Microbiota restoration therapy (mrt), compositions and methods of manufacture

Abstract

Microbiota restoration therapy compositions and methods for manufacturing, processing, and/or delivering microbiota restoration therapy compositions are disclosed. An example method for manufacturing a microbiota restoration therapy composition may include collecting a human fecal sample and adding a diluent to the human fecal sample to form a diluted sample. The diluent may include a cryoprotectant. The method may also include mixing the diluted sample with a mixing apparatus and filtering the diluted sample. Filtering may form a filtrate. The method may also include transferring the filtrate to a sample bag and sealing the sample bag.

What is claimed is: 
     
         1 . A microbiota restoration therapy composition, comprising:
 a mixture of an effective amount of fecal microbiota and an effective amount of a cryoprotectant;   wherein the cryoprotectant includes dextrose, betaine, glycine, sucrose, polyvinyl alcohol, Pluronic F-127, Mannitol, tween 80, ethylene glycol, 1,3-propanediol, hydroxypropyl cellulose, a mixture of polyethylene glycol and glycerol, propylene glycol, or combinations thereof.   
     
     
         2 . The microbiota restoration therapy composition of  claim 1 , wherein the fecal microbiota is derived from one or more human stool samples and wherein the viability of the microbiota is confirmed by culturing the microbiota on a  Bacteroides  Bile Esculin Agar (BBE) plate, a Center for Disease Control (CDC) plate, or both. 
     
     
         3 . The microbiota restoration therapy composition of  claim 2 , wherein the viability of the microbiota is confirmed on a BBE plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −5 . 
     
     
         4 . The microbiota restoration therapy composition of  claim 2 , wherein the viability of the microbiota is confirmed on a CDC plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −6 . 
     
     
         5 . The microbiota restoration therapy composition of  claim 1 , wherein the fecal microbiota contains at least 10 7  microbes/ml. 
     
     
         6 . The microbiota restoration therapy composition of  claim 1 , wherein the fecal microbiota is derived from human stool samples that are obtained from donors that have been pre-screened and post-screened at an interval of about 30 days to about 100 days to assure a healthy sample was collected in the interval therebetween. 
     
     
         7 . The microbiota restoration therapy composition of  claim 1 , wherein adding a diluent to the fecal microbiota to form a diluted sample includes adding a mixture of saline and the cryoprotectant to the fecal microbiota. 
     
     
         8 . The microbiota restoration therapy composition of  claim 1 , further comprising storing the microbiota restoration therapy composition in a temperature specific storing device at a temperature of about −20° C. to about −80° C. 
     
     
         9 . A method for producing a microbiota restoration therapy composition from a human stool sample, the method comprising:
 collecting a stool sample from a human donor;   adding an amount of saline to the stool sample;   adding an effective amount of a cryoprotectant;   wherein the cryoprotectant includes dextrose, betaine, glycine, sucrose, polyvinyl alcohol, Pluronic F-127, Mannitol, tween 80, ethylene glycol, 1,3-propanediol, hydroxypropyl cellulose, a mixture of polyethylene glycol and glycerol, propylene glycol, glycerol, or combinations thereof;   mixing the stool sample, saline, and the cryoprotectant together to make a mixed composition;   filtering the mixed composition and collecting the filtrate;   removing a portion of the filtrate for testing and freezing the remainder of the filtrate;   storing the frozen filtrate under quarantine;   testing the portion of the filtrate to measure the viability and quality of microbiota within the filtrate;   confirming the viability and quality of the filtrate; and   releasing the filtrate from quarantine to define a microbiota restoration therapy composition.   
     
     
         10 . The method of  claim 9 , wherein the viability of the microbiota within the filtrate is confirmed by culturing the microbiota on a  Bacteroides  Bile Esculin Agar (BBE) plate, a Center for Disease Control (CDC) plate, or both. 
     
     
         11 . The method of  claim 10 , wherein the viability of the microbiota within the filtrate is confirmed on a BBE plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −5 . 
     
     
         12 . The method of  claim 10 , wherein the viability of the microbiota within the filtrate is confirmed on a CDC plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −6 . 
     
     
         13 . The method of  claim 9 , wherein the filtrate contains at least 10 7  microbes/ml. 
     
     
         14 . The method of  claim 9 , wherein the human donor is pre-screened and post-screened at an interval of about 30 days to about 100 days to assure a healthy sample was collected in the interval therebetween. 
     
     
         15 . A method for assuring the quality of a human stool sample to be processed into a microbiota restoration therapy composition, the method comprising:
 identifying a human stool donor;   conducting a pre-donation screening of the donor, comprising a health history questionnaire and at least one blood test;   collecting a human stool sample from the donor;   processing the stool sample from the donor to form one or more microbiota restoration therapy compositions;   wherein processing the stool sample includes mixing an effective amount of a cryoprotectant;   wherein the cryoprotectant includes dextrose, betaine, glycine, sucrose, polyvinyl alcohol, Pluronic F-127, Mannitol, tween 80, ethylene glycol, 1,3-propanediol, hydroxypropyl cellulose, a mixture of polyethylene glycol and glycerol, propylene glycol, glycerol, or combinations thereof;   conducting a post-donation screening of the donor at an interval of about 30 days to about 100 days, comprising a health history questionnaire and at least one blood test;   holding in quarantine the one or more microbiota restoration therapy compositions processed from the stool sample collected during the interval between pre-donation screening and post-donation screening;   confirming the quality of the one or more microbiota restoration therapy compositions from both pre- and post-screening results; and   releasing the one or more microbiota restoration therapy compositions from quarantine.   
     
     
         16 . The method of  claim 15 , further comprising conducting at least one test on the human stool sample for the presence of a constituent selected from the group comprising  C. difficile ; Norovirus; Adenovirus; Enteric Pathogens;  Giardia  antigen;  Cryptosporidium  antigen; Acid-fast staining (Clyslospora, Isospora); ova and parasites; Vancomycin-resistant enterococci (VRE); Methicillin-resistant  Staphylococcus aureus  (MRSA); and combinations thereof. 
     
     
         17 . The method of  claim 16 , wherein the blood test includes at least one test for a constituent selected from the group comprising: HIV; Hepatitis A; Hepatitis B; Hepatitis C; RPR and combinations thereof. 
     
     
         18 . The method of  claim 15 , further comprising confirming the viability of the one or more microbiota restoration therapy compositions by culturing the one or more microbiota restoration therapy compositions on a  Bacteroides  Bile Esculin Agar (BBE) plate, a Center for Disease Control (CDC) plate, or both. 
     
     
         19 . The method of  claim 15 , wherein the viability of the one or more microbiota restoration therapy compositions is confirmed on a BBE plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −5 . 
     
     
         20 . The method of  claim 15 , wherein the viability of the one or more microbiota restoration therapy compositions is confirmed on a CDC plate by a colony forming unit (CFU) count of about 30 CFU to about 300 CFU at a serial dilution of 10 −6 .