration of inhibition of antigen-receptor triggered CD4+ or CD8+ T cell responses due to deliberate stimulation of the immune checkpoint protein in vitro or in vivo (e.g. Zhu et al., 2005, Nature Immunol. 6: 1245-1252).

[0124] Examples of immune checkpoint inhibitors are antibodies that specifically recognize immune checkpoint proteins. A number of PD1, PD-L1 , PD-L2, CTLA-4, LAG-3, 0X40, A2AR, BTLA, B7-H3, B7-H4, IDO, TIM3, KIR, and IL-2R inhibitors are known and in analogy of these known immune checkpoint inhibitors, alternative immune checkpoint inhibitors may be developed in the future.

[0125] Examples of PD-1 inhibitors include humanized antibodies blocking human PD-1, including but not limited to Pembrolizumab (Keytruda) (e.g. disclosed as hPD109A and its humanized derivatives h409A11, h409A16 and h409A17 in WO2008 / 156712; Hamid et al., N. Engl. J. Med., 369: 134-144 2013), Cemiplimab (Libtayo), Pidilizumab (CureTech) (disclosed in Rosenblatt et al., 2011, J. Immunother., 34: 409-18), and MEDI-0680 (AstraZeneca, previously known as AMP-514).

[0126] PD-1 inhibitors may also include fully human antibodies including but not limited to Nivolumab (Opdivo) (previously known as MDX-1106 or BMS-936558, Topalian et al., 2012, N. Eng. J. Med., 366: 2443-2454, disclosed in US Patent 8,008,449). Other PD-1 inhibitors may include presentations of soluble PD-1 ligand including without limitation PD-L2 Fc fusion protein also known as B7-DC-lg or AMP-244 (disclosed in Mkrtichyan M, et al. 2012, J. Immunol., 189: 2338-2347) and other PD-1 inhibitors presently under investigation and / or development for use in therapy.

[0127] Examples of PD-1L inhibitors may include without limitation humanized or fully human antibodies such as Atezolizumab (Tecentriq) (MPDL3280A), Ave I u ma b (Bavencio) (MSB0010718C), Durvalumab (Imfinzi; MEDI-4736), BMS-936559 / MDX-1105 (Bristol-Myers Squibb), and other PD-L1 inhibitors presently under investigation.

[0128] Examples of CTLA-4 inhibitors include, but are not limited to, Tremelimumab (AstraZeneca; referenced in Ribas et al., 2013, J. Clin. Oncol. 31: 616-22), and Ipilimumab (Yervoy; Bristol-Myers Squibb).

[0129] An Example of a LAG-3 inhibitor includes, but is not limited to, Relatlimab (Bristol- Myers Squibb; BMS-986016). An Example of a B7-H3 inhibitor includes, but is not limited to, MGA271 (Macrogenics). An Example of a KIR inhibitor includes, but is not limited to, Lirilumab (Bristol-Myers Squibb; IPH2102). An Example of an 0X40 inhibitor includes, but is not limited to, MEDI-6469 (Medlmmune). An Example of an IL-2R inhibitor, for preferentially depleting Treg cells (e.g., FoxP-3+ CD4+ cells), comprises IL- 2-toxin fusion proteins, which include, but are not limited to, denileukin diftitox (Ontak; Eisai).

[0130] Accordingly, in some embodiments, the immune checkpoint inhibitor is selected from one or more of the group consisting of a PD-1 inhibitor, a PD-L1 inhibitor, a PD-L2 inhibitor, a CTLA-4 inhibitor, a LAG-3 inhibitor, an 0X40 inhibitor, an A2AR inhibitor, a BTLA inhibitor, a B7-H3 inhibitor, a B7-H4 inhibitor, an IDO inhibitor, a TIM3 inhibitor, a KIR inhibitor, and anIL-2R inhibitor. Examples of such inhibitors are described above. Known inhibitors of these immune checkpoint proteins may be used as such or analogues may be used, in particular chimerized, humanized or human forms of antibodies, as described above.

[0131] In some embodiments, the immune checkpoint inhibitor is selected from one or more of Atezolizumab, Cemiplimab, Durvalumab, Ipilimumab, Nivolumab, and Pembrolizumab.

[0132] As the person skilled in the art will know, alternative and / or equivalent names may be in use for certain antibodies mentioned above. Such alternative and / or equivalent names are interchangeable in the context of the present invention. For example, it is known that Pembrolizumab is also known under the alternative and equivalent names MK-3475 and Lambrolizumab.

[0133] In some embodiments, the one or more immunotherapeutic agents are administered to the subject about one week to about six months or more before or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. For example, the one or more immunotherapeutic agents are administered to the subject about 1 , 2, or 3 weeks, or about 1 , 2, 3, 4, or 5 months before or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0134] In some embodiments the method includes the further step of administering to the subject one or more cancer cell-specific inhibitors before, during or after administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). As used herein, a cancer cell-specific inhibitor refers to a molecule that can specifically target cancer cells but spare normal (non-cancerous) cells. Cancer cell-specific inhibitors can therefore be distinguished from chemotherapeutic agents given that chemotherapeutic agents are unable to distinguish between cancer cells and normal cells, resulting in significant toxicity and side effects to the subject being treated. Cancer cell-specific inhibitors can be broadly classified into two categories, namely small molecules and macromolecules (e.g. monoclonal antibodies, polypeptides, antibody-drug conjugates, and nucleic acids). Accordingly, in some embodiments the cancer cell-specific inhibitor is a small molecule.

[0135] Examples of small molecule cancer cell-specific inhibitors include, but are not limited to, kinase inhibitors (such as receptor tyrosine kinase inhibitors, non-receptor tyrosine kinaseinhibitors, and serineAhreonine kinase inhibitors), epigenetic inhibitors, hedgehog pathway inhibitors, proteasome inhibitors, and poly (ADP-ribose) polymerase (PARR) inhibitors.

[0136] Protein kinases are a type of enzyme that catalyze the transfer of y-phosphate groups from ATP to protein residues containing hydroxyl groups. They have an important role in cell growth, proliferation, and differentiation. The human kinome comprises ~535 protein kinases which can be classified as tyrosine kinases (including both receptor and non-receptor tyrosine kinases), serineAhreonine kinases, and tyrosine kinase-like enzymes, according to their substrate residues.

[0137] Examples of receptor tyrosine kinase inhibitors include anaplastic lymphoma kinase (ALK) inhibitors, cellular-mesenchymal-epithelial transition factor (c-Met) inhibitors, c-Kit (CD117) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, Fms-like tyrosine kinase 3 (FLT3) inhibitors, vascular endothelial growth factor receptor (VEGFR) inhibitors, fibroblast growth factor receptor (FGFR) inhibitors, platelet-derived growth factor receptor (PDGFR) inhibitors, and tropomyosin receptor kinase (TRK) inhibitors. In some embodiments, a receptor tyrosine kinase inhibitor may be selected from the group consisting of crizotinib, certtinib, alectinib, brigatinib, loriatinib, capmatinib, tepotinib, gefitinib, erlotinib, lapatinib, icotinib, afatinib, osimertinib, neratinib, dacomitinib, almonertinib, tucatinib, midostaurin, gilteritinib, quizartinib, pexkiartinib, sorafenib, sunitinib, pazopanib, vandetanib, axitinib, cabozantinib, regorafenib, apatinib, Lenvatinib, tivozanib, fruquintinib, nintedanib, anlotinib, erdafitinib, pemigatinib, avapritinib, ripretinib, selpercatinib, pralsetinib, Larotrectinib and entrectinib. Other receptor tyrosine kinase inhibitors are contemplated.

[0138] Examples of non-receptor tyrosine kinase inhibitors include Bcr-Abl1 inhibitors, Bruton’s agammglobulinemia tyrosine kinase (BTK) inhibitors, and janus kinase (JAK) inhibitors. In some embodiments, a non-receptor tyrosine kinase inhibitor may be selected from the group consisting of imatinib, dasatinib, nilotinib, bosutinib, radotinib, ponatinib, ibrutinib, acalabrutinib, zanubrutinib, ruxolitinib, and fedratinib. Other non-receptor tyrosine kinase inhibitors are contemplated.

[0139] Examples of serineAhreonine kinase inhibitors include BRAF inhibitors, MEK inhibitors, ERK inhibitors, cyclin-dependent kinase (CDK) inhibitors, phosphatidylinositol 3- kinase (PI3K) inhibitors, AKT inhibitors, and mammalian target of rapamycin (mTOR) inhibitors. In some embodiments, a serineAhreonine kinase inhibitor may be selected from the group consisting of dabrafenib, tram etin ib, vemurafenib, encorafenib, cobimetinib,binimetinib, selumetinib, Palbociclib, ribociclib, abemaciclib, idelalisib, copanlisib, duvelisib, alpelisib, temsirolimus, everolimus, and sirolimus. Other serineAhreonine kinase inhibitors are contemplated.

[0140] Examples of epigenetic inhibitors include enhancer of zeste homolog 2 (EZH2) inhibitors, histone deacetylase (HDAC) inhibitors, and isocitrate dehydrogenase 1 / 2 (IDH1 / 2) inhibitors. In some embodiments, an epigenetic inhibitor may be selected from the group consisting of tazemetostat, vorinostat, romidepsin, belinostat, tucidinostat, panobinostat, enasidenib, and ivosidenib. Other epigenetic inhibitors are contemplated.

[0141] Examples of hedgehog pathway inhibitors include venetoclax, vismodegib, sonidegib, glasdegib, bortezomib, carfilzomib, ixazomib, olaparib, rucaparib, niraparib and talazoparib. Other hedgehog pathway inhibitors are contemplated.

[0142] Examples of proteasome inhibitors include bortezomib, carfitzomib, ixazomib, marizomib, oprozomib, and delanzomib. Other proteasome inhibitors are contemplated.

[0143] Examples of PARR inhibitors include Nicotinamide, Olaparib, rucaparib, niraparib, talazoparib, pamiparib, veliparib, iniparib, amelparib, and fluzoparib. Other PARR inhibitors are contemplated.

[0144] In some embodiments, the one or more cancer cell-specific inhibitors are administered to the subject about one week to about six months or more before or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. For example, the one or more cancer cell-specific inhibitors are administered to the subject about 1 , 2, or 3 weeks, or about 1 , 2, 3, 4, or 5 months before or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0145] The dosage and frequency of dose of the immunotherapeutic agent(s) and / or cancer cell-specific inhibitors) administered to the subject before, during or after treatment with the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) may vary depending on the half-life of the agent or inhibitor in the subject. It would be understood by a person skilled in the art that such guidelines would be adjusted for the molecular weight of the agent or inhibitor, the clearance from the blood, the mode of administration, and other pharmacokinetic parameters. The dosage may also be varied forlocalized administration, e.g. intranasal, inhalation, subcutaneous, etc., or for systemic administration.

[0146] In some embodiments, the immunotherapeutic agent(s) and / or the cancer cell-specific inhibitors) may be administered to the subject over one or more cycles coordinated with the cyclic administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) as described above. For example, the subject may receive a dose of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), then a dose of one or more immunotherapeutic agents and / or one or more cancer cell-specific inhibitors, then a further dose of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), etc, repeated in a cyclic manner. In this instance, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), the immunotherapeutic agent(s) and / or the cancer cell-specific inhibitors), are administered separately and at discreet time points from each other.

[0147] It will be appreciated that the immunotherapeutic agent(s) and / or cancer cell-specific inhibitors) are administered to the subject in an effective amount. The term "effective amount" in this regard has been described above with respect to the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). For example, particular characteristics dictating an effective amount of the immunotherapeutic agent(s) and / or cancer cell-specific inhibitors) to be administered include (but are not limited to) the mode of administration, the chemical nature, structure and size of the agents or inhibitor, relevant pharmacokinetic properties of the agent or inhibitor, and the characteristics of the subjectas indicated above. A person skilled in the art will be able to determine appropriate dosages for the immunotherapeutic agent(s) and / or cancer cell-specific inhibitors) depending on these and other factors.

[0148] The manner in which the chemotherapeutic agent, the immune immunotherapeutic agent(s), and / or the cancer cell-specific inhibitors), are administered to the subject may be selected from those described above with respect to the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof).

[0149] In some embodiments, the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) may be administered to the subject about 1 to 2, or about 2 or more, weeks prior to an administration of a chemotherapeutic agent and / or theimmunotherapeutic agent(s) and / or the cancer cell-specific inhibitors). This is done so that an induction of an immune response in the patient is achieved.

[0150] In some embodiments, the method of the present invention includes the further step of one or more administrations of an effective amount of an agent to the subject that reduces the level and / or activity of regulatory T cells in the subject. Regulatory T cells are CD4+CD25+ T cells (reviewed in Terabe and Berzofsky, 2004, Curr. Opin. Immunol., 16(2): 157-162). Methods for assessing the level and / or activity of regulatory T cells are known in the art.

[0151] In the case where surgical interventions are used, the treatment method of the present invention may include the further step of resecting metastases from the subject. In some embodiments, the metastases are resected prior to administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). In some embodiments, the metastases are resected after administration of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). Metastatic involvement of organs may include skin, subcutaneous tissues, muscle, lung, liver, brain, bowel, spleen and bone.

[0152] The method of treating the subject in the various forms of the present invention may also include the step of selecting a subject suitable for treatment. In one form, the subject is selected for treatment on the basis that the subject shows a T cell response to a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof).

[0153] Methods for assessing whether a subject shows a T cell response to a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, are known in the art. An antigen of the cell lysate will be understood to mean an antigen normally present in the melanoma cell lysate that is involved in the induction of an immune response to a melanoma in the subject.

[0154] Accordingly, in one aspect the present invention provides a method of selecting a subject with lung cancer suitable for treatment with the method of treatment as described herein, the method including the step of identifying a subject that shows a T cell response upon administration to the subject of a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0155] The subject may also be selected on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administering to the subject a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof).

[0156] Accordingly, in one aspect the present invention provides a method of selecting a subject with lung cancer suitable for treatment with the method of treatment as described herein, the method including the step of identifying a subject that has a decreased level and / or activity of regulatory T cells upon administration to the subject of a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. As discussed above, regulatory T cells are CD4+CD25+ T cells and methods for assessing the level and / or activity of regulatory T cells are known in the art.

[0157] As indicated above, the present invention may be used to inhibit, prevent and / or slow the formation and / or growth of secondary tumours in a subject. Therefore, the present invention may also be used to inhibit or prevent metastases in the subject. The extent of formation and / or growth of primary and secondary tumours in a subject may be monitored by a method known in the art. Monitoring methods include clinical examination and measurement, U / S scans, CT scans, chest X ray, bone X ray, MRI scans, PET scans, and bone scans.

[0158] Accordingly, in one aspect the present invention provides a method of inhibiting formation and / or growth of a secondary tumour in a subject with lung cancer, the method including the step administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0159] The present invention may also be used to inhibit and / or prevent the growth of the primary tumour in a subject, including reducing the growth of the primary tumour after resection or other treatment.

[0160] Accordingly, in one aspect the present invention provides a method of inhibiting growth of a primary lung cancer in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0161] The present invention may also be used to improve the rate of survival of a subject with a lung cancer. An improved rate of survival can be measured, for example, as animproved survival rate after a given amount of time (e.g. 2 to 10 years or longer depending on the stage of the cancer), or an improved median survival time.

[0162] Accordingly, in one aspect the present invention provides a method of improving the rate of survival of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof. Methods for determining the improvement in survival rate are known in the art.

[0163] The present invention may also be used to improve the life expectancy of a subject. Accordingly, in one aspect the present invention provides a method of increasing life expectancy of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0164] The present invention may also be used to improve the outcome for a subject suffering from lung cancer, and / or improve the quality of life of the subject. Methods for assessing outcome and quality of life parameters are known in the art.

[0166] The present invention may also be used to induce an anti-tumour immune response in a subject. Accordingly, in one aspect the present invention provides a method of inducing an anti-tumour immune response in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

[0166] Methods for assessing an anti-tumour response in a subject are known in the art.

[0167] Induction of anti-lung cancer immunity may be determining by a delayed type hypersensitivity (DTH) response against lung cancer antigens prior to and at periods after the melanoma cell lysate treatment. In addition, serum and peripheral blood lymphocytes may be obtained prior to injection of the melanoma cell lysate and at periods after melanoma cell lysate administration to test the induction of anti-lung cancer immunity by cytotoxicity assay, CTL precursor frequency (CTLp) assay and phenotypic analysis of lymphocytes.

[0168] In some embodiments of the aforementioned aspects of the present invention, the subject is a human subject. In some embodiments, the melanoma cell lysate includesfragmented melanoma cell membranes. In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the viral melanoma cell lysate is a vaccinia melanoma cell lysate.

[0169] In some embodiments of the aforementioned aspects of the present invention, the treatment method is capable of producing one or more of an increase in the number of intratumoral dendritic cells in the subject, an improvement in dendritic cell activation in the subject, and a modulation (i.e. increase or decrease) in the number and / or activity of regulatory T cells in the subject. Assays to determine these treatment outcomes would be known in the art.

[0170] The terms "increase" and "improve" and the like as used throughout the specification are to be understood to mean a greater level in the progress of a process, including any one or more of the start, continuation or termination of a process. For example, the terms "increase", and "improve" can refer to an at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater, or by 1-fold, 2-fold, 3-fbld, 4-fold, 5-fold, 6.0-fold, 7-fold, 8-fbld, 9-fold, 10-foki, 15-foki, 20-fbki, 25-foki, 30-foki, 35-foki, 40-fold, 45-fold, 50-fold, 60- fold, 70-fbld, 80-fold, 90-fold, 100-foki, 125-foki, 150-foki, 175-fold, 200-fbki, 225-foki, 250- fold, 275-fbki, 300-fold, 400-fold, 500-foki, 1000-fbki, 10,000-foki, 100,000-fold, or greater, level when compared to the level of intratumoral dendritic cells or dendritic cell activation in a subject which has not been treated using the method of the present invention, or when compared to the number and / or activity of regulatory T cells in a subject which has not been treated using the method of the present invention.

[0171] Similarly, the term "decrease" refers to an at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or greater, or by 1-fold, 2-fold, 3-fbld, 4-fold, 5-fold, 6.0- fold, 7-fold, 8-fold, 9-fold, 10-foki, 15-foki, 20-fbld, 25-fold, 30-fbld, 35-fold, 40-foki, 45-fold, 50-foki, 60-fbld, 70-fold, 80-foki, 90-foki, 100-foki, 125-foki, 150-fold, 175-fold, 200-foki, 225- fold, 250-foki, 275-fbld, 300-fold, 400-foki, 500-foki, 1000-foki, 10,000-foki, 100,000-foki, or less, number and / or activity when compared to the number and / or activity of regulatory T cells in a subject which has not been treated using the method of the present invention.

[0172] The present invention also provides use of a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) in the preparation of a medicament for treating lung cancer in a subject.

[0173] In some embodiments, the present invention provides use of a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) in the preparation of a medicament for treating lung cancer in a human subject.

[0174] In some embodiments, the melanoma cell lysate includes fragmented cell membranes. Accordingly, the present invention also provides use of a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) in the preparation of a medicament for treating lung cancer, wherein the melanoma cell lysate includes fragmented melanoma cell membranes.

[0175] In some embodiments, the melanoma cell lysate is a viral melanoma cell lysate. In some embodiments, the melanoma cell lysate is a vaccinia melanoma cell lysate. Accordingly, the present invention also provides the use of a vaccinia melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) in the preparation of a medicament for treating lung cancer.

[0176] In a further aspect, the present invention provides a pharmaceutical composition when used for treating lung cancer in a subject, the composition including a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof.

[0177] In a further aspect, the present invention provides a combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and (ii) a chemotherapeutic agent; and / or (ill) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

[0178] In a further aspect, the present invention provides a combination product when used for treating or preventing lung cancer in a subject, the combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and(ii) a chemotherapeutic agent; and / or(iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

[0179] In some embodiments, the components of the combination product are provided in a form for separate administration to the subject.

[0180] In the context of the present invention, the "combination product" may be in the form of a kit which includes each component.

[0181] In some embodiments, the composition or combination product is used to treat a human subject with lung cancer.

[0182] In some embodiments, the composition or combination product inhibits growth of a primary lung cancer in the subject, inhibits formation and / or growth of a secondary tumour in the subject, improves rate of survival of the subject, improves life expectancy of the subject, and / or induces an anti-tumour immune response in the subject.

[0183] Accordingly, in further aspects the present invention provides a combination product for use in any one or more of: treating or preventing lung cancer in a subject; inhibiting growth of a primary lung cancer in a subject; inhibiting formation and / or growth of a secondary tumour in a subject; improving the rate of survival of a subject with lung cancer; improving the life expectancy of a subject with lung cancer; and inducing an anti-tumour immune response in a subject with lung cancer, wherein the combination product includes the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and(ii) a chemotherapeutic agent; and / or(iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

[0184] In further aspects, the present invention provides use of a combination product for any one or more of: treating or preventing lung cancer in a subject; inhibiting growth of aprimary lung cancer in a subject; inhibiting formation and / or growth of a secondary tumour in a subject; improving the rate of survival of a subject with lung cancer; improving the life expectancy of a subject with lung cancer; and inducing an anti-tumour immune response in a subject with lung cancer, wherein the combination product includes the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and(ii) a chemotherapeutic agent; and / or(iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors..

[0186] The combination product may also include instructions on when and how to administer each component of the combination product to the subject. The combination product may also include instructions for selecting a subject with lung cancer suitable for treatment using the combination product. Such instructions may, for example, include instructions for selecting a subject with lung cancer suitable for treatment on the basis that the subject shows a T cell response to a melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof), and / or for selecting a subject on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administration to the subject of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof). In these instances, the combination product may be supplied in the form of a kit for performing the relevant function.

[0186] Methods for packaging the various components of the combination product are known in the art.

[0187] The nature of the melanoma cell lysate (and / or an immunotherapeutic extract, component or antigen thereof) and the agents and / or inhibitors to be included in the composition or combination product have been described in detail above. In some embodiments, the one or more immunotherapeutic agents are one or more immune checkpoint inhibitors, as described above.General Methods

[0188] Activation, phenotype and function of T cells may be tested through ELISPOT, cytokine assays, cytotoxicity assays and proliferation assays.

[0189] IFN-y and IL4 ELISPOT, antigen specific3H-thymidine incorporation proliferation assay and flow cytometry may be used to analyse regulatory T cells.

[0190] For T cell assays, PBMC will be isolated from peripheral blood using standard Ficoll- Hypaque density centrifugation methods. PBMC may be frozen in aliquots suitable for the experiments and thawed when required. Blood from healthy or other volunteers may be used as controls to establish the ‘normal’ range of responses and background values for the various tests used.

[0191] IFN-y and IL4 ELISPOT assays may be used to test for the frequency of tumour antigen reactive T cells (CTL and T helper cells). Commercially available antibody pairs (R&D systems for IFN-y and BD Pharmingen for IL4) may be used in a standard ELISPOT protocol. PBMC may be cultured with various antigens in 96-well PVDF membrane microwell plates (Millipore), which have been coated before with the appropriate amount of antibody. Antigens that may be used include vaccinia melanoma cell lysate, MM200 lysate, autologous tumour cell lysate (were applicable), Tet-Tox as a recall antigen and PHA as a positive control. Peripheral blood mononuclear cells (PBMCs) without adding any antigen may be used as a background control.

[0192] Antigen specific proliferation of PBMCs may be tested using standard3H-thymidine incorporation assays. PBMCs are cultured in 96 well plates in the presence of antigen for 2 to 3 days.3H thymidine may be added for the last 16h of the culture.

[0193] The Miltenyi Cytokine Secretion Assay may be used to detect IFN-y and IL4 producing T cells in the peripheral blood. This is a very sensitive assay giving information on the frequency of cytokine producing cells. Additionally, cells can be counterstained with cell surface markers to identify the subpopulation of cells. CD3 may be used as a general T cell marker and CD8 to distinguish between CD8 and CD4 T cells as cell surface markers. PBMCs cultured overnight (16h) in the presence of vaccinia melanoma cell lysate, MM200 cell lysate or autologous tumour cell lysate (where applicable) may be used in this assay.

[0194] Cytotoxicity of activated T cells may be tested with standard calcein-Acetoxymethyl (AM) (Molecular Probes) cytotoxicity assays (Lichtenfels et al., 1994, J. Immunol. Methods, 172(2): 227-239; Neri et al., 2001 , Clin. Diagn. Lab. Immunol, 8(6): 1131-1135). This a nonradioactive, very sensitive cytotoxicity assay with comparable results to the chromium release assay. Calcein-AM freely diffuses into most cells. Once inside the cell, this nonfluorescent substrate is converted by nonspecific intracellular esterases into a fluorescent product that are retained by cells with intact plasma membranes. In contrast, the dye will leak from cells with damaged cell membranes and can be detected in the supernatant.

[0195] PBMCs may be tested for cytotoxicity towards calcein-AM labelled cells either directly after being isolated from the peripheral blood (fresh) or after being cultured in vitro for one week with antigen. Target cells used in this assay would be autologous tumour cells (where applicable), MM200 cells or other HLA-matched melanoma cell lines. Autologous monocyte derived DC (MoDC) pulsed or unpulsed with tumour cell lysate as target cells may be used if no suitable HLA-matched melanoma cell line can be used. MoDC may be generated using standard protocols with IL4 and GMCSF as described in (Heinzel et al., 2001, Cancer Immunol. Immunother., 49(12): 671-8). Target cells may be labelled with calcein-AM for 30 min at 37°C. 104target cells may be incubated with effector cells (50:1 to 1:1 ratios) for 4 hours. Supernatants may then be transferred to new wells and measured using a Typhoon 9410 (Amersham Biosciences). (Excitation 488nm, emission 530nm). Percent lysis is calculated with the same standard formula used for chromium release assays.

[0196] Flow cytometry may be used to investigate the phenotype of activated T cells. Fresh PBMCs, or after 10-day culture in the presence of antigen, can be analysed for the expression of markers including CD3, CD4, CD8, CD25, CD16, CD56, CD69, CD45RO.

[0197] To test for the cytokine profile of activated T cells, PBMCs can be stimulated in vitro in the presence of antigen (MoDC pulsed with MM200 lysate or vaccinia melanoma cell lysate). After 10 days of culture, cells can be tested with intracellular cytokine staining and flow cytometric analysis for the production of cytokines. Cells may be harvested and stimulated for 4 hours with PHA / lonomycin in the presence of Brefeldin A. Cytokines suitable for testing include 112, IL4, IL10, IL13, TNFa and IFN-g. Cells may also be stained with cell surface markers and analysed using three colour flow cytometry. Cell surface markers include CD3, CD4, CD8, CD69, CD25 and CD45RO.

[0198] The presence of anti-ganglioside antibodies in the sera from subjects may be tested. Antiganglioside antibodies may be detected by immunodot-blot. Strips of PVDF-P membranes (Millipore) are coated with purified commercial GM3, GM2, GD3, GM1, GD1a, GD1b, and GT1b gangliosides (Sigma). The strips are incubated with 1 / 100, 1 / 200, and 1 / 500 dilutions of the patients’ sera. Bound antibodies can be detected with alkaline phosphatase conjugated antibodies to human IgG and IgM. BCIP / NBT will be used as substrate.

[0199] PBMCs activated in vitro with vaccinia melanoma cell lysate and tumour cell lysate may be tested to determine whether a vaccinia melanoma cell lysate or tumour specific response can be induced and to test whether the type of T cell activated by vaccinia melanoma cell lysate is comparable in vitro and in vivo. Activated T cells may be phenotyped as described above. PBMC can be stimulated in vitro with autologous MoDC pulsed with vaccinia melanoma cell lysate, MM200 cell lysate, irradiated MM200 cells, and autologous tumour cell lysate (where applicable). Tet-Tox can be used as a control. After 10 to 14 days of culture T cells may be restimulated once with antigen pulsed MoDC and cultured for another 10-14 days. IL2 may be added to the culture 2 days after restimulation. General activation can be tested with3H-thymidine proliferation assay. Frequency of vaccinia melanoma cell lysate or MM200 reactive T cells may be measured with ELISPOT and Miltenyi cytokine secretion assay.

[0200] The relative number of CD4+CD25+ Treg cells in the peripheral blood of patients may be determined. This is done by comparing the number of Tregs in patients before and after vaccination; in addition it is also possible to compare the number of CD4+CD25+ Treg cells between different patient groups, namely responders to the vaccine with non-responders, and Stage IV to Stage III patients. The frequency of CD4+CD25 Treg cells can be determined using 3 colour flow cytometry. Cells may be stained with CD4 and CD25 antibodies plus other important markers to define the population as Treg cells. Suitable markers are CD45RA, CD45RO, CD62L, CD122. The population of interest is the CD4+, CD25high population. Most of these cells also express CD45RO, CD62L and CD122 and will not express CD45RA. The Treg (CD4*CD25*) fraction may be assessed in association with the induction of specific CD8* cells.

[0201] To test the functionality of the CD4*CD25high cell population to inhibit normal T cell responses, the CSFE cell division assay may be used (Lyons and Parish, 1994, Immunol. Methods 171(1): 131-7). T cells from peripheral blood can be separated into CD8* and CD4+fractions. The CD4+ cell may be further separated into the CD25neg and the CD25high population. The CD25neg population can be stained with CFSE, and activated non- specifically with anti-CD3 and anti-CD28 antibodies or specifically with vaccinia melanoma cell lysate or MM200 tumour cell lysate pulsed autologous MoDC and co-cultured for 4 days with or without the CD25high population. Cell division may be determined in the flow cytometer. CD4*CD25* cells can also be tested for their cytokine production, using intracellular cytokine staining as described above. Cytokines tested for include IL10 and TGF-0.

[0202] Unless otherwise indicated, the practice of many aspects of the present invention employs conventional techniques of molecular biology, recombinant DNA technology and immunology, which are within the skill of the art. Such techniques are described in more detail in the scientific literature, for example, Green MR and Sambrook J, Molecular Cloning: A Laboratory Manual (4th edition), Cold Spring Harbor Laboratory Press, 2012, Ausubel, F. M. et al. Current Protocols in Molecular Biology, Wiley-lnterscience, New York, current volume; Albers, B. et al., Molecular Biology of the Cell, 2.sup.nd Ed., Garland Publishing, Inc., New York, N.Y. (1989); Lewin, B M, Genes IV, Oxford University Press, Oxford, (1990); Watson, J. D. et al., Recombinant DNA, Second Edition, Scientific American Books, New York, 1992; Darnell, J E et al., Molecular Cell Biology, Scientific American Books, Inc., New York, N.Y. (1986); Old, R. W. et al., Principles of Gene Manipulation. An Introduction to Genetic Engineering, 2.sup.nd Ed., University of California Press, Berkeley Calif. (1981); DNA Cloning Practical Approach, vol. I & II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., Current Edition); Nucleic Acid Hybridization (B. Hames & S. Higgins, eds., Current Edition); Transcription and Translation (B. Hames & S. Higgins, eds., Current Edition); Methods in Enzymology: Guide to Molecular Cloning Techniques, (Berger and Kimmel, eds., 1987); Hartlow, E. et al., Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988), Coligan, J. E. et al., eds., Current Protocols in Immunology, Wiley-lnterscience, New York 1991.

[0203] It is to be noted that where a range of values is expressed, it will be clearly understood that this range encompasses the upper and lower limits of the range, and all numerical values or sub-ranges in between these limits as if each numerical value and sub- range is explicitly recited. The statement "about X% to Y%" has the same meaning as "about X% to about Y%," unless indicated otherwise.

[0204] The term "about" as used in the specification means approximately or nearly and in the context of a numerical value or range set forth herein is meant to encompass variations of + / - 10% or less, + / - 5% or less, + / - 1% or less, or + / - 0.1% or less of and from the numerical value or range recited or claimed.

[0205] As used herein, the singular forms "a," "an," and "the" may refer to plural articles unless specifically stated otherwise. Therefore, it is to be made clear that reference to an antimicrobial agent being present in the antimicrobial compositions described herein includes reference to use of a combination of antimicrobial agents in the composition.

[0206] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

[0207] It will be apparent to the person skilled in the art that while the invention has been described in some detail for the purposes of clarity and understanding, various modifications and alterations to the embodiments and methods described herein may be made without departing from the scope of the inventive concept disclosed in this specification.

[0208] The invention is further illustrated in the following examples. The examples are for the purpose of describing particular embodiments only and are not intended to be limiting with respect to the above description.EXAMPLE 1Preparation of Melanoma Cell Lysate

[0209] Melanoma cell lysate preparation was carried out as in Hersey et al., 1987, Cancer Immunol. I mm u noth er. 25: 257-265, using methods similar to those described by Wallack et al, 1998, J. Am. Coll. Surg. 187:69-77 and Wallack et al., 1986, Cancer 57:649-655.

[0210] A single allogeneic melanoma cell line, referred to as MM200 (Pope et al., 1979, Pathology 11 : 191 -195), was infected with vaccinia virus (prepared by Commonwealth Serum Laboratories, Melbourne, Australia) at 2.5 pock-fbrming unrts / cell. MM200 was originally isolated from a primary melanoma on a 43-year-old woman in 1972 but no other patient details are available. The HLA type was A1 ,3; B7.35; DR2.4. Chromosomal analysis showeda nodal number of 76, and a number of marker chromosomes were revealed by karyotypic analysis (Muir and Gunz, 1979, Pathology 11:597-606).

[0211] The vaccinia virus was the strain referred to as strain O, which was derived in 1921 from two strains imported from the Lister Institute in 1912 and a strain from Japan imported in 1913. After 24 hours of incubation, the lysed cells were further homogenized with a sterile Dounce (type B pestle) homogenizer and centrifuged at 400 x g for 7 minutes. The supernatant (1) was kept and the pellet frozen and thawed in 1 to 3 mL of distilled sterile water. The latter was then made up to 20 mL, centrifuged at 400 x g for 7 minutes and the supernatant (2) added to supernatant (1). The pooled supernatant was centrifuged at 38,000 x g for 60 minutes and the sediment resuspended in saline to give an equivalent of 5 x 106MM200 cells / 0.5 mL saline. The lysate was tested for pathogenic viral, bacterial, or fungal contamination and kept at -80°C until use.EXAMPLE 2VMCL - Stage IV Lung Cancer Case Study 1

[0212] A 73-year-old male patient with Stage IV advanced metastatic lung cancer was treated initially with combination multi-agent chemotherapy from 15 March 2021 to Nov 2021 using Paclitaxel, Carboplatin and anti-PD1 immunotherapy (Pembrolizimab); then radiotherapy to lumbar and cervical spine metastases during December 2021 and January 2022; then from February until April 2022 with Docetaxel and Dostarlimab anti-PD-1 antibody therapy and Cobolimab (TSR-022, a novel lgG4 anti-TIM-3 mAb), as part the ‘COSTAR Lung’ clinical trial. The treatment failed to control tumour growth and he continued to develop progressive metastatic disease.

[0213] The patient was treated initially on 14 / 07 / 2022 with Vaccinia Melanoma Cell Lysate (VMCL) Immunotherapy alone, and then fortnightly 0.3 mis injected intradermally for another two VMCL doses alone. Several of his scalp subcutaneous metastases regressed substantially (> 50% of their initial size). Most notably, one laterally based scalp metastasis completely regressed, so that it was no longer measurable or visible.

[0214] This demonstrates a substantial clinical effect of the VMCL when administered alone and demonstrates its clinical activity in lung cancer. This observation is the first time that the clinical efficacy of a melanoma vaccine, and specifically for VMCL vaccine immunotherapy, has been noted and reported for lung cancer.EXAMPLE 3VMCL - Stage IV Lung Cancer Case Study 2

[0215] A 71 -year-old woman with Stage IV advanced metastatic lung cancer was treated initially with combination multi-agent chemotherapy and anti-PD1 immunotherapy. The treatment failed to control tumour growth and she continued to develop progressive disease. The patient initially developed a mass in the right lower lobe of her lung, which was diagnosed in November 2019 as Stage IV Non-Small Cell Lung Carcinoma (NSCLC). The mass was incidentally detected on CT scanning while being investigated for Colon Cancer. The colon cancer mass was completely resected later in 2019, with multiple repeated normal colonoscopies since, and no evidence of recurrent colon cancer.

[0216] The NSCLC was not able to be resected and so the subject was treated with chemotherapy, radiotherapy and immunotherapy. Durvalumab (anti-PD-1 inhibitory monoclonal antibody therapy) immunotherapy was commenced in March 2022 but was ceased in June 2022 due to serious toxicity after her 4th dose with severe pulmonary complications (pneumonitis and infection). Radiation treatment was given to the right lung and hilar region (weekly for 7 weeks (7 / 2 / 2020 to 20 / 3 / 2020). Chemotherapy was administered comprising Cisplatin days 1-8 and Etoposide days 1-5 repeated each 28 days (monthly) until it was deemed unsuccessful due to development of recurrent biopsy proven malignant pleural effusions which required admission and pleural drainage procedures on several occasions in late 2020. The dosages and dosing regimens used were given in the recommended standard doses and adjusted and calculated according to the patient’s responses and effects by a skilled physician.

[0217] Bronchosopy and biopsies were performed on 23 / 07 / 2020 for an unresolved right lower lobe of lung infiltrate and showed malignant cells consistent with NSCLC. The right hilar / mediastinal mass was biopsied by Endoscopic Bronchial UltraSound (EBUS) on 27 / 11 / 2020 showing malignant metastatic cells and narrowing in right bronchus. Eventually, after several further repeated drainages of the pleural fluid, a talc pleurodesis procedure via chest drain tube was performed on 19 / 10 / 2022 in an effort to reduce fluid collection, during which the malignant nature of the pleural effusion was again confirmed on cytology. It was decided to cease any further chemotherapy or immunotherapy due to failure of the previous treatments and severe side-effects of those treatments.

[0218] The patient had become increasingly unwell from her lung compromise with shortness of breath at rest, a cough, some weight loss and progressive functional deterioration, unableto perform many of her usual daily activities and some self-care activities, requiring assistance from her husband.

[0219] The patient was then treated with Vaccinia Melanoma Cell Lysate (VMCL) Immunotherapy alone. The VMCL was prepared as described above in Example 1. After 4 doses of the VMCL (each dose given about every two weeks) her condition continued to improve with reduction in shortness of breath, maintenance of body weight and increased exercise tolerance where she almost completely returned to her normal daily activities. Most notably, after 3 months of VMCL therapy CT scans showed the stability of her disease and early regressive changes of the lung disease with no new metastases developing, and the malignant pleural effusion not recurring nor requiring any pleural drainage procedures; with her disease remaining stable (CT scans 19 / 8 / 2024), and with survival and improved physical function now for a period of over 20 months so far.

[0220] This demonstrates a substantial clinical effect of the VMCL when administered alone and demonstrates its clinical activity in lung cancer.

[0221] The aforementioned case studies demonstrate the clinical efficacy of a melanoma vaccine, and specifically for VMCL vaccine immunotherapy, for the treatment of lung cancer. These results also show the effectiveness of the treatment protocol in preventing lung cancer recurrence, preventing the growth of a further primary cancer, and preventing the formation and / or growth of one or more secondary tumours in the subject.

[0222] The description provided herein is in relation to several embodiments which may share common characteristics and features. It is to be understood that one or more features of one embodiment may be combinable with one or more features of the other embodiments. In addition, a single feature or combination of features of the embodiments may constitute additional embodiments.

[0223] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to, or indicated in this specification, individually or collectively, and any and all combinations of any two or more of the steps or features.

Claims

CLAIMS1. A method of treating or preventing lung cancer in a subject, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

2. The method according to claim 1 , wherein the subject is a human subject.

3. The method according to claim 1 or claim 2, wherein the melanoma cell lysate includes fragmented melanoma cell membranes.The method according to any one of claims 1 to 3, wherein the melanoma cell lysate is a viral melanoma cell lysate.

5. The method according to claim 4, wherein the viral melanoma cell lysate is a vaccinia melanoma cell lysate.

6. The method according to any one of claims 1 to 3, wherein the melanoma cell lysate is an allogeneic cell lysate.

7. The method according to claim 6, wherein the melanoma cell lysate includes a lysate from MM200 cells.

8. The method according to any one of claims 1 to 7, wherein the lung cancer is Stage IV lung cancer.

9. The method according to claim 8, wherein the Stage IV lung cancer is advanced Stage IV lung cancer.

10. The method according to any one of claims 1 to 9, wherein the administration of the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, to the subject includes multiple administrations of the melanoma cell lysate to the subject during a treatment period.

11. The method according to claim 10, wherein the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for one or more cycles.

12. The method of claim 11, wherein the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for at least about one month.

13. The method of claim 11 or claim 12, wherein the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, is administered to the subject about every two weeks for at least about three months.

14. The method of any one of claims 1 to 13, wherein the subject undergoes radiation therapy and / or chemotherapy and / or surgery before, during or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

15. The method according to any one of claims 1 to 14, wherein the method includes the further step of administering to the subject one or more of: one or more chemotherapeutic agents; one or more immunotherapeutic agents; and one or more cancer cell-specific inhibitors, before, during or after administration of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

16. The method according to claim 15, wherein the one or more immunotherapeutic agents comprise one or more immune checkpoint inhibitors.

17. The method according to any one of claims 1 to 16, wherein the method includes the further step of one or more administrations of an effective amount of an agent to the subject that reduces the level and / or activity of regulatory T cells in the subject.

18. The method according to any one of claims 1 to 17, wherein the method includes the step of selecting a subject suitable for treatment.

19. The method according to claim 18, wherein the subject is selected on the basis that the subject shows a T cell response to a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

20. The method according to claim 18 or claim 19, wherein the subject is selected on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administration to the subject of the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

21. The method according to any one of claims 1 to 20, wherein formation and / or growth of a secondary tumour in the subject is inhibited.

22. The method according to any one of claims 1 to 21 , wherein growth of a primary lung cancer in the subject is inhibited.

23. The method according to any one of claims 1 to 22, wherein rate of survival of the subject is improved.

24. The method according to any one of claims 1 to 23, wherein life expectancy of the subject is improved.

25. The method according to any one of claims 1 to 24, wherein an anti-tumour immune response in the subject is induced.

26. The method according to any one of claims 1 to 25, wherein one or more of an increase in the number of intratumoral dendritic cells is produced in the subject, dendritic cell activation is improved in the subject, and the number and / or activity of regulatory T cells is decreased in the subject.

27. A method of inhibiting formation and / or growth of a secondary tumour in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

28. A method of inhibiting growth of a primary lung cancer in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

29. A method of improving the rate of survival of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

30. A method of increasing life expectancy of a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

31. A method of inducing an anti-tumour immune response in a subject with lung cancer, the method including the step of administering to the subject a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

32. The method according to any one of claims 27 to 31 , wherein the subject is a human.

33. The method according to any one of claims 27 to 32, wherein the melanoma cell lysate includes fragmented melanoma cell membranes.

34. The method according to any one of claims 27 to 33, wherein the melanoma cell lysate is a viral melanoma cell lysate.

35. The method according to claim 34, wherein the melanoma cell lysate is a vaccinia melanoma cell lysate.

36. The method according to any one of claims 27 to 35, wherein the lung cancer is Stage IV lung cancer.

37. Use of a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof, in the preparation of a medicament for treating lung cancer in a subject.

38. The use according to claim 37, wherein the subject is a human.

39. The use according to claim 37 or claim 38, wherein the melanoma cell lysate includes fragmented melanoma cell membranes.

40. The use according to any one of claims 37 to 39, wherein the melanoma cell lysate is a viral melanoma cell lysate.

41. The use according to claim 40, wherein the melanoma cell lysate is a vaccinia melanoma cell lysate.

42. The use of any one of claims 37 to 41 , wherein the lung cancer is Stage IV lung cancer.

43. A pharmaceutical composition when used for treating lung cancer in a subject, the composition including a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof.

44. A combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and00 a chemotherapeutic agent; and / or(iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

45. A combination product when used for treating or preventing lung cancer in a subject, the combination product including the following components:(i) a melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof; and00 a chemotherapeutic agent; and / or (iii) an agent that reduces the level and / or activity of regulatory T cells in the subject to be treated; and / or(iv) one or more immunotherapeutic agents; and / or(v) one or more cancer cell-specific inhibitors.

46. The combination product according to claim 44 or claim 45, wherein the components are provided in a form for separate administration to the subject.

47. The composition or combination product according to any one of claims 43 to 46, wherein the melanoma cell lysate includes fragmented melanoma cell membranes.

48. The composition or combination product according to any one of claims 43 to 47, wherein the melanoma cell lysate is a viral melanoma cell lysate.

49. The composition or combination product according to claim 48, wherein the melanoma cell lysate is a vaccinia melanoma cell lysate.

50. The composition or combination product according to any one of claims 43 to 49, wherein the melanoma cell lysate is an allogeneic cell lysate.

51. The composition or combination product according to claim 50, wherein the melanoma cell lysate includes a lysate from MM200 cells.

52. The combination product according to any one of claims 44 to 51 , wherein the one or more immunotherapeutic agents comprise one or more immune checkpoint inhibitors.

53. The composition or combination product according to any one of claims 43 to 52, wherein the lung cancer is Stage IV lung cancer.

54. The composition or combination product according to any one of claims 43 to 53, wherein the composition or combination product inhibits growth of a primary lung cancer in the subject.

55. The composition or combination product according to any one of claims 43 to 54, wherein the composition or combination product inhibits formation and / or growth of a secondary tumour in the subject.

56. The composition or combination product according to any one of claims 43 to 55, wherein the composition or combination product improves rate of survival of the subject.

57. The composition or combination product according to any one of claims 43 to 56, wherein the composition or combination product improves life expectancy of the subject.

58. The composition or combination product according to any one of claims 43 to 57, wherein the composition or combination product induces an anti-tumour immune response in the subject.

59. The combination product according to any one of claims 44 to 58, wherein the combination product includes instructions for selecting a subject with lung cancer suitable for treatment.

60. The combination product according to claim 59, wherein the product includes instructions for selecting a subject with lung cancer suitable for treatment on the basis thatthe subject shows a T cell response to the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

61. The combination product according to claim 59 or claim 60, wherein the product includes instructions for selecting a subject with lung cancer suitable for treatment on the basis that the subject has a decreased level and / or activity of regulatory T cells upon administration to the subject of the melanoma cell lysate, and / or an immunotherapeutic extract, component or antigen thereof.

62. A method of treating lung cancer in a human subject, the method including the step of administering to the subject a human vaccinina melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

63. A method of selecting a subject with lung cancer suitable for treatment with a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof, the method including the step of identifying a subject that shows a T cell response upon vaccination of the subject with the melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

64. A method of selecting a subject with lung cancer suitable for treatment with a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof, the method including the step of identifying a subject that has a decreased level and / or activity of regulatory T cells upon vaccination of the subject with a melanoma cell lysate and / or an immunotherapeutic extract, component or antigen thereof.

65. The method of any one of claims 62 to 64, wherein the lung cancer is Stage IV lung cancer.

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