IP Library Granted Patent US 12,414,969
Granted Patent B2
US 12,414,969 · App. 18/047,167 · Granted Sep 16, 2025

Methods and compositions for reducing clostridium difficile infection

Inventors: Eric Pamer (Guilford, CT); Charlie Buffie (New York, NY); Peter McKenney (New York, NY)
Assignee: Memorial Sloan-Kettering Cancer Center
A61K35/74A61K38/443A61K45/06C12N9/0006C12Q1/26C12Y101/01159
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Quick Facts
Patent No.
US 12,414,969
App. No.
18/047,167
Granted
Sep 16, 2025
Kind
B2
Abstract

The present invention relates to methods and compositions for reducing the risk and severity of C. difficile infection. It is based, at least in part, on the discovery that a restricted fraction of the gut microbiota, including the bacterium Clostridium scindens , contributes substantially to resistance against C. difficile infection. Without being bound by any particular theory, it is believed that this is achieved through the biosynthesis of secondary bile acids.

Claims (23)

1. A composition comprising two or more purified or isolated bacteria or spores thereof, the composition comprising:

a first purified or isolated bacteria or spores thereof comprising Clostridium scindens;

a second isolated or purified bacteria or spores thereof comprising Blautia hansenii or Barnesiella intestihominis ; and

a secondary bile acid selected from the group consisting of a deoxycholic acid, a lithocholic acid, and a combination thereof,

wherein the two or more purified or isolated bacteria or spores thereof are in a formulation for administration to a subject, wherein at least one of the two or more purified or isolated bacteria or spores thereof is capable of converting a primary bile acid to a secondary bile acid, and

wherein the composition comprises a capsule.

2. The composition of claim 1 , wherein the composition is suitable for being administered, to a subject in combination with an antibiotic, an immunotherapeutic agent, an herbal remedy, a probiotic, or combinations thereof.

3. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to inhibit proliferation of Clostridium difficile in the subject.

4. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to reduce the risk of Clostridium difficile infection and/or increase resistance to Clostridium difficile infection in a subject in need thereof.

5. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to reduce the severity of Clostridium difficile infection and/or decrease the amount of Clostridium Difficile toxin in a subject in need thereof.

6. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in combination with an enzyme that converts a bile acid to a secondary bile acid in an amount effective to decrease the severity of one or more symptoms of an intestinal disorder in a subject in need thereof.

7. The composition of claim 6 , wherein the symptoms are selected from the group consisting of frequency and/or volume of diarrhea; fever; abdominal cramping, pain, and/or tenderness; elevated level of white blood cells in the blood; loss of serum albumin; weight loss; appearance of pseudomembrane in the intestinal and/or rectal mucosa; and combinations thereof.

8. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to reduce one or more clinical symptoms of Clostridium difficile infection selected from the group consisting of frequency and/or volume of diarrhea; fever; abdominal cramping, pain, and/or tenderness; elevated level of white blood cells in the blood; loss of serum albumin; weight loss; appearance of pseudomembrane in the intestinal and/or rectal mucosa; and combinations thereof.

9. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to reduce the risk of developing a Clostridium difficile associated disease in a subject receiving antibiotic therapy.

10. The composition of claim 9 , wherein the Clostridium difficile -associated disease is Clostridium difficile colitis or pseudomembranous colitis.

11. The composition of claim 4 , wherein the Clostridium difficile infection comprises recurrent Clostridium difficile infection.

12. The composition of claim 1 , wherein the composition is suitable for being administered to a subject in an amount effective to convert a primary bile acid to a secondary bile acid in the subject.

13. The composition of claim 1 , wherein the composition comprises a combination of Clostridium scindens, Barnesiella intestihominis, Blautia hansenii , and Pseudoflavonifractor capillosus.

14. The composition of claim 1 , further comprising Pseudoflavonifractor capillosus.

15. The composition of claim 1 , wherein the composition is formulated for oral or rectal administration.

16. The composition of claim 1 , which further comprises a probiotic yeast.

17. The composition of claim 1 , further comprising an enzyme that converts a bile acid to a secondary bile acid.

18. The composition of claim 17 , wherein the enzyme that converts a bile acid to a secondary bile acid is a 7u-hydroxysteroid dehydrogenase enzyme.

Assignments (2)
RELEASE OF SECURITY INTEREST Recorded Sep 30, 2024
From: OAKTREE FUND ADMINISTRATION, LLC, AS ADMINISTRATIVE AGENT
To: SERES THERAPEUTICS, INC.
Reel/Frame 069082/0849 →
SECURITY INTEREST Recorded Apr 27, 2023
From: SERES THERAPEUTICS, INC.
To: OAKTREE FUND ADMINISTRATION, LLC
Reel/Frame 063485/0542 →
Continuity (4)
Division 16523414 · Jul 26, 2019
Continuation 15312610
Provisional Application 62000308 · May 19, 2014
Related Publication 20230190825A1 · Jun 22, 2023
References Cited (129)
US 7326551B2 · Maupin-Furlow et al. · 2008 [cited by applicant]
US 20040028689A1 · Borody · 2004 [cited by applicant]
US 20110280847A1 · Sorg et al. · 2011 [cited by applicant]
US 20140199281A1 · Henn et al. · 2014 [cited by applicant]
US 20170151291A1 · Henn · 2017 [cited by examiner]
JP 2001069990A · 2001 [cited by examiner]
WO WO2010062369A2 · 2010 [cited by examiner]
WO WO2012142605A1 · 2012 [cited by examiner]
WO WO2013053836A1 · 2013 [cited by applicant]
WO WO2013080561A1 · 2013 [cited by examiner]
WO WO2013171515A1 · 2013 [cited by applicant]
Kitahara et al. Int J Syst Evol Microbiol. 2000;50 Pt 3:971-978. [cited by examiner]
Wells et al. Clinica Chimica Acta 331.2003:127-134. [cited by examiner]
Ridlon, JM. Enzymology And Molecular Biology Of Bile Acid 7-alpha- And 7-beta-Dehydroxylation By The Intestinal Bacteria Clostridium scindens And Clostridium hylemonae. VCU Theses and Dissertations, 2008. [cited by examiner]
Definition of kit. http://oxforddictionaries.com/search?q=kit&view=uk retrieved Aug. 13, 2010. [cited by examiner]
U.S. Appl. No. 16/523,414 (11,471,495 B2) filed Jul. 26, 2019 (Oct. 18, 2022). [cited by applicant]
U.S. Appl. No. 15/312,610 (Abandoned) filed Nov. 18, 2016. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Jul. 15, 2022 Issue Fee Payment. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Jul. 11, 2022 Notice of Allowance. [cited by applicant]
U.S. Appl. No. 16/523,414, filed May 11, 2022 Response to Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Feb. 11, 2022 Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Dec. 10, 2021 Response to Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Jun. 11, 2021 Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Mar. 8, 2021 Response to Final Office Action with Request for Continued Examination (RCE). [cited by applicant]
U.S. Appl. No. 16/523,414, filed Nov. 6, 2020 Response to Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Sep. 9, 2020 Response to Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Jun. 9, 2020 Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 16/523,414, filed May 5, 2020 Response to Restriction Requirement. [cited by applicant]
U.S. Appl. No. 16/523,414, filed Feb. 5, 2020 Restriction Requirement. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Jul. 27, 2019 Abandonment. [cited by applicant]
U.S. Appl. No. 15/312,610, filed May 7, 2019 Advisory Action. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Apr. 26, 2019 Response after Final Office Action. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Feb. 26, 2019 Final Office Action. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Dec. 12, 2018 Response to Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Jul. 10, 2018 Non-Final Office Action. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Nov. 13, 2017 Response to Restriction Requirement. [cited by applicant]
U.S. Appl. No. 15/312,610, filed Sep. 13, 2017 Restriction Requirment. [cited by applicant]
U.S. Appl. No. 16/523,414, dated Jul. 15, 2022 Issue Fee Payment. [cited by applicant]
Abt et al., “Commensal Bacteria Calibrate the Activation Threshold of Innate Antiviral Immunity,” Immunity, 37(1): 158-170 (2012). [cited by applicant]
Atarashi et al., “Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota,” Nature, 500:232-236 (2013). [cited by applicant]
Bakken et al., “Treating Clostridium difficile infection with Fecal Microbiota Transplantation,” Clin Gastroenterol Hepatol., 9(12):1044-1049 (2011). [cited by applicant]
Barrasa et al., “Bile acids in the colon, from healthy to cytotoxic molecules,” Toxicology In Vitro 27:964-977 (2013). [cited by applicant]
Bartlett et al., “Antibiotic-Associated Pseudomembranous Colitis Due To Toxin- Producing Clostridia,” N. Engl. J. Med. 298(10):531-534 (1978). [cited by applicant]
Basler et al., “Tit-for-tat: Type VI secretion system counterattack during bacterial cell-cell interactions,” Cell, 152(4):884-894 (2013). [cited by applicant]
Basler et al., “Type VI secretion requires a dynamic contractile phage tail-like structure,” Nature, 483(7388): 182-186 (2013). [cited by applicant]
Bernstein et al., “Bile acids as carcinogens in human gastrointestinal cancers,” Mutation Res 589:47-65 (2005). [cited by applicant]
Brandl et al., “Vancomycin-resistant enterococci exploit antibiotic-induced innate immune deficit,”. Nature 455(7214): 804-807 (2008). [cited by applicant]
Britton et al., “Role of the Intestinal Microbiota in Resistance to Colonization by Clostridium difficile,” Gastroenterology 146:1547-1553 (2014). [cited by applicant]
Buffie et al., “Profound Alterations of Intestinal Microbiota following a Single Dose of Clindamycin Results in Sustained Susceptibility to [cited by applicant]
Buffie et al., “Microbiota-mediated colonization resistance against intestinal pathogens,” Nature Reviews Immunology 13(11):790-801 (2013). [cited by applicant]
Buffie et al., “Precision microbiome reconstitution restores bile acid mediated resistance to [cited by applicant]
Caporaso et al., “QIIME allows analysis of high-throughput community sequencing data,” Nat Methods. 7(5):335-336 (2010). [cited by applicant]
Caporaso et al., “Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms,” The ISME Journal 6:1621-1624 (2012). [cited by applicant]
Carlier et al., “Proposal to unify [cited by applicant]
Chang et al., “Decreased Diversity of the Fecal Microbiome in Recurrent [cited by applicant]
Chen et al., “A Mouse Model of [cited by applicant]
Chen et al., “Overview of [cited by applicant]
Chung et al., “Gut Immune Maturation Depends on Colonization with a Host-Specific Microbiota,” Cell 149(7): 1578-1593 (2012). [cited by applicant]
Cohen, Statistical Powerr Analysis for the Behavioral Sciences, Second Edition Routledge, Hillsdale, NJ, 1988). [cited by applicant]
Collins et al., “The Phylogeny of the Genus Clostridium: Proposal of Five New Genera and Eleven New Species Combinations,” Int J Syst Bacteriol 44(4):812-826 (1994). [cited by applicant]
Cruz et al. Antimicrob. Agents Chemother. Jan. 2000 vol. 44 No. 1 143-149. [cited by applicant]
De Aguiar Vallim et al., “Pleiotropic Roles of Bile Acids in Metabolism,” Cell Metab. 17(5):657-669 (2013). [cited by applicant]
Dethlefsen et al., “Incomplete recovery and individualized responses of the human distal gut microbiota to repeated antibiotic perturbation,” PNAS 108(Suppl. 1):4554-4561 (2011). [cited by applicant]
Diehl et al., “Microbiota Restricts Trafficking of Bacteria to Mesenteric Lymph Nodes by CX(3)CRI(hi) Cells,” Nature 494(7435):116-120 (2013). [cited by applicant]
Duan et al., “Microbial colonization drives expansion of IL-1 receptor 1 expressing, IL-17 producing gamma/delta T cells,” Cell Host Microbe, 7(2): 140-150 (2010). [cited by applicant]
Edgar et al., “UCHIME improves sensitivity and speed of chimera detection,” Bioinformatics 27(16):2194-2200 (2011). [cited by applicant]
Farache et al., “Luminal Bacteria Recruit CD103(+) Dendritic Cells into the Intestinal Epithelium to Sample Bacterial Antigens for Presentation,” Immunity, 38(3):581-595 (2013). [cited by applicant]
Ferreira et al., “The Intestinal Microbiota Plays a Role in Salmonella-Induced Colitis Independent of Pathogen Colonization,” PLoS One 6(5):e20338 (2011). [cited by applicant]
Giel et al., “Metabolism of Bile Salts in Mice Influences Spore Germination in Clostridium difficile,” PLoS ONE, 5(1):e8740 (2010). [cited by applicant]
“Gut” definition. Merriam Webster Dictionary. https://www.merriam-webster.com/dictionary/gut retrieved Feb. 20, 2019/. [cited by applicant]
Hall, “Building Phylogenetic Trees from Molecular Data with MEGA,” Mol. Biol. Evol. 30(5):1229-1235 (2013). [cited by applicant]
Hamilton et al., “High-throughput DNA sequence analysis reveals stable engraftment of gut microbiota following transplantation of previously frozen fecal bacteria,” Gut Microbes 4(2): 125-135 (2013). [cited by applicant]
Hand et al., “Acute Gastrointestinal Infection Induces Long-Lived Microbiota-Specific T Cell Responses,” Science, 337(6101): 1553-1556 (2012). [cited by applicant]
Heeg et al., “ [cited by applicant]
Hill et al., “Commensal bacteria-derived signals regulate basophil hematopoiesis and allergic inflammation,” Nat Med., 18(4):538-546 (2012). [cited by applicant]
Huse et al., “Exploring Microbial Diversity and Taxonomy Using SSU rRNA Hypervariable Tag Sequencing,” PLoS Genet 4(11):e1000255 (2008). [cited by applicant]
International Search Report mailed Sep. 8, 2015 in International Application No. PCT/US15/31627. [cited by applicant]
Ivanov et al., “Induction of intestinal Th17 cells by segmented filamentous bacteria,” Cell, 139(3):485-498 (2009). [cited by applicant]
Kang et al., “ [cited by applicant]
Kinnebrew et al., “Early [cited by applicant]
Koeth et al., “Intestinal microbiota metabolism of L-carnitine, a nutrient in red meat, promotes atherosclerosis,” Nat Med. 19(5):576-585 (2013). [cited by applicant]
Krishna et al., “Risk Factors, preemptive therapy, and antiperistaltic agents for Clostridium difficile infection in cancer patients,” Transplant Infect Dis., 15:493-501 (2013). [cited by applicant]
Kyne et al., “Health Care Costs and Mortality Associated with Nosocomial Diarrhea Due to [cited by applicant]
Langille et al., “Predictive functional profiling of microbial communities using 16S rRNA marker gene sequences,” Nat Biotechnol 31(9):814-821 (2013). [cited by applicant]
Lathrop et al., “Peripheral education of the immune system by colonic commensal microbiota,” Nature, 478(7368):250-254 (2012). [cited by applicant]
Lawley et al., “Targeted Restoration of the Intestinal Microbiota with a Simple, Defined Bacteriotherapy Resolves Relapsing [cited by applicant]
Liu et al., “Reclassification of [cited by applicant]
Louie et al., “Tolevamer, a Novel Nonantibiotic Polymer, Compared with Vancomycin in the Treatment of Mild to Moderately Severe [cited by applicant]
Lozupone et al., “UniFrac: a New Phylogenetic Method for Comparing Microbial Communities,” Appl Environ Microbiol 71(12):8228-8235 (2005). [cited by applicant]
Macpherson et al., “Induction of Protective IgA by Intestinal Dendritic Cells Carrying Commensal Bacteria,” Science, 303:1662-1665 (2004). [cited by applicant]
Manges et al., “Comparative Metagenomic Study of Alterations to the Intestinal Microbiota and Risk of Nosocomial [cited by applicant]
Marcus et al. Gut, 1988, 29, 522-533. [cited by applicant]
Marsh et al., “Association of Relapse of [cited by applicant]
Olszak et al., “Microbial Exposure During Early Life has Persistent Effects on Natural Killer T Cell Function,” Science, 336(6080):489-493 (2012). [cited by applicant]
Ott et al., “Quantification of Intestinal Bacterial Populations by Real-Time PCR with a Universal Primer Set and Minor Groove Binder Probes: a Global Approach to the Enteric Flora,” Journal of Clinical Microbiology, 42(… [cited by applicant]
Out et al., “Bile acid sequestrants: more than simple resins,” Curr Opin Lipidol 23:43-55 (2012). [cited by applicant]
Pamer, “Fecal microbiota transplantation: effectiveness, complexities, and lingering concerns,” Mucosal Immunol 7(2):210-214 (2014). [cited by applicant]
Partial Supplementary European Search Report dated Jan. 4, 2018 in Application No. 15796000.6. [cited by applicant]
Petrof et al., “Stool substitute transplant therapy for the eradication of Clostridium difficile infection: 'RePOOPulating' the gut,” Microbiome 1:3, pp. 1-12 (2013). [cited by applicant]
Rakoff-Nahoum et al., “Recognition of Commensal Microflora by Toll-Like Receptors Is Required for Intestinal Homeostasis,” Cell, 118:229-241 (2004). [cited by applicant]
Rasti et al. Journal of Food Agriculture and Environment 11(2):127-131, Apr. 2013. [cited by applicant]
Rea et al., “Effect of broad- and narrow-spectrum antimicrobials on Clostridium difficile and microbial diversity in a model of the distal colon,” PNAS 108(Suppl. 1):4639-4644 (2011). [cited by applicant]
Rea et al., “Thuricin CD, a posttranslationally modified bacteriocin with a narrow spectrum of activity against [cited by applicant]
Reeves et al., “The interplay between microbiome dynamics and pathogen dynamics in a murine model of [cited by applicant]
Ridlon et al., “Bile salt biotransformations by human intestinal bacteria,” J Lipid Res 47:241-259 (2006). [cited by applicant]
Ridlon et al., “ [cited by applicant]
Ridlon et al., “Identification and characterization of two bile-acid coenzyme A transferases from [cited by applicant]
Rupnik et al., “Clostridium difficile infection: new developments in epidemiology and pathogenesis,” Nat Rev Microbiol 7:526-536 (2009). [cited by applicant]
Schloss et al., “Introducing mothur: Open-Source, Platform-Independent, Community-Supported Software for Describing and Comparing Microbial Communities,” Appl Environ Microbiol 75(23):7537-7541 (2009). [cited by applicant]
Sheneman et al., “Clearcut: a fast implementation of relaxed neighbor joining,” Bioinformatics 22(22):2823-2824 (2006). [cited by applicant]
Sorg et al., “Bile Salts and Glycine as Cogerminants for [cited by applicant]
Sorg et al., “Chenodeoxycholate is an Inhibitor of [cited by applicant]
Stein et al., “Ecological Modeling from Time-Series Inference: Insight into Dynamics and Stability of Intestinal Microbiota,” PLoS Comput Biol 9(12):e1003388 (2013). [cited by applicant]
Surawicz et al., “Treatment of refractory and recurrent [cited by applicant]
Taur et al., “Intestinal Domination and the Risk of Bacteremia in Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation,” Clin Infect Dis 55(7):905-914 (2012). [cited by applicant]
The Human Microbiome Project Consortium: “Structure, Function and Diversity of the Healthy Human Microbiome,” Nature 486(7402):207-214 (2013). [cited by applicant]
Theriot et al., “Antibiotic-induced shifts in the mouse gut microbiome and metabolome increase susceptibility to [cited by applicant]
Turnbaugh et al., “A core gut microbiome in obese and lean twins,” Nature 457(7228): 480-484 (2009). [cited by applicant]
Ubeda et al., “Intestinal Microbiota Containing [cited by applicant]
Ubeda et al., “Vancomycin-resistant [cited by applicant]
van Nood et al., “Duodenal Infusion of Donor Feces for Recurrent [cited by applicant]
Vogt et al. Anaerobe 34 (2015)106-115. [cited by applicant]
Weingarden et al., “Microbiota transplantation restores normal fecal bile acid composition in recurrent [cited by applicant]
Wells et al., “Identification and Characterization of a Bile Acid 7β-Dehydroxylation Operon in [cited by applicant]
Wingender et al., “Intestinal Microbes Affect Phenotypes and Functions of Invariant Natural Killer T Cells in Mice,” Gastroenterology, 143(2):418-428 (2012). [cited by applicant]
Yutin et al., “A genomic update on clostridial phylogeny: Gram-negative spore formers and other misplaced clostridia,” Environ Microbiol. 15(10):2631-2641 (2013). [cited by applicant]
Zar et al., “A Comparison of Vancomycin and Metronidazole for the Treatment of [cited by applicant]
Zhao et al., “RAPSearch2: a fast and memory-efficient protein similarity search tool for next-generation sequencing data,” Bioinformatics 28(1): 125-126 (2012). [cited by applicant]
Zilberberg et al., “Increase in Adult [cited by applicant]