Administration of bacterial compositions for use in the treatment of lung metastases

Abstract

This invention relates to an administration procedure for effectively treating companion animals suffering from lung metastases. In particular, the sprayed BCG composition is delivered directly to the lungs to increase uptake within the lungs and to avoid loss of the active ingredient, for example, in the mucous membrane of the upper respiratory tract.

Description

[Technical Field]

[0001] This invention relates to an administration procedure for effectively treating companion animals suffering from lung metastases. In particular, the sprayed BCG composition is delivered directly to the lungs to increase uptake within the lungs and to avoid loss of the active ingredient, for example, in the mucous membrane of the upper respiratory tract. [Background technology]

[0002] The Bacillus Calmette-Guerin (BCG) vaccine contains live attenuated Mycobacterium bovis bacteria. This vaccine has long been used to prevent tuberculosis (Mycobacterium tuberculosis) in humans. More recently, the BCG vaccine has also been shown to be an effective treatment for several types of tumors in humans.

[0003] The mechanism of action of BCG in cancer treatment is extensive and not fully understood. Components derived from live BCG are well-known agonists / activators of pattern recognition receptors (PRRs) expressed by antigen-presenting cells (dendritic cells, macrophages, and monocytes). When these receptors (Toll-like receptors and NOD receptors) are activated, this induces the downstream production of pro-inflammatory cytokines such as TNF-α, IL-1β, IL-2, IL-6, and IL-12. These cytokines can then stimulate tumor-targeted cytotoxic T cell and NK cell responses, which leads to the removal of cancer cells. In tumor tissue, cancer cells generate an immunological barrier that establishes an immunosuppressive and tumor-tolerant environment. BCG, on the other hand, can break down this barrier by being taken up by macrophages (tumor-associated macrophages), which are abundant in tumor tissue, and shift the immunosuppressive tumor microenvironment toward a pro-inflammatory state.

[0004] In humans, BCG has been shown to induce a positive clinical response in dogs with certain types of cancer. Encouragingly, several routes of administration, including intradermal, subcutaneous, intramuscular, intratumor, and intravenous, have been demonstrated to be safe with low rates of serious side effects. However, lung metastases remain difficult to detect and treat. One challenge with lung metastases is effectively delivering any active ingredient to the diseased site in the lower respiratory tract. Consequently, the prognosis for companion animals such as dogs that develop lung metastases from primary solid tumors is very poor.

[0005] For example, osteosarcoma (OS) is a highly malignant bone tumor in dogs. Despite radical surgical treatment, over 90% of dogs develop lung metastases. Most dogs with OS die as a result of these metastases. As of today, there are no effective treatment options for such metastases once they develop.

[0006] Therefore, it is extremely necessary to provide effective treatment options for lung metastases in companion animals such as dogs.

[0007] In particular, it would be advantageous to provide a treatment that effectively delivers BCG to the patient's lung metastases with minimal systemic exposure and adverse effects. [Overview of the project]

[0008] This invention relates to an improved treatment for lung metastases in companion animals. The treatment includes, for example, the administration of BCG to the patient's lungs by intubation, further adjusted by conditions of positive pressure ventilation. Surprisingly, this treatment has been shown to be efficient in a clinical setting and may provide a solution for patients for whom there are currently no viable treatments.

[0009] Therefore, the object of the present invention relates to the treatment of lung metastases in companion animals.

[0010] Specifically, the object of the present invention is to provide a more efficient method for administering a BCG composition to companion animals suffering from lung metastases.

[0011] Accordingly, one aspect of the present invention relates to a Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, or improvement of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animals.

[0012] Embodiments of the present invention relate to a BCG composition for use as described herein, wherein the BCG composition is administered to the lungs via a tracheal tube.

[0013] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the BCG composition is administered to a companion animal under positive pressure ventilation conditions. [Brief explanation of the drawing]

[0014] [Figure 1] The image shows an X-ray of the lung of canine patient X. A comparison of lung metastases originating from melanoma tumors (white arrows) before the first BCG dose on day 0 (A) and before the fourth BCG dose on day 33 (B) shows a visual reduction in the diameter of the metastatic tissue. [Figure 2] The image shows a CT scan of the lungs of canine patient X. A comparison of lung metastases originating from melanoma tumors (white arrows) before the first BCG dose on day 0 (A) and before the fourth BCG dose on day 33 (B) shows a visual reduction in the diameter of the metastatic tissue. [Figure 3] Figure 2 shows a magnified view of the CT scan. The size of lung metastases was measured using software (A) before the first BCG dose on day 0 and (B) before the fourth BCG dose on day 33. [Figure 4]Plasma secretion of (A) interleukin 8 (IL-8) and (B) interleukin 10 (IL-10) in canine patient X before and after treatment with BCG is shown. IL-8 is a marker of disease burden in melanoma, correlates with prognosis, and the lower the amount of plasma IL-8, the better the prognosis correlates. IL-10 is an anti-inflammatory cytokine, and a decrease in its level indicates reduced immunosuppression.

Mode for Carrying Out the Invention

[0015] The present invention will be described in more detail below.

[0016] Definitions Before summarizing the present invention in more detail, a set of terms and conventions are first defined.

[0017] Primary solid tumor In this context, the term "primary solid tumor" refers to an abnormal mass of tissue that grows uncontrollably. A primary solid tumor is the original or first tumor in the body. A primary solid tumor can spread to other parts of the body and form secondary tumors called metastases. These metastases are the same type of cancer as the primary solid tumor.

[0018] A primary solid tumor can be either benign or malignant. For the treatments disclosed herein, since these tumors are more likely to metastasize than benign tumors, primary solid tumors are typically malignant.

[0019] A primary solid tumor should be distinguished from a liquid tumor that circulates in the body through the bloodstream.

[0020] Lung metastasis In this context, the term "lung metastasis" refers to a secondary tumor located in the lung that has spread from a primary solid tumor. Typically, a lung metastasis is located in the lung parenchyma or pleura.

[0021] The terms "lung metastases" and "pulmonary metastases" may be used interchangeably.

[0022] sarcoma In this context, the term "sarcoma" refers to a malignant tumor of mesenchymal-derived transformed cells (connective tissue). Affected mesenchymal cells can be found in different types of connective tissue, such as bone, cartilage, fat, or vascular tissue. Sarcomas can be divided into two overall groups: osteosarcomas and soft tissue sarcomas.

[0023] Sarcoma is a primary solid tumor that can cause metastasis to other parts of the body, such as the lungs.

[0024] Melanoma In this context, the term "melanoma" refers to cancer that develops in melanin-producing cells called melanocytes. Melanocytes are a type of mesenchymal cell derived from neuronal cells. Therefore, melanoma should be considered a sarcoma.

[0025] cancer In this context, the term "carcinoma" refers to a malignant tumor of epithelial-derived transformed cells. Carcinomas can form in the tissues lining the inner or outer surfaces of the body and include, but are not limited to, breast cancer, prostate cancer, lung cancer, and colorectal cancer.

[0026] Carcinomas can be classified according to their histological type into adenocarcinoma, squamous cell carcinoma, adenosquamous carcinoma, anaplastic carcinoma, large cell carcinoma, and small cell carcinoma.

[0027] In the context of this treatment, carcinomas are considered primary solid tumors that can cause metastasis to other parts of the body, such as the lungs.

[0028] Lymphoma In this context, the term "lymphoma" refers to a primary solid tumor that originates from lymphocytes.

[0029] Bacillus Calmette-Guerin (BCG) composition In this context, the term “BCG composition” refers to any composition containing bioattenuated Mycobacterium bovis BCG as the active ingredient. The BCG composition may contain one or more pharmaceutically acceptable carriers, excipients, and / or diluents to make it suitable for administration to subjects such as non-human patients. The BCG composition may be administered in spray form.

[0030] The BCG composition may be a commercially available BCG vaccine.

[0031] Pharmaceutically acceptable In this context, the term "pharmaceutically acceptable" refers to molecular entities and compositions suitable for use in animals that offer a reasonable benefit-to-risk ratio and do not involve excessive adverse side effects (such as toxicity, irritation, and allergic reactions).

[0032] sprayed and sprayed In this context, the term "aerosol" refers to the process of converting a liquid composition into a fine spray, aerosol, or mist. Therefore, an aerosol composition is in the form of a fine spray, aerosol, or mist.

[0033] Spraying can be achieved using sprayers such as jet sprayers, ultrasonic sprayers, or mesh sprayers, which are connected to a system for mechanical ventilation.

[0034] Mechanical ventilation In this context, the term "mechanical ventilation" refers to the use of ventilation devices to provide artificial ventilation, either fully or partially.

[0035] The terms "mechanical ventilation," "auxiliary ventilation," and "intermittent forced ventilation" may be used interchangeably.

[0036] Companion animals In this context, the term “companion animal” refers to a non-human animal that has been domesticated or raised as a livestock. Companion animals do not include working animals, livestock, or laboratory animals.

[0037] Companion animals include, but are not limited to, dogs, cats, horses, rabbits, hamsters, ferrets, and guinea pigs.

[0038] The terms “companion animal” and “pet” may be used interchangeably.

[0039] about Whenever the term “approximately” is used herein in the context of quantities, such as absolute quantities (e.g., number, purity, concentration, weight, size, etc.) or relative quantities (e.g., percentage, equivalent, or ratio), timeframes, and parameters (e.g., temperature, pressure, etc.), it will be understood that such variables are approximations and, as such, may vary by ±10%, e.g., ±5%, preferably ±2% (e.g., ±1%) from the given actual number. This is true even when such a number is initially presented as a percentage (e.g., “approximately 10%” may mean ±10% of the number 10, which is any of the ranges from 9% to 11%).

[0040] Treatment of lung metastases Unfortunately, the prognosis for companion animals that develop cancer is often poor due to limited treatment options or high treatment costs. The lack of treatment options is even more pronounced for companion animals with metastatic diseases, such as lung metastases, for which no effective treatment is available. Therefore, companion animals with lung metastases are often euthanized immediately after diagnosis or ultimately die from the disease.

[0041] Mycobacterium bovis Bacillus Calmette-Guerin (BCG) is the first live-attenuated vaccine established to prevent childhood meningitis and disseminated tuberculosis (TB). However, BCG has been shown to potentially increase the immune system's ability to fight other pathogens besides TB, for example, by promoting nonspecific responses in both T-cell-mediated adaptive responses and innate immune responses.

[0042] The inventors of this invention have devised a method for efficiently targeting a BCG composition in lung metastases to induce the complete therapeutic effect of Mycobacterium bovis.

[0043] In particular, and surprisingly, it was found that direct administration of the BCG composition to the lungs of companion animal patients significantly reduced tumor burden. Furthermore, it was found that the development of further lung metastases was delayed or prevented.

[0044] Accordingly, one aspect of the present invention relates to a Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, or improvement of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animals.

[0045] It should be understood that BCG administration to the lungs can also be used as a prophylactic treatment to prevent the development of lung metastases. Thus, BCG compositions may be administered at or after the diagnosis of the primary tumor, but may also be administered before the development and / or diagnosis of lung metastases.

[0046] Accordingly, embodiments of the present invention relate to a Bacillus Calmette-Guerin (BCG) composition for use in the prevention of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animal.

[0047] Another embodiment of the present invention relates to a Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, improvement, or prevention of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animal.

[0048] Further embodiments of the present invention relate to BCG compositions for use as described herein, wherein the treatment of lung metastases results in reduction or stabilization of the metastases.

[0049] The active ingredient, intended to exert its effect at a target site in the patient's lungs, is typically delivered in the form of an aerosol or spray composition. The spray composition can be prepared by any conventional means.

[0050] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the BCG composition is a spray BCG composition.

[0051] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the BCG composition is sprayed by pneumatic, mechanical or electrical means.

[0052] However, in non-human animals that cannot be instructed to breathe only through their mouths, delivery to the lungs is complicated. As a result, the therapeutic effect of conventional lung delivery by inhalation may be insufficient because much of the active ingredient is retained in the nasal mucosa. Furthermore, such delivery to non-human animals tends to pose a risk of undesirable exposure of clinicians or animal owners to the active ingredient.

[0053] By directly delivering the BCG composition to the lungs via intubation, it bypasses the upper respiratory tract mucosa (nose, throat, etc.). Therefore, although not constrained by theory, it is assumed that a more efficient distribution of the active ingredient is achieved without dispersing the BCG composition into and around non-infected tissue. Surprisingly, this administration strategy was found to produce a clinically detectable response in canine patients with lung metastases (see Example 1).

[0054] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the BCG composition is administered to the lungs via a tracheal tube.

[0055] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the tracheal tube is an endotracheal tube.

[0056] Further embodiments of the present invention relate to BCG compositions for use as described herein, wherein the route of administration is oral tracheal or nasal tracheal, preferably oral tracheal.

[0057] Mechanical ventilation refers to the use of ventilators to provide artificial ventilation, either fully or partially. Non-invasive mechanical ventilation can be performed on patients who are conscious and able to use a face or nasal mask. Invasive mechanical ventilation refers to situations in which the placement of devices to create an airway, such as an endotracheal tube or nasotracheal tube, is utilized.

[0058] The two main types of mechanical ventilation are positive pressure ventilation, in which air is forced into the lungs through the airways, and negative pressure ventilation, in which air is drawn into the lungs. In this specification, it is assumed that positive pressure ventilation is particularly advantageous for the delivery of BCG compositions to lung metastases.

[0059] Positive pressure ventilation can be performed by increasing the patient's airway pressure through an endotracheal tube. Positive pressure allows air to flow into the airway until the ventilation device stops flowing. The airway pressure then drops to zero, and the elastic recoil of the chest wall and lungs pushes the air out through passive exhalation.

[0060] While not bound by theory, this specification assumes that positive pressure ventilation is preferred because the distal region of the lung is better inflated, and therefore the BCG composition is more completely distributed throughout the lung.

[0061] Accordingly, embodiments of the present invention relate to a BCG composition for use as described herein, wherein the BCG composition is administered to the companion animal under mechanical ventilation conditions.

[0062] A preferred embodiment of the present invention relates to a BCG composition for use as described herein, wherein the BCG composition is administered under positive pressure ventilation conditions.

[0063] While not bound by theory, it is hypothesized that the therapeutic effect of BCG compositions on lung metastases may be due to their ability to efficiently drive the maturation of immune cells such as dendritic cells and macrophages, acting as immunostimulatory components that mobilize them against cancerous tissue. The BCG composition may contain any Mycobacterium bovis strain that induces this maturation; in other words, the BCG composition is not limited to one specific bacterial strain.

[0064] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the BCG composition comprises bioattenuated Mycobacterium bovis BCG.

[0065] Another embodiment of the present invention relates to a BCG composition for use as described in any one of the preceding items, wherein the BCG composition comprises a bacterial strain or substrain selected from the group consisting of Pasteur 1173 P2, Danish 1331, Glaxo 1077, Tokyo 172-1, Russian BCG-I, and Moreau RDJ strain.

[0066] Further embodiments of the present invention relate to BCG compositions for use as described herein, wherein the attenuated Mycobacterium bovis BCG is Danish strain 1331.

[0067] BCG compositions may function with, and are not limited to, any specific, conventional pharmaceutically acceptable carriers, excipients, and diluents. However, several conventional carriers, excipients, and diluents are known to function well specifically with BCG, including, but not limited to, those contained in BCG vaccines. Therefore, BCG compositions may include, or consist of, commercially available BCG vaccines. Such vaccines are known to be safe and free from serious adverse effects.

[0068] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the BCG composition comprises one or more pharmaceutically acceptable carriers, excipients, and / or diluents.

[0069] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein one or more pharmaceutically acceptable carriers, excipients, and / or diluents are selected from the group consisting of sodium glutamate, magnesium sulfate heptahydrate, dipotassium phosphate, citric acid monohydrate, L-asparagine monohydrate, ferric ammonium citrate, and glycerol.

[0070] Other carrier solutions include, but are not limited to, buffered salines such as Hank's equilibrium salt solution, Tris buffered saline, and HEPES buffer solution.

[0071] Yet another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the BCG composition is a BCG vaccine.

[0072] Cancer diseases are widely known to have a poor prognosis, and the degree of the treatment regimen varies among patients. Lung metastases are similar, and it is difficult to predict the required treatment dose. However, the majority of patients require repeated administration of an effective dose to achieve a favorable clinical outcome. The BCG composition herein can be repeatedly administered, if necessary, without any serious adverse effects.

[0073] Therefore, an embodiment of the present invention relates to a BCG composition for use as described herein, wherein the BCG composition is administered as a single effective dose or multiple effective doses.

[0074] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the number of effective doses administered is at least 2, such as at least 3, such as at least 4, such as at least 5, such as at least 6, such as at least 7.

[0075] A further embodiment of the present invention relates to a BCG composition for use as described herein, wherein each effective dose is from about 0.5×10 7 to about 2×10 8 colony forming units (CFU), for example, from about 1×10 7 to about 1.5×10 8 CFU, for example, from about 2×10 7 to about 1×10 8 CFU.

[0076] Yet a further embodiment of the present invention relates to a BCG composition for use as described herein, wherein the concentration of the BCG composition is from about 0.2×10 7 CFU / mL to about 1×10 8 CFU / mL, for example, from about 0.5×10 7 CFU / mL to about 8×107 CFU / mL, preferably about 1 × 10 7 CFU / mL ~ approximately 4 x 10 7 This is within the range of CFU / mL.

[0077] The administration of BCG compositions is relatively rapid and can be reliably carried out once the patient has been properly passivated. Therefore, inconsistent administration and medication caused by anxious or restless companion animals are avoided.

[0078] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the administration of an effective dose to the lungs is completed within a period of about 2 to 15 minutes, for example, about 5 to 10 minutes.

[0079] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the companion animal is sedated during administration of the BCG composition.

[0080] Yet another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the companion animal is anesthetized during administration of the BCG composition.

[0081] Lung metastases are secondary tumors that spread from another primary tumor site in the body to the affected lung. Primary tumors are most commonly solid and can occur in most tissues of the body, including but not limited to bone, breast, lung, prostate, colon, bladder, and kidney. Malignant primary solid tumors, in particular, promote metastasis.

[0082] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein lung metastases originate from a primary solid tumor.

[0083] All primary tumors can metastasize regardless of their origin. Therefore, lung metastases can originate from a wide variety of cancers.

[0084] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the primary solid tumor is selected from the group consisting of sarcomas, carcinomas, lymphomas, and carcinosarcomas.

[0085] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the primary solid tumor is a sarcoma selected from the group consisting of melanoma, soft tissue sarcoma, fibrosarcoma, periangiocarcinoma, peripheral nerve sheet tumor, liposarcoma, myxosarcoma, leiomyosarcoma, rhabdomyosarcoma, synovial cell sarcoma, gastrointestinal stromal tumor, histiocytic sarcoma, angiosarcoma, lymphangiosarcoma, chondrosarcoma, osteosarcoma, and osteochondrosarcoma.

[0086] A preferred embodiment of the present invention relates to a BCG composition for use as described herein, wherein the primary solid tumor is melanoma.

[0087] Further embodiments of the present invention relate to BCG compositions for use as described herein, wherein the primary solid tumors are carcinomas selected from the group consisting of squamous cell carcinoma, nasal adenocarcinoma, thyroid carcinoma and adenocarcinoma, parathyroid carcinoma and adenocarcinoma, gastrointestinal carcinoma and adenocarcinoma, esophageal carcinoma, gastric carcinoma, small intestine carcinoma, colorectal carcinoma, rectal carcinoma, bronchoalveolar carcinoma, lung adenocarcinoma, hepatocellular carcinoma, pancreatic carcinoma and adenocarcinoma, renal cell carcinoma, transitional cell carcinoma, nephroblastoma, prostate adenocarcinoma, uterine carcinoma and adenocarcinoma, Leydig cell tumor, Sertoli cell tumor, seminoma, granulosa cell tumor, teratoma, anal adenocarcinoma and adenocarcinoma, oligodendroglioma, glioblastoma, and meningioma.

[0088] Currently, there are no viable options for treating lung metastases in companion animals, and therefore such diagnoses often lead to euthanasia or premature death. It is assumed that all companion animals would benefit from the treatment scheme described herein. This is because, since not all companion animals have a sufficient degree of consciousness to allow breathing only through the mouth, the active ingredient tends to be retained in the nasal mucosa when administered via a mask as in the conventional method. Therefore, direct administration to the lungs ensures better delivery of the BCG composition to the lung metastatic sites, and even more so when administration is under positive pressure ventilation conditions.

[0089] Companion animals such as dogs, cats, and horses may have relatively longer lifespans than other companion animals, making them more likely to shrink lung metastases and benefit from the treatments described herein.

[0090] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein the companion animal is selected from the group consisting of dogs, cats, horses, rabbits, hamsters, ferrets, and guinea pigs.

[0091] A preferred embodiment of the present invention relates to a BCG composition for use as described herein, wherein the companion animal is a dog.

[0092] Another embodiment of the present invention relates to a BCG composition for use as described herein, wherein the companion animal is a cat.

[0093] Further embodiments of the present invention relate to a BCG composition for use as described herein, wherein the companion animal is a horse.

[0094] The treatments described herein have been shown to reduce the tumor burden of lung metastases. The tumor burden may be assessed based on several parameters, such as the size of lung metastases and / or the levels of known prognostic immune indicators or biomarkers.

[0095] Known tumor burden biomarkers include, but are not limited to, interleukin-8 (IL-8) and interleukin-10 (IL-10). IL-8 is a known biomarker of tumor burden, and high levels of IL-8 are associated with high tumor burden. IL-8 is produced by several cancer types, including melanoma, and is involved in pro-neoplastic angiogenesis and inflammation. IL-10 is an anti-inflammatory cytokine. High levels of IL-10 exhibit tumor-induced immunosuppression.

[0096] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, wherein administration of the BCG composition results in a reduction of tumor burden.

[0097] Another embodiment of the present invention relates to the BCG composition for use described in item X29, wherein the reduction of tumor burden is one or more selected from the group consisting of a reduction in the size of lung metastases, a slowing of metastatic growth rate, and a reduction in the levels of interleukin-8 (IL-8) and / or interleukin-10 (IL-10) concentrations in the serum of the companion animal.

[0098] As with many other cancer treatments, this administration of the BCG composition may be combined with the administration of another immunotherapy composition. The immunotherapy composition may be a commercially available composition or product.

[0099] Accordingly, embodiments of the present invention relate to BCG compositions for use as described herein, and further include the administration of immunotherapeutic compositions.

[0100] Another aspect of the present invention relates to a method for treating, inhibiting, or improving lung metastases in companion animals, the method comprising administering a BCG composition to the lungs of the companion animals.

[0101] The BCG composition may be conveniently provided as a kit ready to be loaded into an intubation system used to deliver the BCG composition to the lungs of a companion animal. The intubation system comprises all the components necessary to deliver the BCG composition to the lungs, including, but not limited to, a spray chamber for loading the BCG composition into the associated tube for patient intubation, and a circuit for maintaining the patient in anesthesia during administration of the BCG composition.

[0102] Therefore, one aspect of the present invention is: Bacillus Calmette-Guerin (BCG) composition and An intubation system suitable for use with companion animals, Regarding the kit, including.

[0103] Another aspect of the present invention relates to an intubation system suitable for use with companion animals, wherein the intubation system is loaded with a Bacillus Calmette-Guerin (BCG) composition.

[0104] Any list or discussion of documents that have clearly been previously published in this specification should not necessarily be considered an endorsement that such documents are part of the latest technology or common knowledge.

[0105] Any preferences, choices, and embodiments relating to a given aspect, feature, or parameter of the present invention should be considered to be disclosed in conjunction with all other preferences, choices, and embodiments relating to all other aspects, features, and parameters of the present invention, unless otherwise indicated by the context. This is particularly true of descriptions of BCG compositions, tracheal tubes, and mechanical ventilation described in relation to medical uses or therapeutic methods utilizing BCG compositions, and such embodiments may readily be part of the kits and intubation systems described herein. Embodiments and features of the present invention are also outlined in the following sections.

[0106] item X1. A Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, improvement, or prevention of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animal.

[0107] X1a. A Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, or improvement of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of the companion animals.

[0108] X2. The BCG composition for use according to either item X1 or X1a, wherein the BCG composition is administered to the lungs via a tracheal tube.

[0109] X3. The BCG composition for use according to any one of items X1, X1a, or X2, wherein the tracheal tube is an endotracheal tube.

[0110] X4. A BCG composition for use according to any one of the above items, wherein the route of administration is oral tracheal or nasal tracheal, preferably oral tracheal.

[0111] X5. The BCG composition for use according to any one of the above items, wherein the BCG composition is administered to the companion animal under conditions of mechanical ventilation.

[0112] X6. The BCG composition according to any one of the above items, wherein the BCG composition is administered under conditions of positive pressure ventilation.

[0113] X7. A BCG composition for use according to any one of the above items, wherein the BCG composition is a spray BCG composition.

[0114] X8. The BCG composition for use according to item X7, wherein the BCG composition is sprayed by pneumatic, mechanical, or electrical means.

[0115] X9. A BCG composition for use according to any one of the above items, wherein the BCG composition comprises bioattenuated Mycobacterium bovis BCG.

[0116] X10. A BCG composition for use according to any one of the above items, wherein the BCG composition comprises a bacterial strain or substrain selected from the group consisting of Pasteur 1173 P2, Danish 1331, Glaxo 1077, Tokyo 172-1, Russian BCG-I, and Moreau RDJ strain.

[0117] X11. A BCG composition for use according to either item X9 or X10, wherein the attenuated Mycobacterium bovis BCG is Danish strain 1331.

[0118] X12. The BCG composition for use according to any one of the above items, wherein the BCG composition comprises one or more pharmaceutically acceptable carriers, excipients, and / or diluents.

[0119] X13. The BCG composition for use according to item X12, wherein the one or more pharmaceutically acceptable carriers, excipients, and / or diluents are selected from the group consisting of sodium glutamate, magnesium sulfate heptahydrate, dipotassium phosphate, citric acid monohydrate, L-asparagine monohydrate, ferric ammonium citrate, and glycerol.

[0120] X14. A BCG composition for use according to any one of the above items, wherein the BCG composition is a BCG vaccine.

[0121] X15. The BCG composition for use according to any one of the above items, wherein the BCG composition is administered as a single effective dose or as multiple effective doses.

[0122] X16. The BCG composition for use according to item X15, wherein the number of effective doses administered is at least 2, e.g., at least 3, e.g., at least 4, e.g., at least 5, e.g., at least 6, e.g., at least 7.

[0123] X17. Each effective dose is approximately 0.5 × 10 7 ~Approx. 2×10 8 Colony-forming units (CFUs), for example, approximately 1 × 10⁻⁶ 7 ~Approx. 1.5×10 8 CFU, for example, approximately 2 × 10 7 ~Approx. 1×10 8 A BCG composition for use as described in either item X15 or X16, which is within the range of CFU.

[0124] X18. A BCG composition for use according to any one of items X15 to X17, wherein the administration of the effective dose to the lungs is completed within a period of about 2 to about 15 minutes, for example, about 5 to about 10 minutes.

[0125] X19. The BCG composition for use according to any one of the items, wherein the companion animal is sedated during the administration of the BCG composition.

[0126] X20. The BCG composition for use according to any one of the items, wherein the companion animal is anesthetized during the administration of the BCG composition.

[0127] X21. The BCG composition for use according to any one of the above items, wherein the lung metastases originate from a primary solid tumor.

[0128] X22. The BCG composition for use according to item X21, wherein the primary solid tumor is selected from the group consisting of sarcoma, carcinoma, lymphoma, and carcinosarcoma.

[0129] X23. A BCG composition for use according to either item X21 or X22, wherein the primary solid tumor is a sarcoma selected from the group consisting of melanoma, soft tissue sarcoma, fibrosarcoma, periangiocarcinoma, peripheral nerve sheet tumor, liposarcoma, myxosarcoma, leiomyosarcoma, rhabdomyosarcoma, synovial cell sarcoma, gastrointestinal stromal tumor, histiocytic sarcoma, angiosarcoma, lymphangiosarcoma, chondrosarcoma, osteosarcoma, and osteochondrosarcoma.

[0130] X24. The BCG composition for use according to any one of items X21 to X23, wherein the primary solid tumor is melanoma.

[0131] X25. A BCG composition for use as described in either item X21 or X22, wherein the primary solid tumor is a carcinoma selected from the group consisting of squamous cell carcinoma, nasal adenocarcinoma, thyroid carcinoma and adenocarcinoma, parathyroid carcinoma and adenocarcinoma, gastrointestinal carcinoma and adenocarcinoma, esophageal carcinoma, gastric carcinoma, small intestine carcinoma, colorectal carcinoma, rectal carcinoma, bronchoalveolar carcinoma, lung adenocarcinoma, hepatocellular carcinoma, pancreatic carcinoma and adenocarcinoma, renal cell carcinoma, transitional cell carcinoma, nephroblastoma, prostate adenocarcinoma, uterine carcinoma and adenocarcinoma, Leydig cell tumor, Sertoli cell tumor, seminoma, granulosa cell tumor, teratoma, anal adenocarcinoma and adenocarcinoma, oligodendroglioma, glioblastoma, and meningioma.

[0132] X26. A BCG composition for use according to any one of the above items, wherein the companion animal is selected from the group consisting of dogs, cats, horses, rabbits, hamsters, ferrets, and guinea pigs.

[0133] X27. The BCG composition for use according to any one of the preceding items, wherein the companion animal is a dog.

[0134] X28. The BCG composition for use according to any one of items X1 to X26, wherein the companion animal is a cat.

[0135] X29. A BCG composition for use according to any one of the above items, wherein the administration of the BCG composition results in a reduction of tumor burden.

[0136] X30. The BCG composition for use as described in item X29, wherein the reduction in tumor burden is one or more selected from the group consisting of a reduction in the size of lung metastases, a slowing of metastatic growth rate, and a reduction in the levels of interleukin-8 (IL-8) and / or interleukin-10 (IL-10) concentrations in the serum of the companion animal.

[0137] X31. A BCG composition for use according to any one of the preceding items, further comprising the administration of an immunotherapy composition.

[0138] Y1. A method for treating, inhibiting, improving, or preventing lung metastases in a companion animal, comprising administering a BCG composition to the lungs of the companion animal.

[0139] Y1a. A method for treating, inhibiting, or improving lung metastases in a companion animal, comprising administering a BCG composition to the lungs of the companion animal.

[0140] Y2. The method according to either item Y1 or Y1a, wherein the BCG composition is administered to the lungs via a tracheal tube.

[0141] Y3. The method according to any one of items Y1, Y1a, or Y2, wherein the tracheal tube is an endotracheal tube.

[0142] Y4. The method according to any one of items Y1 to Y3, wherein the route of administration is oral tracheal or nasal tracheal, preferably oral tracheal.

[0143] Y5. The method according to any one of items Y1 to Y4, wherein the BCG composition is administered to the companion animal under mechanical ventilation conditions.

[0144] Y6. The method according to any one of items Y1 to Y5, wherein the BCG composition is administered under positive pressure ventilation conditions.

[0145] Y7. The method according to any one of items Y1 to Y6, wherein the BCG composition is a spray BCG composition.

[0146] Y8. The method according to item Y7, wherein the BCG composition is sprayed by air pressure, mechanical or electrical means.

[0147] Y9. The method according to any one of items Y1 to Y8, wherein the BCG composition comprises bio-attenuated Mycobacterium bovis BCG.

[0148] Y10. The method according to any one of items Y1 to Y9, wherein the BCG composition comprises a bacterial strain or substrain selected from the group consisting of Pasteur 1173 P2, Danish 1331, Glaxo 1077, Tokyo 172-1, Russian BCG-I, and Moreau RDJ strain.

[0149] Y11. The method according to either item Y9 or Y10, wherein the attenuated Mycobacterium bovis BCG is Danish strain 1331.

[0150] Y12. The method according to any one of items Y1 to Y11, wherein the BCG composition comprises one or more pharmaceutically acceptable carriers, excipients, and / or diluents.

[0151] Y13. The method according to item Y12, wherein the one or more pharmaceutically acceptable carriers, excipients, and / or diluents are selected from the group consisting of sodium glutamate, magnesium sulfate heptahydrate, dipotassium phosphate, citric acid monohydrate, L-asparagine monohydrate, ferric ammonium citrate, and glycerol.

[0152] Y14. The method according to any one of items Y1 to Y13, wherein the BCG composition is a BCG vaccine.

[0153] Y15. The method according to any one of items Y1 to Y14, wherein the BCG composition is administered as a single effective dose or as multiple effective doses.

[0154] Y16. The method according to item Y15, wherein the number of effective doses administered is at least 2, e.g., at least 3, e.g., at least 4, e.g., at least 5, e.g., at least 6, e.g., at least 7.

[0155] Y17. Each effective dose is approximately 0.5 × 10 7 ~Approx. 2×10 8 Colony-forming units (CFUs), for example, approximately 1 × 10⁻⁶ 7 ~Approx. 1.5×10 8 CFU, for example, approximately 2 × 10 7 ~Approx. 1×10 8 The method described in either item Y15 or Y16, which is the range of CFU.

[0156] Y18. The method according to any one of items Y15 to Y17, wherein the administration of the effective dose to the lungs is completed within a period of approximately 2 to 15 minutes, for example, approximately 5 to 10 minutes.

[0157] Y19. The method according to any one of items Y1 to Y18, wherein the companion animal is sedated during the administration of the BCG composition.

[0158] Y20. The method according to any one of items Y1 to Y19, wherein the companion animal is anesthetized during the administration of the BCG composition.

[0159] Y21. The method according to any one of items Y1 to Y20, wherein the lung metastasis originates from a primary solid tumor.

[0160] Y22. The method according to item Y21, wherein the primary solid tumor is selected from the group consisting of sarcoma, carcinoma, lymphoma, and carcinosarcoma.

[0161] Y23. The method according to either item Y21 or Y22, wherein the primary solid tumor is a sarcoma selected from the group consisting of melanoma, soft tissue sarcoma, fibrosarcoma, periangiocarcinoma, peripheral nerve sheet tumor, liposarcoma, myxosarcoma, leiomyosarcoma, rhabdomyosarcoma, synovial cell sarcoma, gastrointestinal stromal tumor, histiocytic sarcoma, angiosarcoma, lymphangiosarcoma, chondrosarcoma, osteosarcoma, and osteochondrosarcoma.

[0162] Y24. The method according to any one of items Y21 to Y23, wherein the primary solid tumor is melanoma.

[0163] Y25. The method according to either item Y21 or Y22, wherein the primary solid tumor is a carcinoma selected from the group consisting of squamous cell carcinoma, nasal adenocarcinoma, thyroid carcinoma and adenocarcinoma, parathyroid carcinoma and adenocarcinoma, gastrointestinal carcinoma and adenocarcinoma, esophageal carcinoma, gastric carcinoma, small intestine carcinoma, colorectal carcinoma, rectal carcinoma, bronchoalveolar carcinoma, lung adenocarcinoma, hepatocellular carcinoma, pancreatic carcinoma and adenocarcinoma, renal cell carcinoma, transitional cell carcinoma, nephroblastoma, prostate adenocarcinoma, uterine carcinoma and adenocarcinoma, Leydig cell tumor, Sertoli cell tumor, seminoma, granulosa cell tumor, teratoma, anal adenocarcinoma and adenocarcinoma, oligodendroglioma, glioblastoma, and meningioma.

[0164] Y26. The method according to any one of items Y1 to Y25, wherein the companion animal is selected from the group consisting of dogs, cats, horses, rabbits, hamsters, ferrets, and guinea pigs.

[0165] Y27. The method according to any one of items Y1 to Y26, wherein the companion animal is a dog.

[0166] Y28. The method according to any one of items Y1 to Y26, wherein the companion animal is a cat.

[0167] Y29. The method according to any one of items Y1 to Y28, wherein administration of the BCG composition results in a reduction of tumor burden.

[0168] Y30. The method according to item Y29, wherein the reduction in tumor burden is one or more selected from the group consisting of a reduction in the size of lung metastases, a slowing of metastatic growth rate, and a reduction in the levels of interleukin-8 (IL-8) and / or interleukin-10 (IL-10) concentrations in the serum of the companion animal.

[0169] Y31. The method according to any one of items Y1 to Y30, further comprising the administration of an immunotherapy composition.

[0170] Z1. Kit, Bacillus Calmette-Guerin (BCG) composition and An intubation system suitable for use with companion animals, A kit that includes this.

[0171] Z2. The intubation system comprising a spray chamber and a tracheal tube, as described in item Z1.

[0172] Z3. The intubation system comprising a spray chamber, a tracheal tube, and an anesthesia circuit, as described in item Z1 or Z2.

[0173] W1. An intubation system suitable for use with companion animals, wherein the intubation system is loaded with a Bacillus Calmette-Guerin (BCG) composition.

[0174] W2. The intubation system according to item W1, comprising a spray chamber and a tracheal tube.

[0175] W3. The intubation system according to either item W1 or W2, wherein the intubation system comprises a spray chamber, a tracheal tube, and an anesthesia circuit.

[0176] W4. The intubation system according to any one of items W1 to W3, wherein the spray chamber is loaded with the BCG composition. [Examples]

[0177] Example 1: Treatment of lung metastases in a canine patient This example describes the treatment and evaluation of clinical outcomes in a canine patient with lung metastases ("Canine Patient X"). In particular, spray BCG was administered via intubation of the patient.

[0178] method patient Canine patient X was a mixed breed dog weighing approximately 8 kg with lung metastases from a solid primary melanoma tumor. The tumor originated in the oral cavity (oral malignant melanoma). The primary tumor was surgically removed (partial mandibular resection), and the draining lymph nodes were removed. Postoperatively, canine patient X developed lung metastases.

[0179] BCG spraying: BCG solution (Danish strain 1331) was diluted in phosphate-buffered saline (PBS) to the desired concentration calculated according to the canine patient's body weight, and approximately 4 mL was used in the procedure (see Table 1 below). The sprayed BCG dose was 2.5 × 10⁶ for the first two doses. 7 From CFU, 3.5 × 10 for the 3rd and 4th doses. 7 The dosage was increased to CFU. For dogs weighing over 20 kg, a dual-concentration dose is appropriate.

[0180] Next, the reconstituted BCG (BCG nebulizer solution) was drawn into a single 5 mL syringe and transferred to the nebulizer chamber of a nebulizer system (Aerogen® Solo, Hamilton Medical). The nebulizer system comprises a nebulizer, a T-piece, an optional breathing circuit adapter, and a connecting cable. [Table 1] Table 1: BCG dose calculation.

[0181] Administration of BCG to canine patients A non-rebreathing T-piece system was used as the anesthesia circuit to deliver aerosolized BCG. The fresh gas flow rate was calculated based on the size of canine patient X (approximately 200 mL / kg / min).

[0182] Before anesthesia was induced by intravenous administration of propofol (1-4 mg / kg) over 2 minutes, the canine patient was sedated with butorphanol (0.2 mg / kg intramuscularly) and given 100% oxygen through a mask. Once autonomic reflexes had ceased, the larynx was visualized using a laryngoscope and an endotracheal tube placed in the trachea. The endotracheal tube cuff was manually inflated with a syringe to ensure there were no air leaks. The canine patient was then connected to a ventilator, which was then turned on to initiate positive pressure ventilation. Total intravenous anesthesia (TIVA) (propofol) was administered to maintain a mild to moderate level of anesthesia. The patient was ventilated using positive pressure ventilation throughout the procedure.

[0183] The canine patients were monitored throughout the procedure using capnography, pulse oximetry, electrocardiogram, blood pressure, and body temperature. The ventilation system was set to ventilate the canine patients with a tidal volume of approximately 10 mL / kg, at a rate of 10–15 breaths / min, with a maximum inspiratory pressure of 15 cm H2O (+ / - 5 cm H2O) and a maximum expiratory pressure of 0 cm H2O (+ / - 5 cm H2O).

[0184] Prior to initiating BCG administration, the canine patients received a single dose of diphenhydramine (1 mg / kg intravenously) and acetaminophen (10 mg / kg intravenously).

[0185] After transferring the BCG spray solution to the spray chamber, the spray unit was switched on. The operator confirmed that the BCG solution was being sprayed by visually inspecting the mist filling the ventilation tube. BCG administration continued until the spray unit was empty (approximately 4-8 minutes).

[0186] Once the spray chamber was empty, the sprayer was turned off, and the dog patient was removed from the ventilator for 2-3 minutes until it began breathing spontaneously, allowing any remaining aerosol in the circuit to be removed. The dog patient was then disconnected from the anesthesia circuit, and propofol was discontinued.

[0187] Once the canine patient began to regain its autonomic reflexes, the endotracheal tube was removed, and the canine patient was monitored until it regained normal function before being returned to its kennel. During this time, the canine patient was continuously monitored.

[0188] X-ray imaging Canine patient X was imaged while awake. The thoracic cavity was imaged from four different viewpoints: right lateral recumbent, left lateral recumbent, sternal lateral recumbent, and dorsal lateral recumbent. Imaging settings were adjusted according to the patient's weight. Images were evaluated by a committee-certified veterinary radiologist. The size of lung metastases was subjectively assessed and compared between time points using image analysis software (Vue PACS v12, Philips).

[0189] CT scan Canine patient X underwent computed tomography (CT) scanning using a CT scanner (Revolution® CT scanner, GE HealthCare) while under anesthesia prior to BCG spraying. The patient was placed in the lateral sternal position. Images were acquired for both soft tissue and bone alignment before and after administration of intravenous contrast agent solution (Iohexol). Imaging settings were adjusted according to the patient's weight. Images were evaluated by a committee-certified veterinary radiologist. Lung metastasis size was assessed between time points using image analysis software (Vue PACS v12, Philips).

[0190] Cytokine analysis Serum levels of IL-8 and IL-10 in canine patient X were determined using multiplex analysis (ProcartaPlex® Canine Cytokine Chemokine Growth Factor Panel 1 11-Plex, Invitrogen). Multiplex analysis was used for the following purposes: 1. Short-term blood sampling (2-96 hours after BCG spray treatment). Analysis of the effect of BCG spray treatment at the time of administration, with the aim of detecting correlations with potential side effects that may occur during administration.

[0191] 2. Short-term blood sampling (2-96 hours after BCG spray treatment). Identification of any possible correlations between local BCG administration and changes in the pulmonary and systemic circulating cytokine profiles.

[0192] 3. Long-term blood sampling (at baseline, every two weeks until the end of treatment, and then at follow-up visits). Identification of any possible correlations between changes in cytokine levels and reductions in tumor burden (e.g., reduction in the number and size of metastases).

[0193] In short, blood samples were obtained from dog patient X, and 0.5 mL of blood collected in an EDTA-treated tube was separated by centrifugation at 1,000 × g for 10 minutes at room temperature. The top layer corresponding to the plasma fraction was then transferred to a new tube and immediately stored at -80°C until multiplex analysis.

[0194] For the multiplex assay, frozen plasma samples were thawed, mixed by vortexing, and pelletized by centrifugation at 10,000 × g for 10 minutes. The clear supernatant was then transferred to a new tube, and 25 μL of each sample, along with the standard and blank, was double-dispensed onto a 96-well plate pre-loaded with capture beads, according to the manufacturer's instructions. The plate was incubated at room temperature for 2 hours to allow the analytes present in the standard and sample to bind to their respective beads. The plate was then washed, 25 μL of detection antibody mixture was added to each well, and the plate was incubated at room temperature with shaking for 30 minutes. The plate was then washed, 50 μL of streptavidin-PE was added to each well, and the plate was incubated at room temperature with shaking for 30 minutes. The plate was then washed, 120 μL of Reading buffer was added, and the plate was shaken at room temperature for 5 minutes. Next, the plate was inserted into a MAGPIX Luminex instrument (Luminex, DiaSorin), data was acquired, and analyzed using xPONENT software (Luminex, DiaSorin). Subsequently, a graphical presentation of the results was generated using GraphPad Prism software.

[0195] result Canine patient X was enrolled for treatment of lung metastases caused by melanoma. To provide a baseline of disease burden, X-ray and CT scans were taken before initiating treatment (Figures 1A and 2A). What is evident from the X-ray and CT scans on day 0 is the metastasis in the lungs of canine patient X (white arrows).

[0196] Dog patient X had 2.5 × 10 7 CFU, 2.5 x 10 7 CFU, 3.5 x 10 7 CFU, and 3.5 × 10 7 BCG was administered at CFU dose concentrations on days 0, 9, 22, and 33 of each administration cycle. The administration was carried out as described above.

[0197] On day 33 (before the fourth dose), X-ray and CT scans were taken to assess the post-treatment lung metastasis status (Figures 1B and 2B). The X-ray images show a visual reduction in the size of the largest metastasis in the cranial chest between day 0 and day 33. Similarly, the CT scan shows a measurable reduction in the size of metastases in the right cranial lobe (Figure 3).

[0198] A promising reduction in metastasis size was supported by measurements of interleukin-8 (IL-8) and interleukin-10 (IL-10) in the serum of canine patient X.

[0199] Measurements showed a substantial decrease in serum levels of both IL-8 (Figure 4A) and IL-10 (Figure 4B) in canine patient X after the initiation of BCG treatment. Therefore, the cytokine profile suggests less immunosuppression in canine patient X with reduced tumor burden after BCG treatment. Furthermore, the matched cytokine profile did not show any adverse effects associated with direct administration of BCG into the lungs of canine patient X. Therefore, the cytokine data demonstrate a positive effect of the treatment.

[0200] Canine patient X was euthanized 183 days after being diagnosed with metastasis. This significantly exceeds the 54-day median expected survival time (MST) for stage IV melanoma. Canine patient X progressed very well during treatment, and the severity of adverse effects recorded during treatment was lower compared to dogs treated with standard chemotherapy. Adverse effects included a mild increase in body temperature (though not defined as fever) on the first day after treatment, and a mild increase in respiratory rate the following day in some cases. During the final month before euthanasia, canine patient X showed some decrease in appetite and activity levels. Ultimately, canine patient X was euthanized due to seizures caused by brain metastases.

[0201] conclusion Treatment of canine patient X provided a clear, clinically measurable response to the treatment, as demonstrated by a reduction in metastatic size and tumor burden. This canine patient is significantly ahead of the expected median survival time.

[0202] Example 2: Multiplex analysis of cytokine response in dogs treated with BCG administered by different protocols. The objective of this embodiment is to evaluate the level of serum inflammatory markers (such as IL-12) as a function of the BCG delivery method.

[0203] method BCG administration Healthy dogs are treated with indirect and direct delivery methods of spray BCG. The first treatment scheme involves treatment using the direct method first (intubation and positive pressure ventilation under anesthesia), followed by the indirect method 72 hours later (spontaneous ventilation while conscious). The second treatment scheme involves treatment using the indirect method first, followed by the direct method 72 hours later. The same dose of BCG is administered for each delivery method.

[0204] The direct delivery method of sprayed BCG is the same as that described in Example 1. The indirect delivery method of sprayed BCG is performed in conscious dogs using an AeroDog chamber placed over the dog's nasal cavity and connected to an Aerogen Solo® T-piece. The sprayer is operated until the chamber is empty.

[0205] Cytokine analysis Serum levels of inflammatory cytokines in canine patients are determined using multiplex analysis (ProcartaPlex® Canine Cytokine Chemokine Growth Factor Panel 1 11-Plex, Invitrogen) as described in Example 1. Blood samples are collected at baseline (before spraying) and between 2 and 24 hours after BCG spraying, stored at -80°C, and processed for multiplex analysis as described in Example 1.

[0206] The levels of inflammatory cytokines such as IL-12 are determined and compared across different treatment schemes.

[0207] result Levels of several inflammatory cytokines, such as IL-12, are expected to reach higher levels 12 hours after BCG spraying using direct delivery methods compared to indirect methods.

[0208] conclusion Direct delivery of sprayed BCG induces a more pronounced inflammatory response, suggesting a stronger antitumor response.

[0209] Example 3: Treatment of lung metastases in a second canine patient This example describes the treatment and evaluation of clinical outcomes in a canine patient with lung metastases ("Canine Patient Y"). In particular, spray BCG was administered via intubation of the patient.

[0210] method patient Canine patient Y was a 3.5-year-old female Golden Retriever who underwent limb amputation due to chondroblastic osteosarcoma of the distal radius. The diagnosis of osteosarcoma was made one month prior to the amputation. Following the amputation, canine patient Y received multiple rounds of chemotherapy with carboplatin.

[0211] The prognosis for osteosarcoma without treatment is very poor, and even with amputation, most canine patients develop large lung metastases within a few months of amputation. Typically, such patients are euthanized within six months of diagnosis if not treated with chemotherapy, while with chemotherapy, metastasis usually occurs, and survival before the dog shows signs of metastasis is extended to 8-12 months. However, once metastasis develops, the prognosis is poor, and the median expected survival time (MST) varies from 72 to 89 days depending on treatment.

[0212] treatment Canine patient Y received the same treatment as canine patient X (see Example 1). Canine patient Y received a total of three BCG sprays.

[0213] result Canine patient Y was enrolled for treatment of lung metastases caused by osteosarcoma and received initial BCG spray therapy five months after the diagnosis of osteosarcoma. At the time of the initial treatment, two distinct lung metastases were visible on a CT scan.

[0214] 71 days after the initial treatment, a CT scan was performed on dog patient Y after it had received three treatments to assess the status of lung metastases. The CT scan revealed no new metastases.

[0215] Unfortunately, dog patient Y was unable to tolerate the anesthesia well, and it was agreed with the patient's owner that treatment should be discontinued.

[0216] On day 101 of the initial treatment and diagnosis of two lung metastases, canine patient Y was euthanized due to poor gait and reduced activity level. A postmortem chest CT scan showed no new metastases. Canine patient Y's clinical symptoms were not considered to be related to the two lung metastases.

[0217] conclusion Data from canine patient Y indicate that BCG spray treatment protects against the development of new metastases. Canine patient Y survived substantially longer than the prognosis for the indication (i.e., MST was 72–89 days), and the decision to euthanize the patient was not due to clinical symptoms caused by lung metastases.

Claims

1. A Bacillus Calmette-Guerin (BCG) composition for use in the treatment, inhibition, improvement, or prevention of lung metastases in companion animals, wherein the BCG composition is administered to the lungs of a companion animal such as a dog or a cat.

2. The BCG composition for use according to claim 1, wherein the BCG composition is administered to the lungs via a tracheal tube.

3. The BCG composition for use according to claim 1 or 2, wherein the route of administration is oral tracheal or nasal tracheal, preferably oral tracheal.

4. The BCG composition for use according to any one of claims 1 to 3, wherein the BCG composition is administered to the companion animal under mechanical ventilation conditions.

5. The BCG composition for use according to any one of claims 1 to 4, wherein the BCG composition is administered under positive pressure ventilation conditions.

6. The BCG composition for use according to any one of claims 1 to 5, wherein the BCG composition is a spray BCG composition.

7. The BCG composition for use according to any one of claims 1 to 6, wherein the BCG composition comprises bio-attenuated Mycobacterium bovis BCG.

8. The BCG composition for use according to any one of claims 1 to 7, wherein the BCG composition is administered as a single effective dose or as multiple effective doses.

9. The BCG composition for use according to claim 8, wherein the administration of the effective dose to the lungs is completed within a period of about 2 to 15 minutes, for example, about 5 to 10 minutes.

10. The BCG composition for use according to any one of claims 1 to 9, wherein the lung metastases originate from a primary solid tumor.

11. The BCG composition for use according to claim 10, wherein the primary solid tumor is selected from the group consisting of sarcoma, carcinoma, lymphoma, and carcinosarcoma.

12. The BCG composition for use according to any one of claims 1 to 11, wherein the companion animal is selected from the group consisting of dogs, cats, horses, rabbits, hamsters, ferrets, and guinea pigs.

13. The BCG composition for use according to any one of claims 1 to 12, wherein the companion animal is a dog or a cat, preferably a dog.

14. The BCG composition for use according to any one of claims 1 to 13, wherein administration of the BCG composition results in a reduction of tumor burden.

15. The BCG composition for use according to claim 14, wherein the reduction in tumor burden is one or more selected from the group consisting of a reduction in the size of lung metastases, a slowing of metastatic growth rate, and a reduction in the levels of interleukin-8 (IL-8) and / or interleukin-10 (IL-10) concentrations in the serum of the companion animal.

16. It's a kit, Bacillus Calmette-Guerin (BCG) composition and An intubation system suitable for use with companion animals, A kit that includes this.

17. The kit according to claim 16, wherein the intubation system comprises a spray chamber and a tracheal tube.

18. An intubation system suitable for use with companion animals, wherein the intubation system is loaded with a Bacillus Calmette-Guerin (BCG) composition.

19. The intubation system according to claim 18, wherein the intubation system comprises a spray chamber and a tracheal tube.

20. The intubation system according to claim 18 or 19, wherein the BCG composition is loaded into the spray chamber.

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