Fecal material for the treatment of cachexia

Abstract

The present invention relates to the use of allogeneic fecal material for the treatment of cachexia and / or cancer, the allogeneic fecal material being obtained from at least one donor subject, the at least one donor subject having a blood glucose level of at least 30 kg / m 2 a body mass index (BMI) of less than 100 mg / dL, a HOMA-IR value of less than 2.5 mg / dL, and / or an age of less than 60 years. Fecal material from said at least one donor subject may be used to treat cancer and / or related conditions.

Description

[Technical Field]

[0001] The present invention relates to the use of fecal material for the treatment of cachexia, in particular cancer-associated cachexia. [Background technology]

[0002] Cancer is a group of diseases in which somatic cells grow abnormally, resulting in the formation of tumors. Unlike benign tumors, tumors caused by cancer are malignant and have the potential to metastasize and / or spread to other parts of the body. Cancer is a type of neoplasia in which cell growth is uncoordinated with surrounding healthy tissue. Cancer cells are capable of unlimited proliferation and evade programmed cell death. Survival rates depend largely on the type of cancer, the time of diagnosis, and the availability of curative treatments.

[0003] Patients diagnosed with cancer often receive various types of curative treatment, which can be divided into local treatments, including surgery and radiation therapy, and systemic treatments, including immunotherapy, targeted therapy, and chemotherapy. Another type of treatment for patients diagnosed with cancer is palliative care. This type of treatment aims to reduce the cancer patient's suffering and improve their quality of life. Palliative care for cancer patients is most often, although not exclusively, associated with terminal cancer, where the cancer cannot be cured and the patient is expected to die. Palliative care can be combined with curative treatment.

[0004] Curative and palliative cancer treatment can be complicated by cachexia, a complex syndrome resulting in muscle loss. Cachexia, also known as wasting syndrome, can result from a variety of medical conditions but is most commonly associated with end-stage cancer and is known as cancer cachexia. Unlike sarcopenia, a condition characterized by a loss of skeletal muscle mass and function, cachexia cannot be treated with precise and adequate nutritional administration. Cachexia is particularly prevalent in patients diagnosed with gastrointestinal and pancreatic cancers. Cachexia affects four out of five terminally or terminally ill cancer patients. Cachexia in cancer patients reduces quality of life and increases mortality.

[0005] Herremans et al. (2019, Int J Mol Sci) disclose that an imbalance in the gut microbiota, known as dysbiosis, has been shown to influence cancer cachexia. The cancer cachexia phenotype is associated with decreased levels of Lactobacillaceae and increased levels of Enterobacteriaceae and Parabacteroides.

[0006] WO 2016 / 196605 discloses methods for treating and / or preventing cancer through manipulation of commensal microbiota (also known as microflora). In particular, the amount, identity, presence, and / or proportions of microbiota (e.g., gut microbiota) in a subject are manipulated to facilitate one or more concurrent therapies.

[0007] WO 2014 / 121298 discloses administering to a subject a therapeutic composition comprising a purified population of spore-forming bacteria produced by providing fecal material and subjecting the material to a treatment process resulting in the purification of the spore-forming bacteria.

[0008] WO 2019 / 171012 discloses stool collection and sample preparation methods for fecal microbiota transplantation. The resulting homogenous mixture of fecal microbiota can be used for the treatment of intestinal dysbiosis and pathologies associated with such dysbiosis. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] International Publication No. 2016 / 196605 [Patent Document 2] International Publication No. 2014 / 1212982 [Patent Document 3] International Publication No. 2019 / 171012 [Non-patent literature]

[0010] [Non-Patent Document 1] Herremans et al., (2019, Int J Mol Sci) [Non-patent document 2] Biagi et al. (PLoS One. 2010 May 17;5(5):e10667) [Non-patent document 3] Applied Anatomy and Biomechanics in Sport, by T.R. Ackland, B. Elliott, and J. Bloomfield, 2nd ed., 2009 [Non-patent document 4] Clarke et al., Exercise and associated dietary extremes impact on gut microbial diversity, Gut microbiota, 2014 [Non-patent document 5] Tian H et al. J Clin Gastroenterol. 2015 [Non-patent document 6] Hecker MT et al. Open Forum Infect Dis. 2016 [Non-Patent Document 7] Prado et al. [Non-patent document 8] Dutch Blood Bank Association Guidelines (Section: Donor Exclusion Criteria) Summary of the Invention [Problem to be solved by the invention]

[0011] The above-described methods leave room for improvement in the treatment of cancer and related conditions, particularly cachexia. There remains a need to develop new or improved treatment strategies to improve patients' quality of life, increase life expectancy, reduce treatment side effects, reduce disease complications, and / or enhance response to treatment. Meeting this need is the purpose of the present disclosure. [Means for solving the problem]

[0012] We studied the effects of either fecal material transplantation (FMT) from an autologous (self) source or FMT from an allogeneic donor on recipient cancer patients, where the allogeneic donors had a range of body mass index (BMI), HOMA-IR, and age, while either had or did not have metabolic syndrome.

[0013] The inventors also investigated the effect of the allogeneic FMT on conditions that may be associated with cancer, particularly cachexia, which may be caused by the underlying cancer or may be a result of cancer treatment.

[0014] Surprisingly, the inventors have found that - At least 30 kg / m 2 Body Mass Index (BMI), and / or - HOMA-IR value of up to 2.5 mg / dL and / or age up to 60 years We have found that the use of allogeneic fecal material obtained from at least one donor subject with a tumor-associated phenotype results in increased overall survival, improved progression-free survival (PFS), a better response to or reduced toxicity / side effects of treatment (e.g., chemotherapy), and / or improved quality of life in the treatment of cancer or cachexia, and compared to autologous FMT, as well as compared to donors with a lower BMI and / or higher HOMA-IR values ​​and / or older age.

[0015] It was found that the PFS of patients with cancer-related cachexia after 6 months of receiving said allogeneic FMT was almost doubled compared to patients receiving autologous FMT. In addition, said use may lead to a reduction in (cancer-related) symptoms or complications and / or a reduction in (cancer) treatment-related symptoms or complications.

[0016] We found that the 30 kg / m 2No weight gain or increase in BMI was observed in recipients receiving the allogeneic fecal material compared to recipients receiving allogeneic fecal material from donors with a BMI below 100 and / or a HOMA-IR value above 2.5 and / or an age above 60 years. [Brief explanation of the drawings]

[0017] [Figure 1] 1 shows the mean resting energy expenditure (REE) for all subjects undergoing allogeneic fecal material transplantation (FMT) (line with highest REE at time 0) and all subjects undergoing autologous FMT (line with lowest REE at time 0). The shaded area indicates the standard deviation of the mean. [Figure 2] Figure 1 shows the mean percent body fat (measured by bioelectrical impedance analysis, BIA) for all subjects undergoing allogeneic FMT (lowest percent fat at time 0) and all subjects undergoing autologous FMT (highest percent fat at time 0). Shaded areas indicate the standard deviation of the mean. [Figure 3] Average appetite (measured by visual analog scale, VAS, questionnaire) for all subjects undergoing allogeneic FMT (line with lowest VAS score at time 0) and all subjects undergoing autologous FMT (line with highest VAS score at time 0). Shaded areas indicate standard deviation of the mean. [Figure 4] 1 shows the mean body mass index (BMI) for all subjects undergoing allogeneic FMT (line with lowest BMI at time 0) and all subjects undergoing autologous FMT (line with highest BMI at time 0). The shaded area indicates the standard deviation of the mean. [Figure 5] Responses of subjects who underwent allogeneic FMT (left three bars) and autologous FMT (right three bars). Each subject was classified into one of three categories: partial response, stable disease, and progression. [Figure 6] FIG. 1 shows progression-free survival probability over time for subjects undergoing allogeneic FMT (dashed line) and autologous FMT (solid line). [Figure 7]FIG. 1 shows survival probability over time for subjects undergoing allogeneic FMT (dotted line starting at 100%) and autologous FMT (dashed line starting at 100%). [Figure 8] FIG. 1 shows cumulative survival over a 1-year period for subjects undergoing allogeneic FMT (thick line with highest 1-year endpoint indicated by a "+" sign) and autologous FMT (non-thick line with lowest 1-year endpoint indicated by a "+" sign). [Figure 9] FIG. 1 shows cumulative survival over a 1-year period for subjects receiving allogeneic FMT from donor 23 (line with highest 1-year endpoint indicated by a "+" symbol), subjects receiving allogeneic FMT from all other donors (line with second highest 1-year endpoint indicated by a "+" symbol), and subjects receiving autologous FMT (line with lowest 1-year endpoint indicated by a "+" symbol). [Figure 10] Figure 1 shows cumulative survival over a 1-year period for subjects who underwent allogeneic FMT from donor 21 (line with highest 1-year endpoint indicated by a "+" sign), subjects who underwent allogeneic FMT from donor 23 (line with second highest 1-year endpoint indicated by a "+" sign), subjects who underwent allogeneic FMT from donor 14 (line with third highest 1-year endpoint indicated by a "+" sign), subjects who underwent allogeneic FMT from donor 22 (line ending at zero before 200 days), and subjects who underwent autologous FMT (line with lowest 1-year endpoint indicated by a "+" sign). DETAILED DESCRIPTION OF THE INVENTION

[0018] The present disclosure relates to the use of allogeneic fecal material for the treatment of cachexia and / or cancer, particularly cancer-associated cachexia, wherein the allogeneic fecal material is obtained from at least one donor subject, e.g., at least two, three, four donor subjects, wherein at least one donor subject is: - At least 30 kg / m 2 Body Mass Index (BMI), and / or - HOMA-IR value of up to 2.5 mg / dL and / or age up to 60 years It has.

[0019] Accordingly, the present disclosure relates to methods of treating cachexia or cancer, particularly cancer-associated cachexia, comprising administering to a patient in need thereof allogeneic fecal material as described herein.

[0020] Said treatment may significantly improve overall survival, progression-free survival (PFS), response to (chemo)therapy, quality of life, and / or reduce the toxicity / side effects of (chemo)therapy.

[0021] Survival time is the time between diagnosis of the disease and the time of death. Progression-free survival (PFS) is the length of time a patient survives with the disease during and after treatment for the disease and the disease does not progress. Disease progression is the development, spread, or worsening of the disease. Progression can continue until organ failure or patient death occurs. Therefore, PFS refers to the effectiveness of treatment because the disease (progression) is (temporarily) stopped.

[0022] Treatment according to the present disclosure is preferably not for weight gain and / or an increase in BMI. Transplantation of allogeneic fecal material according to the present disclosure typically does not result in an increase in the weight and / or BMI of the recipient.

[0023] According to another aspect, treatment according to the present disclosure is for improving quality of life, preferably reducing disease-related symptoms or complications and / or reducing (cancer) treatment-related symptoms or complications. Surprisingly, quality of life is significantly improved by transplantation of allogeneic fecal material according to the present disclosure.

[0024] Quality of life (QOL), as used herein, refers to the degree to which a person's objective needs are met in terms of self- or group-perception of subjective well-being. Changes in QOL can be quantified by comparing at least two responses to the same questions about happiness, life satisfaction, utility, and / or well-being at different points in time.

[0025] The term "donor," as used herein, refers to a subject who provides fecal material. Thus, fecal material according to the present disclosure can be obtained from a donor and administered to a recipient. The term "allogeneic," as used herein, means that the fecal material is not the patient's fecal material, but rather is derived from another subject of the same species, for example. Optionally, after processing, the fecal material is administered to the patient. Allogeneic is the opposite of "autologous," which means that the fecal material is the subject's own fecal material, i.e., obtained from the same subject and, optionally, after processing, administered to the same subject.

[0026] The term "body mass index" or "BMI" as used herein is obtained by dividing a person's weight by the square of the person's height and is expressed in kg / m 2 The value is expressed as 30 kg / m 2 Donors with a BMI greater than 31 kg / m are typically classified as obese. Preferably, at least one donor subject has a BMI of at least 31 kg / m 2 (average) body mass index (BMI) of at least 32 kg / m 2 and even more preferably at least 33 kg / m 2 (average) BMI of at least 34 kg / m 2 The fecal material of such donor subjects enhances progression-free survival and improves overall survival for cancer patients undergoing FMT using the fecal material.

[0027] The term "HOMA-IR" as used herein is an abbreviation for Homeostasis Model Assessment of Insulin Resistance. "HOMA-IR" refers to a value that represents an estimate of insulin resistance obtained by dividing the insulin and glucose levels in a person's blood. HOMA-IR values ​​can be calculated using the following formula:

[0028]

number

[0029] where H is the HOMA-IR value in mg / dL, glucose is the fasting glucose level in the blood in mmol / L, and insulin is the fasting insulin level in the blood in mIU / L. IU (for enzyme activity) is the abbreviation for International Unit, also called enzyme unit. Enzyme activity is the amount of substrate converted in one unit of time. 1 IU is equal to 1 μmol of substrate conversion per minute.

[0030] Preferably, and in accordance with the foregoing, at least one donor subject has a HOMA-IR value of less than 3.0, 2.9, 2.8, 2.7, 2.6, 2.5, or less than 2.4, 2.3, 2.2, 2.1, or 2.0 mg / dL, preferably less than 1.9, 1.8, or 1.7 mg / dL, more preferably less than 1.6, 1.5, 1.4, 1.3, 1.2, 1.1 mg / dL, and most preferably less than 1.0. Fecal material obtained from such donors has been found to increase the number of positive responders (i.e., cancer patients whose cancer has stabilized or not progressed).

[0031] Alternatively, or concurrently, at least one donor subject is preferably at most 60, 59, 58, 57, 56, 55, 54, 53, 52, 51, or 50 years old; more preferably at most 49, 48, 47, 46, 45, 44, 43, 42, or 41 years old; even more preferably at most 40, 39, 38, 37, 36, or 35 years old; even more preferably at most 34, 33, 32, 31, or 30 years old; and most preferably at most 29, 28, 27, 26, or 25 years old. Younger individuals often have relatively low HOMA-IR values. The inventors believe that fecal material from donors at most 60, 50, or 40 years old, for example, is more effective in treating cancer than fecal material obtained from older donors. The gut microbiota is a highly complex consortium of microorganisms that varies between individuals and over time. Therefore, the age of the donor's fecal material may influence the efficacy of allogeneic fecal material against the progression of cancer and related conditions. It is known that the microbiota of elderly people differs significantly from that of younger donors. See, e.g., Biagi et al. (PLoS One. 2010 May 17;5(5):e10667).

[0032] Preferably, at least one donor subject according to the present disclosure does not have metabolic syndrome. The absence of metabolic syndrome in a donor subject can further improve the efficacy of treatments using fecal material from such donor subjects, as it can further prolong overall survival, further enhance PFS, further reduce side effects and disease complications of (cancer) treatment, and enhance the quality of life of cachexia or cancer patients. Metabolic syndrome is present in approximately 50% of people classified as obese based on BMI. Surprisingly, this improvement can be observed by excluding people with metabolic syndrome from the donor group. The exclusion of donors with metabolic syndrome was not expected to have an effect on insulin resistance or insulin sensitivity in FMT recipients.

[0033] A person can be considered to have metabolic syndrome if three clusters of five conditions are considered to be interrelated. As defined by the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) and as used herein, the following five criteria: - Waist circumference greater than 40 inches or 102 cm (men) or 35 inches or 89 cm (women); - blood pressure above 130 / 85 mmHg, - Fasting triglyceride (TG) level greater than 150 mg / dL, - A fasting high-density lipoprotein (HDL) cholesterol level of less than 40 mg / dL (men) or less than 50 mg / dL (women), and - Fasting blood sugar level above 100 mg / dL If three or more of the following conditions are met, metabolic syndrome is present.

[0034] To measure a subject's fasting blood glucose (glucose) levels, fasting triglyceride levels, and fasting high density lipoprotein cholesterol levels in their blood, blood is drawn from the subject after the subject has not eaten or drunk anything other than water for at least 8 hours. Those skilled in the art are familiar with the methods used to quantify such levels.

[0035] At least one donor subject must have a blood glucose level of at least 30 kg / m 2In addition to having a body mass index (BMI) of 0.01 to 0.25, a HOMA-IR value of 0.01 to 2.5 mg / dL, and / or an age of 0.01 to 60 years, donors may be further selected by having a spinal subepithelial fat thickness of 0.01 to 0.25 mm, preferably 0.01 to 0.25 mm, more preferably 0.01 to 0.25 mm, and most preferably 0.01 to 0.25 mm. Transplantation of fecal material from donors with such spinal subepithelial fat thickness may further improve PFS, resulting in better chemotherapy response, reduced chemotherapy toxicity / side effects, and / or improved quality of life and overall survival in cancer patients. Thus, the preferred spinal subepithelial fat thickness of at least one donor subject may range from 14 to 36 mm, e.g., 16 to 34 mm, or 18 to 32 mm, or 20 to 30 mm, or 22 to 28 mm, or 24 to 28 mm.

[0036] Spinal subepithelial fat thickness is measured at the spinal subepithelial fat site, also known as the supraborhynchus site. The spinal subepithelial fat site is the intersection of two lines: (1) a line from the iliac spine to the edge of the anterior axilla, and (2) a horizontal line at the level of the iliac crest point. The iliac spine is the most inferior or lowest point of the tip of the anterior superior iliac spine (ASIS). The iliac crest point is the point on the iliac crest where the gland meets the ilium, drawn from the midaxillary (center of the armpit) along the longitudinal axis of the body.

[0037] Skinfold measurements can be taken using a skinfold caliper to determine the thickness of the skin at this location. A caliper is a device that can be used to measure the distance between opposing parts of an object. Measurements are typically taken on the right side of the body. By stretching and holding ("pinching") the skin at the spinal skinfold location, a double layer of skin and underlying adipose tissue is created with its outer edge in a longitudinal direction approximately parallel to the waist-to-head direction of the subject being measured. The caliper is applied 1 cm below the outer edge of the double skin layer at an angle perpendicular to the longitudinal direction of the skinfold. A reading in millimeters (mm) is taken 2 seconds after pinching the skinfold with the caliper. Spinal skinfold thickness is calculated as the average of two such skinfold measurements and associated readings.

[0038] Alternatively, or simultaneously, the subscapular skinfold thickness of at least one donor subject is preferably at most 45, 44, 43, 42, 41, or 40 mm, preferably at most 39, 38, 37, 36, or 35 mm, more preferably at most 34, 33, 32, 31, or 30 mm, and most preferably at most 29, 28, or 27 mm, e.g., 17-35 mm. Subscapular skinfold thickness measurements can be performed in a similar manner to the spinal skinfold thickness measurements described above, at a subscapular skinfold thickness site 2 cm (0.8 inches) along a line extending 45° outward and downward from the subscapular region when the donor subject is standing upright. The subscapular region is the lowest tip of the inferior angle of the scapula.

[0039] Other types of subcutaneous fat sites can be used in the present disclosure and are described, for example, in Applied Anatomy and Biomechanics in Sport, 2nd Edition, by T.R. Ackland, B. Elliott, and J. Bloomfield, 2009. Subcutaneous fat sites include, but are not limited to, a triceps skinfold site, the posterior part of the triceps at the level of the midacromio-radial landmark on the midline; a biceps skinfold site, the anterior most part of the biceps at the level of the midacromio-radial landmark; an abdominal skinfold site, 5 cm (2 inches) to the right of the navel or midpoint of the navel; an inner calf skinfold site, the site on the innermost aspect of the calf at the level of the greatest circumference; and an anterior thigh skinfold site, the site equidistant from the inguinal crease and the anterior surface of the patella on the midline of the thigh. Here, the acromion is the most superior point on the lateral edge of the acromion process when the subject is standing upright with their arms relaxed and hanging vertically, the radial point is the most basal point on the lateral edge of the radial head, and the midacromion-radial point is the point equidistant from the acromion and radial point landmarks.

[0040] It is within the scope of the present disclosure that subcutaneous fat thickness at any of the above subcutaneous fat sites or other subcutaneous fat sites can be measured to select at least one donor subject who can provide allogeneic fecal material for use in treatment according to the present disclosure, where allogeneic fecal material is defined herein.

[0041] At least one donor subject must have a blood glucose level of at least 30 kg / m 2 In addition to having a body mass index (BMI) of 0.01, a HOMA-IR value of 0.01 mg / dL or less, and / or an age of 0.01 mg / dL or less, a HOMA-IR value of 0.02 mg / dL or less, and / or an age of 0.02 mg / dL or less, the subject can be selected by having a thickness of 40 mm or less, preferably 32 mm or less, for example, 20 to 30 mm, at the triceps subcutaneous fat site. At the biceps subcutaneous fat site, the thickness can be 25 mm or less, preferably 18 mm or less, for example, 9 to 17 mm. At the medial calf subcutaneous fat site, the thickness can be 39 mm or less, preferably 32 mm or less, for example, 16 to 31 mm. At the abdominal subcutaneous fat site, the thickness can be 46 mm or less, preferably 38 mm or less, for example, 28 to 37 mm.

[0042] According to the present disclosure, at least 30 kg / m 2 In addition to having a body mass index (BMI) of 0.45-0.70 mm / m, a HOMA-IR value of at most 2.5 mg / dL, and / or an age of at most 60 years, at least one donor subject is preferably selected to have a waist circumference-to-height ratio of 0.45-0.70 mm / m, more preferably 0.50-0.65 mm / m, even more preferably 0.55-0.65 mm / m, and most preferably 0.58-0.62 mm / m. Thus, waist circumference is the donor subject's height in meters divided by their waist circumference in meters. Transplantation of fecal material from donors with such waist circumference-to-height ratios may further improve PFS, further enhancing recipient response to chemotherapy in cancer patients, reducing chemotherapy toxicity / side effects, and / or improving quality of life and / or prolonging overall survival.

[0043] Waist circumference, also known as abdominal circumference, as used herein, refers to the circumference of the waist measured at the level of the umbilicus. The umbilicus level is the height at which the umbilicus, also called the belly button or navel, is located. Waist circumference can be measured at the level of the umbilicus with a tape measure, the length of which is perpendicular to the longitudinal axis extending from the feet to the head of the subject whose waist circumference is being measured.

[0044] Alternatively, or simultaneously, preferably at least 30 kg / m 2 In addition to having a body mass index (BMI) of at most 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 for male subjects, and / or at most 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 for female subjects. While DEXA / DXA scans are commonly considered an accurate body fat test, those skilled in the art will appreciate other suitable methods for determining body fat percentage, such as bioelectrical impedance analysis (BIA).

[0045] In another aspect of the present disclosure, at least one donor subject preferably has a plasma creatine kinase level of at least 500 IU / L, preferably at least 650 IU / L, and most preferably at least 800 IU / L, which may result in increased overall survival, increased progression-free survival (PFS), better response to chemotherapy, reduced toxicity / side effects of chemotherapy, and / or improved quality of life for cancer and / or cachexia patients.

[0046] Creatine kinase, also known as phosphocreatine kinase or creatine phosphokinase (Enzyme Commission No. 2.7.3.2), catalyzes the reversible conversion of creatine to phosphocreatine. Normal plasma creatine kinase activity ranges from 60 to 400 IU / L.

[0047] Uses according to the present disclosure may include determining one or more of the BMI value, HOMA-IR value, age, waist circumference, waist circumference to height ratio, body fat percentage, fasting blood insulin level, fasting blood glucose level, blood triglyceride level, fasting high-density lipoprotein (HDL) cholesterol level and blood pressure, spinal subcutaneous fat thickness, subscapular subcutaneous fat thickness, and blood pressure of a pool of subjects, and then selecting one or more donor subjects as defined herein, for example, one or more donor subjects having the BMI, HOMA-IR, age, waist circumference, waist circumference to height ratio, body fat percentage, fasting blood insulin level, fasting blood glucose level, blood triglyceride level, fasting high-density lipoprotein (HDL) cholesterol level, blood pressure, spinal subcutaneous fat thickness, and / or subscapular subcutaneous fat thickness as defined herein.

[0048] The selected donor should, for example, have the following relevant characteristics: 30-40 kg / m 2 For example, 32 to 35 kg / m 2 BMI of 0.3 to 2.5 mg / dL, for example, 0.5 to 0.9 mg / dL; HOMA-IR of 18 to 60 years, for example, 20 to 40 years; waist circumference of 80 to 120 cm, for example, 95 to 105 cm; waist circumference to height ratio of 0.45 to 0.70 mm / m, for example, 0.55 to 0.65 mm / m; body fat percentage of less than 20% for male subjects and less than 25% for female subjects; and a maximum of 25 mIU / L, for example, 3 to 8 fasting blood insulin level of mIU / L; fasting blood glucose level of up to 130 mg / dL, e.g., 70-100 mg / dL; blood triglyceride level of up to 150 mg / dL, e.g., 50-150 mg / dL; fasting high-density lipoprotein (HDL) cholesterol level of up to 40 mg / dL for men and 50 mg / dL for women, e.g., 50-90 mg / dL for men and 60-90 mg / dL for women; spinal subcutaneous fat thickness of up to 36 mm, e.g., 20-30 mm; subscapular subcutaneous fat thickness of up to 40 mm, e.g., 20-32 mm; and / or blood pressure of up to 140 / 90 mmHg, e.g., 90 / 60 to 120 / 80 mmHg, or a combination thereof.

[0049] The selected donor subjects were 30–40 kg / m 2 Such donor subjects may have a BMI of 30-35 kg / m, a HOMA-IR value of 0.5-1.5 mg / dL, and / or an age of less than 30 years. 2 , e.g., 32-34 kg / m 2 The HOMA-IR values ​​of such donor subjects may range from 0.5 to 1.2 mg / dL, e.g., 0.5 to 0.9 mg / dL. The donor subjects may have an age of 18 to 30 years, e.g., 20 to 25 years.

[0050] Preferably, the donor subject weighs between 32 and 36 kg / m 2 The donor may have a BMI of 0.3 to 1.0 mg / dL, a HOMA-IR value of 0.3 to 1.0 mg / dL, and an age of 20 to 30 years. The spinal subepithelial fat thickness of such a donor may be 16 to 32 mm, e.g., 20 to 28 mm. The donor subject may have a very large abdominal circumference or a waist circumference greater than 102 cm, but may not have metabolic syndrome.

[0051] Preferably, the donor subject weighs between 32 and 36 kg / m 2 The donor may have a BMI of 0.3 to 2.0 or 0.3 to 1.0 mg / dL, a HOMA-IR value of 0.3 to 2.0 or 0.3 to 1.0 mg / dL, and an age of 20 to 30 years. The waist circumference to height ratio of such a donor may be 0.54 to 0.64 mm / m, for example, 0.56 to 0.62 mm / m. The donor subject may have a very large abdominal circumference or a waist circumference greater than 102 cm, but preferably does not have metabolic syndrome.

[0052] Fecal material according to the present disclosure can be feces, i.e., waste material passed from the intestine (anus), e.g., the (morning) stool, or portions thereof, and / or compositions derived therefrom. The fecal material can be purified, suspended in a medium, filtered, centrifuged, or otherwise processed, e.g., stabilized and lyophilized, to obtain a composition suitable for oral administration or administration in the gastrointestinal tract of a subject receiving treatment.

[0053] In one aspect of the present disclosure, the fecal material comprises a total of at least 5, 6, 7, 8, 9, 10, 11, 12 different phyla selected from the phyla Bacteria and Archaea, preferably at least 16 different phyla, more preferably at least 18 different phyla, and most preferably 20-34 different phyla, which may result in further increased overall survival, increased progression-free survival (PFS), better response to (chemo)therapy, reduced toxicity / side effects of (chemo)therapy, and / or improved quality of life for cancer and / or cachexia patients.

[0054] The total number of phyla can be determined by 16S rRNA amplicon sequencing as described in Clarke et al., Exercise and associated dietary extremes impact on gut microbial diversity, Gut microbiota, 2014.

[0055] The phyla are: Bacteria: Acidobacteria, Actinobacteria, Aquificae, Armatimonadetes, Bacteroidetes, Caldiserica, Chlamydiae, Chlorobi, Chloroflexi, Chrysioge Chrysiogenetes, Cyanobacteria, Deferribacteres, Deinococcus, Dictyoglomi, Elusimicrobia, Fibrobacteres, Firmicutes, Fusobacteria ), Gemmatimonadetes, Lentisphaerae, Nitrospira, Planctomycetes, Proteobacteria, Spirochaetes, Synergistetes, Tenericutes, Thermodesulfobacteria, Thermotogae and Verrucomicrobia, and / or the archaeal phyla: Crenarchaeota, Euryarchaeota, Korarchaeota, Nanoarchaeota and Thaumarchaeota. Preferably, fecal material according to the present disclosure comprises the phyla Bacteroidetes, Firmicutes, Proteobacteria, Actinobacteria, and / or Verrucomicrobia (and Euryarchaea).

[0056] In the context of this disclosure, cancer can be any type of cancer, including systemic and localized (organ-specific) cancer. The cancer can be bladder cancer, bone cancer, brain cancer, breast cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, gastro-esophageal cancer, head and neck cancer, liver cancer, lung cancer, lymphoma, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, stomach cancer, throat cancer, thyroid cancer, or cervical cancer.

[0057] In certain embodiments of the present disclosure, the cancer is selected from esophageal cancer, gastric cancer, gastroesophageal junction cancer, and pancreatic cancer. Patients with one of these types of such cancers would particularly benefit from the allogeneic fecal material described herein, for example, obtained from at least one donor subject, wherein the at least one donor subject is - At least 30 kg / m 2 Body Mass Index (BMI), and - HOMA-IR value of 2.5 mg / dL or less and / or age of 60 or 40 years or less It has.

[0058] The average overall survival of patients with such types of cancer who undergo FMT from an allogeneic donor as described herein may be improved by 25% compared to patients who undergo FMT from an autologous donor. In addition, fecal material obtained from donors with a BMI greater than 25, e.g., greater than 30, and a HOMA-IR value less than 2.5, e.g., less than 1.0, and an age less than 40 years, is particularly suitable for improving overall survival and progression-free survival of patients with such types of cancer.

[0059] Esophageal cancer, or esophageal cancer, originates in the cells of the esophagus, a long, hollow tube that connects the throat to the stomach. The esophagus moves swallowed food to the stomach. Esophageal cancer causes tumors to form within the esophagus, obstructing this movement by narrowing the esophageal tube. Common symptoms of esophageal cancer include difficulty swallowing, hoarseness, and pain in the area around the stomach. Esophageal cancer can be classified into two main types: esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). Risk factors for ESCC include smoking, alcohol use, and a poor diet. For EAC, risk factors include smoking, acid reflux, and obesity.

[0060] Esophageal cancer has a poor prognosis, with only 13-18% of patients surviving five years after diagnosis. An important reason for this is the relatively late diagnosis, as symptoms often only appear when esophageal cancer is at an advanced stage.

[0061] Gastric cancer, also known as stomach cancer, originates in the cells of the stomach. The stomach is a hollow digestive organ located between the esophagus and the duodenum. Gastric acid and various enzymes are secreted by the stomach to aid in food digestion. Common symptoms of gastric cancer include loss of appetite, nausea and vomiting, acid reflux, melena, and weight loss. Risk factors for gastric cancer include Helicobacter pylori infection, smoking, poor diet, and obesity. The prognosis for gastric cancer is also very poor. Approximately 32% of patients survive five years after diagnosis. As with esophageal cancer, symptoms most often occur when the cancer has reached an advanced stage, contributing to the poor prognosis.

[0062] Gastroesophageal junction cancer arises in glandular cells located near the gastroesophageal junction in the lower part of the esophagus where it connects to the stomach. Common symptoms of gastroesophageal cancer include painful swallowing, weight loss, and nausea and vomiting. Gastroesophageal cancer is considered a type of esophageal cancer and is treated similarly. Risk factors include obesity, smoking, alcohol use, and a poor diet. As with esophageal and gastric cancers, the 5-year survival rate for gastroesophageal cancer varies greatly depending on the stage of the cancer at the time of diagnosis, ranging from 5 to 43%.

[0063] Pancreatic cancer originates in the cells of the pancreas, an organ of the digestive system located near the stomach that regulates blood sugar levels and neutralizes acid that enters the duodenum. The most common type of pancreatic cancer is pancreatic adenocarcinoma. Common symptoms of pancreatic cancer include loss of appetite, weight loss, yellowing of the skin, and back pain. Risk factors include smoking, diabetes, and obesity. The prognosis for pancreatic cancer is very poor, with an average five-year survival rate of only 6%.

[0064] According to the present disclosure, the cancer can be metastatic cancer. Metastatic cancer is cancer that has spread locally by migrating from the primary tumor to nearby tissues. For many types of cancer, metastatic cancer is referred to as stage IV cancer. The process of cancer spread is called metastasis. Metastatic cancer is more difficult to treat compared to localized cancer. Palliative care is more frequently applied in metastatic cancer. The inventors have found that allogeneic FMT significantly improves quality of life (QoL) in cases of metastatic cancer.

[0065] The cancer type can be selected from adenocarcinoma, squamous cell carcinoma, and undifferentiated carcinoma. Adenocarcinoma (AC) can occur in various parts of the body. It is a type of excessive and abnormal proliferation of epithelial cells (glandular cells) that synthesize substances to be released anywhere in the body. Adenocarcinoma is a common type of esophageal cancer, pancreatic cancer, gastric cancer, and gastroesophageal cancer.

[0066] Squamous cell carcinoma arises in squamous cells. These cells look like polygonal plates when viewed from above. These cells are located near the surface of tissues within body cavities, allowing compounds to diffuse through these cells (surface cells). Squamous cell carcinoma is a common type of esophageal cancer.

[0067] A cancer is undifferentiated if it does not resemble any normal glandular or surface cells. Undifferentiated cancers have a very poor prognosis and a more aggressive clinical course than differentiated cancers, such as adenocarcinoma and squamous cell carcinoma.

[0068] In a further aspect of the present disclosure, cancer is accompanied by cachexia and / or sarcopenia. The present disclosure reduces the deleterious effects of cachexia on the body, resulting in increased overall survival, increased progression-free survival (PFS), better response to chemotherapy, reduced toxicity / side effects of chemotherapy, and / or improved quality of life.

[0069] Cachexia can be described as a state of irreversible wasting, i.e., progressive weakness and emaciation of the body, which may be associated with cancer. Age-related physiological muscle wasting and debilitation is known as sarcopenia. Sarcopenia can occur in conjunction with cachexia in elderly people. In cachexia, skeletal muscle and adipose tissue are used for body nutrition, reducing the ability of cancer patients to tolerate cancer treatment with chemotherapy and radiotherapy. Generally, cancer patients with cachexia die if their weight loss exceeds 25-30%.

[0070] The present disclosure also relates to allogeneic fecal material as described herein for the treatment of cachexia, sarcopenia and / or anorexia nervosa, e.g., the allogeneic fecal material is obtained from at least one donor subject, the at least one donor subject being a subject as described herein, e.g., - At least 30 kg / m 2 Body mass index (BMI), - HOMA-IR value of up to 2.5 mg / dL and / or age up to 60 years It has.

[0071] Congestive heart failure, chronic obstructive pulmonary disease, chronic kidney disease, and AIDS may also be complicated by cachexia and sarcopenia, and are therefore encompassed by the present disclosure. Anorexia nervosa is also encompassed by the present disclosure and can be characterized by weight loss, dietary restriction, fear of weight gain, and a strong desire to be thin. Diagnostic criteria (DSM-5) include restriction of energy intake relative to needs that results in underweight, an intense fear of weight gain, or persistent behaviors that prevent weight gain, and the person experiencing weight or shape disturbances or lack of awareness of the risk of being underweight.

[0072] Allogeneic fecal material according to the present disclosure can be administered to a subject's small intestine, preferably the duodenum. Administration to the duodenum is beneficial because the fecal material is not affected by other parts of the gastrointestinal tract upstream of the duodenum, such as the stomach. The duodenum is the first and shortest section of the small intestine, preceding the jejunum and ileum. In humans, the duodenum is a 25-38 cm hollow tube that connects the stomach to the distal duodenum. It begins at the duodenal bulb and ends at the duodenal levator muscle.

[0073] Preferably, the allogeneic fecal material for use in accordance with the present disclosure is feces or a portion thereof, preferably a purified portion thereof. Purifying the fecal material allows the fecal material to be more conveniently introduced into the duodenum of the subject receiving treatment. The presence of less debris in purified fecal material enhances the flow rate through the probe used to administer the fecal material and reduces the thickness of the probe, thereby reducing discomfort to the subject receiving treatment.

[0074] Portions of fecal matter, as used herein, refers to one or more specific groups of components including, but not limited to, enzymes, proteins, lipids, molecules, microorganisms, viruses, bacteria, fungi, yeasts, archaea, compounds, complexes, solids, liquids, particles, and fibers.

[0075] A purified portion of fecal material, as used herein, means that no undesired components are present in the fecal material.

[0076] Preferably, allogeneic fecal material for use according to the present disclosure is contained in a liquid medium and / or does not contain solids having a diameter greater than 10, 25, 50, 75, 100, 200, 400, 600, 800, or 1000 μm, and is preferably obtained by mixing the allogeneic feces with an aqueous medium followed by filtration and / or centrifugation. This greatly reduces viscosity and enhances the flow rate of the fecal material, facilitating administration of the fecal material to the gastrointestinal tract or duodenum of the subject being treated. The liquid medium may include water or other types of liquids, to which other components, such as salts, may be added to make the solution isotonic.

[0077] Preferably, the allogeneic fecal material for use according to the present disclosure is mixed with a chemotherapeutic agent, preferably selected from the group consisting of capecitabine or oxaliplatin. This combination with a chemotherapeutic agent can be highly effective in treating cancer, as it prolongs overall survival, cumulative survival, enhances PFS, and reduces the side effects of cancer and cancer treatment.

[0078] Capecitabine is a chemotherapy drug used to treat various types of cancer, including esophageal, gastric, and breast cancer. It is converted in the body to 5-fluorouracil, a thymidylate synthase inhibitor. Thymidylate synthase is required for de novo DNA synthesis. Because DNA synthesis is required for cell division, capecitabine inhibits the formation of cancer cells.

[0079] Oxaliplatin is a platinum-containing chemotherapy drug that can be used in conjunction with capecitabine. Oxaliplatin is cytotoxic and causes cell death by cross-linking DNA, preventing its replication and transcription. Oxaliplatin is often used to treat colorectal cancer.

[0080] According to one aspect of the present disclosure, the fecal material according to the present disclosure is contained in a composition, preferably a pharmaceutical composition, more preferably a liquid or solid dosage form, most preferably a capsule, tablet, or powder, to facilitate administration of the fecal material to a recipient.

[0081] It is further contemplated that fecal material according to the present disclosure will be present in a freeze-dried and / or microencapsulated form, which will allow for the preservation of the fecal material and allow for its storage for extended periods of time, for example, months or years.

[0082] Uses according to the present disclosure preferably comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 separate administrations of fecal material obtained from at least one donor subject to, for example, the small intestine, preferably the duodenum, of a recipient, preferably with an interval of at least 1, 2, 3, 4, 5, 6, 7, 8 weeks between said separate administrations.

[0083] According to the present disclosure, fecal material may be administered enterally, preferably orally, nasally or rectally, and / or via a nasoduodenal tube.

[0084] For oral administration, fecal matter, such as one or more components of autologous feces, preferably in lyophilized form (see, for example, Tian H et al., J Clin Gastroenterol. 2015, or Hecker MT et al., Open Forum Infect Dis. 2016), can be administered in solid dosage forms such as capsules, tablets, and powders, or in liquid dosage forms such as elixirs, syrups, and suspensions. Additionally, carriers such as activated carbon can be used.

[0085] The fecal material may be used as a medicine and / or accompanied by a physiologically acceptable carrier, which may be any inert carrier. For example, non-limiting examples of suitable physiologically or pharmaceutically acceptable carriers include any well-known physiological or pharmaceutical carriers, buffers, diluents, and excipients. The selection of a suitable physiological carrier will obviously depend on the intended method of administration of the compositions taught herein (e.g., oral) and the intended form of the composition (e.g., beverage, yogurt, powder, capsule, etc.). Those skilled in the art will know how to select a physiologically acceptable carrier that is suitable for or compatible with the compositions for use as taught herein.

[0086] The fecal material is expected to be contained and / or encapsulated in an (enteric) coating, where preferably the coating does not dissolve and / or disintegrate in the subject's stomach environment. Such a coating can help the fecal material reach the intended delivery site, e.g., the duodenum, without being degraded by the acidic environment of the stomach. A preferred (enteric) coating works by presenting a surface that is stable at the highly acidic pH found in the stomach but degrades more rapidly at lower pH. For example, it will not dissolve in the gastric acid of the stomach (pH approximately 3), but will dissolve in the alkaline (pH 7-9) environment present in the small intestine or duodenum.

[0087] In this document and in the claims, the verb "comprise" and its conjugations are used in an open-ended sense, meaning to include items that follow the word, but not to exclude items not specifically mentioned. In addition, when referring to an element with the indefinite article "a" or "an," it does not exclude the possibility that more than one element is present, unless the context clearly requires that only one element be present. Thus, the indefinite article "a" or "an" typically means "at least one." [Example]

[0088] A randomized, double-blind, placebo-controlled interventional trial was designed to include 16 patients with metastatic or locally advanced esophageal or gastric cancer who had received standard first-line palliative chemotherapy. The treatment consisted of 3-weekly cycles of capecitabine (1000 mg / m 2 , twice daily on days 1 to 14) and oxaliplatin (130 mg / m on day 1 2 ) for up to six consecutive cycles followed by capecitabine maintenance therapy (Table 1).

[0089] [Table 1]

[0090] Screening and Inclusion A screening visit was conducted at the AMC (Academisch Medisch Centrum, Amsterdam). During this screening, inclusion and exclusion criteria were verified, and informed consent was obtained after a verbal and written explanation of the study. Medical history was recorded, and a physical examination was performed, including weight, height, waist and hip circumference, and blood pressure. If patients were found to be eligible for the study, they were randomized into two treatment groups: 1. Autologous fecal transplant (n=12) 2. Allogeneic fecal transplantation from obese subjects (n=12)

[0091] Study visits and measurements Two follow-up study visits were conducted at baseline and 4 weeks later (the 12-week visit was conducted in conjunction with regular outpatient visits). Subjects fasted overnight before each visit. The total volume of blood obtained during this study was 180 ml: 60 ml at weeks 0, 4, and 12 (the last 12 weeks were collected in the outpatient clinic in conjunction with regular blood collection during regular treatment follow-up). Further imaging for muscle mass measurement (sarcopenia) was not required for this study, as it is standard procedure to perform CT scans to assess treatment response.

[0092] Study Visits: Baseline, 4 and 12 Weeks Before both visits, subjects collected a 48-hour fecal sample, a morning stool sample, and a 24-hour urine sample. Resting energy expenditure (REE) and bioelectrical impedance analysis (BIA) were assessed. Additionally, subjects were weighed to determine BMI, and blood samples were collected (60 ml per visit). Chemotherapy toxicity was assessed and graded according to the Common Terminology Criteria for Adverse Events (CTCAE) while completing a hunger questionnaire. At baseline, fecal transplantation was performed via duodenal tube placement (using coretrack). After duodenal tube placement, fecal infusion (autologous or allogeneic, depending on randomization) was performed. On the day before fecal transplantation, bowel cleansing was performed by drinking 1 liter of macrogol after dinner (7 p.m.–midnight).

[0093] Study Visit Donors: Baseline At baseline, donors collected a 48-hour fecal sample, a morning fecal sample (for transplant), and a 24-hour urine sample. Initially, resting energy expenditure (REE) and bioelectrical impedance analysis (BIA) were assessed. A questionnaire about hunger levels was completed. Fresh morning feces was used for each donation.

[0094] Inclusion criteria To be eligible to participate in this study, patients must meet all of the following criteria: - Men or women with metastatic or locally advanced esophageal and / or gastric cancer who have received standard first-line palliative chemotherapy (capecitabine / oxaliplatin) - Ages 30-70 - Sarcopenia criteria were met using computed tomography (CT) scans: L3 muscle area surface was normalized to patient height to calculate L3 muscle index, cm 2 / m 2 The cutoff value used for sarcopenia was determined by Prado et al. 2 Based on the method, for men it is 52.4 cm 2 / m 2 and 38.5cm for women. 2 / m 2 is. - International Classification of Functioning, Disability and Health (ICF) 3 , meet WHO 1, 2 or 3 - Stable medication use, all subjects use PPIs. - Subjects must be able to give informed consent and must do so willingly.

[0095] Exclusion criteria Potential subjects who meet any of the following criteria will be excluded from participation in this study: - Smoking, XTC, amphetamine or cocaine abuse - Alcohol abuse (>3 / day) - Cholecystectomy - HIV infection with CD4 count <240 - Chronic nausea, altered taste, difficulty swallowing, or mechanical obstruction due to malignant tumors - History of neurological or psychiatric disorders - Patients with diabetes mellitus (several studies have shown that high levels of NLR reflect ongoing vascular inflammation and play an important role in the pathophysiology of DM and also in prediabetes) 4

[0096] Subject screening Starting with 46 subjects, 3 had diabetes mellitus, 1 received antibiotics, 2 were not treated with CAPOX, and 2 had an International Classification of Functioning, Disability and Health (ICF) score. 3 ) were excluded because they did not meet the criteria. Of the remaining 38 subjects, 10 subjects were excluded because the study was too stressful or burdensome for them, and another 4 subjects were excluded because the distance to travel was extensive. The remaining 24 patients were randomized into two groups. Twelve patients were placed in the autologous transplant group, and the remaining 12 patients were placed in the allogeneic transplant group. Table 2 below shows the characteristics of these patients in both groups (autologous (n=12) and allogeneic (n=12)). Here, n refers to the number of subjects, the brackets include the minimum and maximum values ​​(age or BMI), and the percentage between the brackets indicates the number of subjects in a particular class (male or female) divided by the total number of subjects in one of the groups (autologous or allogeneic), expressed as a percentage.

[0097] [Table 2]

[0098] Of the 24 subjects included, 8 were excluded during the study. Of these 8 subjects, 3 received antibiotics during the study, 3 died, and 4 discontinued the study. Ultimately, 16 subjects completed the study.

[0099] Donor Inclusion Criteria To be eligible to participate as a stool donor in this study, subjects must meet all of the following criteria: - Caucasian male or female - Ages 18-70 - BMI > 30 kg / m 2

[0100] Donor Exclusion Criteria Potential donors who meet any of the following criteria will be excluded from participating as donors in this study: - Presence of chronic low-grade inflammation or criteria for metabolic syndrome - Use of any medications, including PPIs and antibiotics - Presence of type 2 diabetes or hypertension - diarrhea - Cholecystectomy - HIV, HAV, HBV, HCV, active CMV, active EBV, IBD - Risky sexual behavior (survey) - Presence of fecal bacterial pathogens (Salmonella, Shigella, Campylobacter, Yersinia) or parasites - C. difficile positive stool test

[0101] Individuals with increased risk for one of the above conditions (homosexual contact, recent blood transfusion) are excluded, and donors are not recruited within the healthcare provider.

[0102] Sample size calculation Sample size calculations were based on visual analog scale (VAS) results for satiety (primary endpoint) and appetite (secondary endpoint). This was based on an expected mean increase in VAS score of 15 mm (SD 10 mm) for allogeneic lean donor FMT versus a 5 mm increase for autologous FMT. Considering a two-sided p-value (alpha) of 0.05 and 80% power, a total of 16 subjects were required. Ideally, to assign cancer patients to one healthy obese donor, up to eight donors would be required for stool donation. An interim analysis of these 16 patients was performed, and the power calculation was repeated.

[0103] Primary Study Parameters / Endpoints - Satiety, measured by visual analog scale (VAS) questionnaire - Body composition measured by bioelectrical impedance analysis (BIA) - Treatment response, measured by CT scan at baseline and after the first 3 cycles of chemotherapy (week 12) - Overall survival (defined as the number of days alive after PA diagnosis)

[0104] Randomization, blinding, and treatment assignment Patients were randomized using a computer program (ALEA). Blinding was ensured in both groups by mixed fecal transplants. On the day of fecal transplant, both subjects and donors provided morning feces. Randomization and preparation of the feces were performed by one of the research assistants. He / she was the only person who knew the treatment the subjects had received and their further participation in the study. The feces was placed in a 500 ml glass bottle. This appeared as a brown liquid, unrecognizable as feces, from the donor or subject and was handed over to the researcher who performed the nasoduodenal tube instillation. Subjects, donors, and researchers were all blinded. Sufficient experience (>300 fecal transplants in the last 8 years at AMC) has been gained with this procedure.

[0105] Stool donor screening Potential donors were thoroughly screened according to the protocol used in our previously approved FAECAL & FATLOSE trial (MEC 07 / 114 and 11 / 023) by our METC. Donor feces were collected from obese donors who were screened according to the Dutch Blood Bank Association guidelines (section: Donor Exclusion Criteria). Donors underwent indirect calorimetry (to generate energy expenditure) and bioimpedance analysis (BIA) as well as blood sampling (60 ml).

[0106] Preparation of fecal enema for fecal transplant On the day of infusion, donors and recipients provided fresh fecal samples (average 150-250 grams) (generated within 24 hours prior to use). After collection in a plastic flask, the feces were covered with saline and stored at room temperature. Collection times were recorded. All procedures were performed in the Clinical Microbiology Department at AMC. All steps were performed in a fume hood by experienced laboratory colleagues or researchers. The fresh fecal solution was mixed for 10 minutes until completely homogenized. After this, the fecal solution was poured and strained through a clean metal sieve to remove large food debris. This process was repeated. The homogenized solution was then decanted through a clean metal funnel into a 1000 ml sterile glass bottle. Samples of the processed fecal infusion solution were then collected for later analysis, and the bottles were kept at standard room temperature (17°C) until the patient completed their bowel cleansing.

[0107] Colon cleansing The bowel was cleansed (according to standard protocols) the night before fecal transplant with 3–4 liters of macrogol (Klean Prep) depending on the subject's weight (4 liters for >60 kg, 3 liters for <60 kg) to ensure complete bowel cleansing (duration 3–4 hours).

[0108] fecal injection Finally, the filtered and mixed fecal transplant (<6 hours after processing) was instilled into the duodenum via a duodenal tube.

[0109] Resting Energy Expenditure (REE) REE was measured by indirect calorimetry at baseline and after 4 weeks. Oxygen consumption and CO2 production were measured for 20 minutes using a fume hood system. From these measurements, REE and respiratory quotient (RQ) were calculated. RQ represents the ratio of carbon dioxide exhaled by an individual to the amount of oxygen consumed, and represents whole-body substrate oxidation (glucose, lipid, and protein oxidation). 24-hour energy expenditure was estimated using Weir's simplified formula.

[0110] No significant differences in the REE of subjects between the allogeneic and autologous groups were observed over 12 weeks (Figure 1). Additionally, no significant differences in the REE of subjects within each group were observed over time. Therefore, FMT does not appear to have an effect on the REE of subjects receiving FMT.

[0111] Bioelectrical Impedance Analysis (BIA) Body composition was measured at baseline and after 4 and 12 weeks using BIA. BIA determines the electrical impedance of current through body tissues, which can then be used to calculate and estimate body fat. BIA is a safe, non-invasive, and inexpensive clinical method for assessing body composition. 5 .

[0112] No significant differences in the fat percentage of subjects between the allogeneic and autologous groups were observed over 12 weeks (Figure 2). In addition, no significant differences in the fat percentage of subjects within each group were observed over time. Therefore, FMT does not appear to have an effect on the fat percentage of subjects undergoing FMT.

[0113] Appetite assessment (VAS questionnaire) At baseline and 4 and 12 weeks post-transplant, subjects were also asked to complete the following questionnaire (next to the satiety VAS) to measure appetite levels: Visual Analogue Scale (VAS): Measurement of appetite levels.

[0114] There appears to be no effect of FMT on appetite as measured by the visual analog scale (Figure 3). No significant difference in the appetite of subjects between the allogeneic and autologous groups was observed over 12 weeks (Figure 3). In addition, no significant difference in the appetite of subjects within each group was observed over time. Therefore, FMT appears to have no effect on the appetite of subjects who underwent FMT.

[0115] BMI measurement of subjects The subjects' BMI was measured at the start of the study (baseline) and at weeks 4 and 12. No significant changes in the BMI of subjects in both groups (allogeneic and autologous) were observed over time, as can be seen in Figure 4.

[0116] Treatment success measured by CT scan Subjects' responses to FMT were measured by CT scan at baseline and after the first three cycles of chemotherapy (week 12). Responses were classified as partial response, stable disease, and progression using the Response Evaluation Criteria in Solid Tumors (RECIST). Compared with the group receiving allogeneic FMT, progression was significantly higher in the group receiving autologous FMT, and partial responses were lower in this group (Figure 5).

[0117] Progression-free survival The median progression-free survival for subjects in the autologous FMT group was 139 days, while the median progression-free survival for subjects in the allogeneic FMT group was 243 days (Figure 6). The difference of 104 days (p=0.11) indicates that allogeneic FMT has a significant effect on progression-free survival. Table 3 below shows the progression-free survival (PFS) duration and corresponding percentages for subjects in both groups.

[0118] [Table 3]

[0119] Cox regression was used to study the difference in the hazard of cancer progression between subjects receiving autologous and allogeneic FMT. A Cox hazard ratio of 0.497 (P = 0.13) was obtained, indicating that subjects in the allogeneic group had a 49.7% lower hazard of cancer progression compared with subjects in the autologous group.

[0120] overall survival The median survival time for subjects in the autologous FMT group was 331 days, and the median survival time for subjects in the allogeneic FMT group was 414 days (Figure 7). Thus, the median survival time for subjects in the allogeneic group improved by 83 days (log-rank test P=0.22). The 6-month survival probability for the autologous group was 36.6%, compared with 91.6% for subjects in the allogeneic group. The 1-year survival probability (still ongoing) was 32.7% for subjects in the autologous group, compared with 58.3% for subjects in the allogeneic group.

[0121] The Cox hazard ratio was 0.561, indicating that subjects undergoing allogeneic FMT had a 56.1% lower hazard of death compared to subjects undergoing autologous FMT.

[0122] Quality of life Esophageal cancer survival is extremely poor, with most patients dying within one year of diagnosis. Therefore, the 104-day improvement in progression-free survival for subjects receiving allogeneic fecal material transplantation (FMT) compared with subjects receiving autologous FMT is a highly significant effect. In this regard, the 83-day improvement in median survival also supports allogeneic FMT. Unexpectedly, appetite did not improve in subjects receiving fecal material from obese donors (Figure 3).

[0123] In addition to improved survival, the quality of life of subjects undergoing allogeneic FMT is significantly improved. A primary goal of palliative care is optimizing quality of life. This goal is particularly achieved by the present disclosure because symptoms of cachexia are reduced in subjects undergoing allogeneic FMT. [Example]

[0124] Allogeneic fecal material transplantation (FMT) was most effective when the fecal material donor had a BMI of at least 30. However, differences in FMT efficacy between donors within this allogeneic group appear to exist. This effect was further examined.

[0125] FMT donor Within the group of patients who underwent allogeneic FMT, there were four different donors (donors 14, 21, 22, and 23). Some characteristics of these donors are listed in Table 4.

[0126] [Table 4]

[0127] The donations listed in Table 4 indicate the number of patients in the allogeneic FMT group who received FMT from the indicated donor. For example, four patients received FMT from donor 22. The response of patients in the allogeneic group to FMT from the above donors is shown in Table 5.

[0128] [Table 5]

[0129] Of the 12 patients in the autologous FMT group, 7 experienced disease progression, while 5 experienced stable disease or a partial response. In contrast, only 2 of the 12 patients in the allogeneic FMT group experienced disease progression, while the vast majority of patients, 10, experienced stable disease or a partial response.

[0130] cumulative survival Cumulative survival is the proportion of patients who survive beyond a given period of time. The cumulative survival of patients in both groups (autologous and allogeneic FMT groups) after one year is shown in Figure 8.

[0131] As shown in Figure 8, patients who underwent allogeneic FMT had a higher cumulative survival at 1 year compared to patients who underwent autologous FMT.

[0132] In Figure 9, the cumulative survival of patients who received allogeneic FMT from donor 23 (the line with the highest 1-year endpoint) is compared with all other patients in both the allogeneic and autologous groups (the line with the lowest 1-year endpoint) and with all other patients in the allogeneic FMT group (the line between the other two lines with intermediate endpoints). Thus, a significantly positive effect of donor 23-derived FMT can be observed (Figure 9).

[0133] Figure 10 shows the cumulative survival of patients for each specific donor compared with the autologous FMT group. As shown, only one patient who received FMT from donor 21 (see Table 5) was still alive one year later (highest one-year endpoint line), while the patient who received FMT from donor 22 died before the one-year mark. Four of six patients who received FMT from donor 23 were still alive one year later (second-highest one-year endpoint line), and two of four patients who received FMT from donor 14 were still alive one year later (third-highest one-year endpoint line). Only two of 12 patients who received autologous FMT were still alive one year later.

[0134] Factors influencing the efficacy of allogeneic FMT Therefore, there are likely donor-specific aspects that influence the efficacy of allogeneic FMT. Most preferably, the BMI of the FMT donor is greater than 30. Because BMI is not a measure that distinguishes between donor 23 and donors 21 and 22, we quantified the donor's insulin resistance.

[0135] Insulin resistance (IR) and beta cell function (β) can be quantified by homeostasis model assessment (HOMA). Fasting glucose and insulin levels can be used to estimate insulin resistance (HOMA-IR) and beta cell function (HOMA-β). In the group of patients undergoing allogeneic FMT, all six patients who underwent FMT from donor 23 responded. Donor 23 was a healthy young rugby player, whereas donors 14 and 21 were older and likely had higher HOMA-IR values. Donor 23's HOMA-IR value was less than 2.5, whereas donors 14 and 21 both had HOMA-IR values ​​greater than 2.5.

[0136] To diagnose obesity in athletes such as rugby players, body fat can be assessed by skinfold measurements or by measuring waist circumference to height ratio. An estimated 72% of athletes are misclassified as obese due to their BMI.

[0137] Additionally, donor 23 is much younger than donors 21 and 22. Therefore, there is likely an effect of donor age on the outcome of FMT. Preferably, the donor's age is less than 40. Based on donor 23, in addition to the factors mentioned above, there is likely a beneficial effect for patients undergoing FMT from donors with low body fat and high muscle strength. [Example]

[0138] Administration of allogeneic fecal material from various donors to recipients diagnosed with various types of cancer Patients (recipients) with the cancer types indicated below will receive allogeneic fecal material via duodenal tube administration (as in Example 1) after bowel cleansing according to the following three treatment groups: 1. 18-30 kg / m to recipient at 0, 8, and 16 weeks 2 Transplantation of allogeneic fecal material from a donor with a BMI of 2. At least 30 kg / m2 to the recipient at 0, 8, and 16 weeks 2Transplantation of allogeneic fecal material from a donor with a BMI of 2a. Allogeneic fecal material transplant in treatment arm 2, where the donor has a HOMA-IR value of less than 2.5 mg / dL. 2b. Allogeneic fecal material transplant in treatment arm 2, where the donor is 40 years of age or younger. 2c. Allogeneic fecal material transplant in treatment group 2a, where the donor does not have metabolic syndrome. 2d. Allogeneic fecal material transplant in treatment group 2c, where the donor has a spinal subepithelial fat thickness of less than 36 mm and a waist circumference-to-height ratio of 0.50-0.65 mm / m.

[0139] [Table 6]

[0140] Similar results for PFS and QOL may be obtained for recipients with cachexia or sarcopenia, with or without a cancer diagnosis. Furthermore, it is expected that results similar to the estimated effects shown in Table 6 above may be obtained with a larger patient cohort.

[0141] [References] TIFF0007817952000008.tif102159

Claims

1. 1. A composition comprising allogeneic fecal material for use in treating cancer patients with cachexia, wherein the allogeneic fecal material is obtained from at least one donor subject, the at least one donor subject comprising: - At least 30 kg / m 2 Body Mass Index (BMI), and - HOMA-IR up to 2.5 mg / dL and / or age up to 60 years A composition comprising:

2. At least one donor subject had a blood glucose level of at least 31 kg / m 2 The composition of claim 1, having a BMI of

3. 3. The composition of claim 1 or 2, wherein at least one donor subject has a HOMA-IR of up to 2.0 mg / dL.

4. 4. The composition of any one of claims 1 to 3, wherein at least one donor subject has an age of up to 40 years.

5. 5. The composition of any one of claims 1 to 4, wherein at least one donor subject does not have metabolic syndrome.

6. The use comprises determining one or more of BMI, HOMA-IR, age, metabolic syndrome of a pool of subjects; and Subsequent selection of one or more donor subjects as defined in any one of claims 1 to 5.

6. The composition of any one of claims 1 to 5, comprising:

7. The composition of claim 1, wherein the cancer is selected from esophageal cancer, gastric cancer, gastroesophageal cancer, and pancreatic cancer.

8. 8. The composition of any one of claims 1 to 7, wherein the use is for the prolongation of survival.

9. 9. A composition according to any one of claims 1 to 8, wherein the use is for improving response to chemotherapy and / or reducing the side effects of chemotherapy.

10. 10. The composition of any one of claims 1 to 9, wherein the use is not for weight gain and / or increasing BMI.

11. 11. The composition of any one of claims 1 to 10, wherein the use is for improving quality of life.

12. 12. The composition of claim 11, wherein the use is for the reduction of disease-related symptoms or complications and / or the reduction of treatment-related symptoms or complications.

13. 13. The composition of any one of claims 1 to 12, wherein the allogeneic fecal material is administered to the small intestine of the recipient subject.

14. 14. The composition of any one of claims 1 to 13, wherein the allogeneic fecal material is feces or a portion thereof.

15. 15. The composition of claim 14, wherein the allogeneic fecal material is a portion of purified feces.

16. 16. A composition according to any one of claims 1 to 15, wherein the fecal material is contained in a liquid medium and / or does not contain solids having a diameter greater than 10, 25, 50, 75, 100, 200, 400, 600, 800 or 1000 μm.

17. 17. The composition of any one of claims 1 to 16, wherein the fecal material is combined with a chemotherapeutic agent.

18. 18. The composition of claim 17, wherein the chemotherapeutic agent is selected from the group consisting of capecitabine or oxaliplatin.

19. 19. The composition of any one of claims 1 to 18, wherein the allogeneic fecal material is comprised in a pharmaceutical composition.

20. 20. The composition of claim 19, wherein the allogeneic fecal material is contained in a liquid or solid dosage form.

21. 20. The composition of claim 19, wherein the allogeneic fecal material is contained in a capsule, tablet, or powder.

22. 22. The composition of any one of claims 1 to 21, wherein the subject is a mammal.

23. 23. The composition of claim 22, wherein the subject is a human.

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