MANAGEMENT OF MICROBIAL DYSBIOSIS

Abstract

Methods for using temocillin to control microbial dysbiosis arising, in certain cases, from enterobacterial overgrowth, such as invasive adherent E. coli (IAEC) overgrowth, are provided herein. Methods for using temocillin to treat intestinal disorders and their symptoms caused by microbial dysbiosis, as well as other disorders, are also considered herein.

Description

MANAGEMENT OF MICROBIAL DYSBIOSIS RELATED APPLICATIONS This application claims the benefit of priority of U.S. Provisional Application No. 62 / 952,874, filed on December 23, 2019, and U.S. Provisional Application No. 62 / 958,979, filed on January 9, 2020, the contents of which are incorporated herein by reference in their entirety. BACKGROUND OF THE INVENTION The human gastrointestinal (GI) tract harbors a complex and dynamic population of over 100 trillion microorganisms. The collection of bacteria, archaea, and eukaryotes that colonize the GI tract is known as the gut microbiota. The gut microbiota establishes an intricate and mutually beneficial relationship with the host, i.e., the human carrier, and provides many benefits through multiple physiological functions, such as strengthening intestinal integrity, shaping the intestinal epithelium, harvesting energy, protecting against pathogens, and regulating host immunity. With the advent of modern technology, the involvement of the gut microbiota in a variety of intestinal and extraintestinal conditions has become readily apparent. See, e.g., Schroeder et al., Nat. Med. 22, 1079–1089 and Chang et al., Best Pract. Res. Clin. Gastroenterol. 30, 3–15. Microbial dysbiosis (also known as dysbacteriosis) is a term that refers to a disruption or imbalance of the normal microbiome composition in the GI tract. This imbalance may result from the gain or loss of one or more microbes with functions detrimental to the host or from the loss of one or more microbes with functions beneficial to the host. In either case, if left untreated, dysbiosis can contribute to a plethora of diseases ranging from intestinal disorders (e.g., irritable bowel syndrome, Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis) to neurodevelopmental disorders. See, for example, Guinane et al., 2013, Therap Adv Gastroenterol.6, 295-308. In patients with intestinal disorders resulting from microbial dysbiosis, higher populations of proteobacteria, such as Enterobacteriaceae, have been observed. See, for example, Zeng et al., Mucosal Immunol. 2017 Jan; 10(1): 18-26 and Serino J Mol Biol. 2018 Mar 2; 430(5):581-590. In particular, recent evidence suggests that disruption of the microbial community favors the emergence of invasive adherent E. coti (IAEC), which can increase the incidence and severity of intestinal inflammation in the context of Crohn's disease and ulcerative colitis (see BioMed Research International, vol. 2014, article ID 567929, 16 pages), in addition to playing a role in colorectal cancer and intestinal disease in animals. See, for example, Rahmouni et al., Gut Pathogens 2018, 10:23. Two AIEC phylotypes (B2 and D) have been described as more prevalent in patients with IBD (see PLoS One, 2019, 14(4): e0216165 and Gut, 2007, 56:669-675). Infectious inflammatory cells (IICs) affect host cell processes such as protein synthesis, signal transduction, cell division, ion secretion, transcription, cytoskeleton function, and mitochondrial function. They are particularly difficult to treat and can adhere to the intestinal mucosa by binding to carcinoembryonic antigen-related cell adhesion molecules 6 (CEACAM6), invade intestinal epithelial cells using host cell actin microfilaments and microtubules, replicate intracellularly, translocate across the human intestinal barrier, and move into deeper tissues. IICs can also survive within macrophages, stimulate TNF-α production, and promote a granulomatous inflammatory response. Due to their overabundance during microbial dysbiosis, their invasiveness and lack of sensitivity to various antibiotic treatments (see Dogan et al, Inflamm Bowel Dis. 2013 Jan;19(l): 141-50.), and their subsequent implications associated with intestinal infections and other disorders, there is a need to control AIEC overgrowth. SUMMARY OF THE INVENTION Temocillin has now been found to be an effective modulator of invasive adherent E. coül (IAEC) overgrowth and, in certain cases, performs better than the current standard treatment, ciprofloxacin. See, for example, Figure 1, which shows that temocillin was effective in reducing the IAEC burden in the murine GI tract, and Table 1, which provides data showing lower MIC values ​​for temocillin (MICs = 4 or 8 pg / mL) than for ciprofloxacin (MICs > 8 pg / mL) against IAEC strains in vitro. In another aspect, temocillin was found to have little to no effect on normal GI flora; for example, it was determined to be microbiome-preserving for at least Firmicutes, Actinobacteria, and Bacteroidetes. This contrasts with ciprofloxacin, rifaximin, and metronidazole, each of which had broad and potent antibacterial activity against Firmicutes, Actinobacteria, and Bacteroidetes. See, for example, Table 2. This is another added benefit of temocillin treatment in that there is little concern about a failure to restore normal, healthy flora. In another aspect, temocillin was found to be more effective in reducing the probability of recurrence of AIEC overgrowth after treatment. See, for example, Figure 1, where AIEC bacterial growth remained low for at least 7 days after treatment with temocillin. However, ciprofloxacin showed a rebound effect. See, for example, Figure 1. Therefore, uses of temocillin, or one of its pharmaceutically acceptable salts, for the treatment of microbial dysbiosis are provided herein. Also provided herein are uses of temocillin, or a pharmaceutically acceptable salt thereof, for treating an intestinal disorder in a subject experiencing microbial dysbiosis. Methods are also provided for modulating enterobacterial overgrowth (e.g., AIEC overgrowth) in a subject experiencing microbial dysbiosis or intestinal disorder, or both. Methods for treating one or more symptoms associated with microbial dysbiosis are also provided herein. In addition, methods for treating one or more symptoms of a disorder caused by microbial dysbiosis are provided herein. In one respect, the aforementioned microbial dysbiosis is caused by an overgrowth of enterobacteria (e.g., AIEC overgrowth). Other uses of temocillin and other aspects are described in detail below. BRIEF DESCRIPTION OF THE FIGURES Figure 1 represents the bacterial load in feces versus days obtained from the murine model study of AIEC GI decolonization with temocillin and ciprofloxacin. Figure 2A represents the amount of OTU (indication of richness) obtained from post-treatment measurement of microbiome health for temocillin and ciprofloxacin in mice. Figure 2B represents the Shannon entropy index (indication of diversity) obtained from post-treatment measurement of microbiome health for temocillin and ciprofloxacin in mice. DETAILED DESCRIPTION OF THE INVENTION A. Definitions As used herein, temocillin refers to the compound having the formula: hb? Qan / zznz / q / YILI EITHER Unless otherwise specified, temocillin includes the stereoisomer described above, as well as all other possible stereoisomers and mixtures of stereoisomers. However, in one respect, the uses described herein refer to stereoisomer 25, 5 / ?, 65 represented with a stereochemical enrichment or molar excess of at least 60%, at least 65%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, or at least 99%. Enterobacteriaceae include, but are not limited to, Escherichia co / i (E. co / i), Klebsiella, Salmonella, Shigella, and Yersinia pestis. As used herein, AIEC refers to invasive adherent E. coH. In one respect, AIEC can adhere to and invade cultured intestinal epithelial cells, survive, and multiply within macrophages without expressing virulence factors or genes associated with diarrheagenic E. cu / / and uropathogenic E. coH (UPEC) pathotypes. In one respect, the characteristics of AIEC include, but are not limited to, (i) the ability to adhere to and invade intestinal epithelial cells, (ii) the ability to survive and replicate expansively within macrophages without triggering host cell death, (iii) eliciting significantly elevated expression of TNF-alpha and anthokines, and (iv) the lack of known invasive determinants of the traditional Type III virulence / invasion-related machinery. See, e.g., J Crohns Colitis. 2015 May;9(5):410-20; Lab Invest. 2012 Mar;92(3):411-9; and Lab Invest. 2012 Mar;92(3):411-9.AIEC characterization can be achieved by following routine procedures known to an expert in the technique or procedures (e.g., adhesion assay, invasion assay, and virulence genotyping assay) described in Rehmouni et al., Gut Pathogens, 2018, 10:23. Intestinal disorders refer to conditions or diseases that occur in or affect the intestine. Intestinal disorders include, but are not limited to, irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis. The terms subject and patient can be used interchangeably and mean a mammal in need of treatment, for example, companion animals (e.g., dogs, cats, and the like), farm animals (e.g., cows, pigs, horses, sheep, goats, and the like), and laboratory animals (e.g., rats, mice, guinea pigs, and the like). Typically, the subject is a human being in need of treatment. As used herein, the term "treat" or "treatment" refers to achieving the desired pharmacological and / or physiological effect. The effect may be therapeutic, which includes achieving, partially or substantially, one or more of the following results: partially or totally reducing the extent of the disease, disorder, or syndrome; relieving or improving a symptom or clinical indicator associated with the disorder; or delaying, inhibiting, or decreasing the likelihood of progression of the disease, disorder, or syndrome. In one respect, treatment may also continue after symptoms have resolved, for example, to delay their recurrence. "Treat" or "treatment" also refers to inhibiting and / or delaying the rebound of the infection or the symptoms of the infection. As used herein, the term modular means to reduce, inhibit, control, improve, or maintain. The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, timing of administration, excretion rate, drug combinations, the treating physician's judgment, and the severity of the particular disease being treated. However, in one respect, the temocillin described in these methods is administered in an effective or therapeutically effective amount. This refers to an amount of temocillin that will elicit a biological or medical response in a subject, for example, a dose of between 0.01 and 100 mg / kg of body weight / day. In certain aspects, the temocillin described herein can be administered orally, topically, rectally, nasally, buccally, or via an implanted reservoir. The term parenteral, as used herein, includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the temocillin described herein is administered orally or rectally. In one aspect, the temocillin described herein is administered orally. The temocillin described in these methods can be formulated to provide rapid, sustained, or delayed release of temocillin after administration to the patient using procedures known in the art. In one respect, the temocillin described in these methods is formulated as a delayed-release oral composition. The temocillin salts described herein refer to pharmaceutically acceptable, non-toxic salts. Pharmaceutically acceptable salt forms include pharmaceutically acceptable acid / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts include, for example, salts of inorganic acids (such as hydrochloric, hydrobromic, phosphoric, nitric, and sulfuric acids) and organic acids (such as acetic, benzenesulfonic, benzoic, methanesulfonic, and toluenesulfonic acids). Suitable pharmaceutically acceptable basic salts include, for example, ammonium salts, alkali metal salts (such as sodium and potassium salts), and alkaline earth metal salts (such as magnesium and calcium salts). In one respect, the pharmaceutically acceptable salt is a sodium salt. The term overgrowth refers to increases in the abundance of one or more bacteria that go beyond what is expected in healthy humans consuming a typical Western diet. B. Uses In a first embodiment, a method is provided herein for modulating (e.g., reducing) microbial dysbiosis in a subject in need, comprising administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In one aspect, the microbial dysbiosis in the first embodiment is caused by enterobacterial overgrowth, for example, AIEC overgrowth. In a second embodiment, a method is provided herein for treating an intestinal disorder in a subject experiencing microbial dysbiosis, comprising administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In one aspect, the microbial dysbiosis of the second embodiment is caused by an overgrowth of enterobacteria, for example, an overgrowth of AIEC. In a third embodiment, a method is provided herein for reducing one or more of the following symptoms in a subject experiencing microbial dysbiosis: inflammation, diarrhea, fever, fatigue, abdominal pain and cramps, weight loss, and anemia. This method comprises administering to the subject an effective amount of temocillin or a pharmaceutically acceptable salt thereof. In one aspect, the microbial dysbiosis of the third embodiment is caused by an overgrowth of enterobacteria, for example, an overgrowth of AIEC. In a fourth embodiment, a method is provided herein for reducing one or more of the following symptoms: inflammation, diarrhea, fever, fatigue, abdominal pain and cramps, weight loss, and anemia in a subject with an intestinal disorder who also experiences microbial dysbiosis. This method comprises administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In one aspect, the microbial dysbiosis of the fourth embodiment is caused by an overgrowth of enterobacteria, for example, an overgrowth of AIEC. In a fifth embodiment, a method is provided herein for treating a bowel disorder in a subject experiencing invasive adherent E. coli (IAEC) overgrowth in the intestine, comprising administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In one aspect, the bowel disorder of the fifth embodiment is selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome associated with diarrhea (d-IBS), irritable bowel syndrome with constipation (cIBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis. In one aspect, the bowel disorder of the fifth embodiment is Crohn's disease. In a sixth embodiment, a method is provided herein for modulating (e.g., reducing) the overgrowth of invasive adherent E. coli (IAEC) in a subject in need, comprising the step of administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In one aspect, the IAEC overgrowth in the sixth embodiment is in the subject's intestine. In one aspect, the subject of the sixth embodiment has an intestinal disorder. In one aspect, the subject of the sixth embodiment has an intestinal disorder and has IAEC overgrowth in the intestine.In one aspect, the subject of the sixth embodiment has a bowel disorder selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis. In one aspect, the subject of the sixth embodiment has a bowel disorder selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis and has intestinal invasive gastrointestinal endocrine disruption (IGICD). In one aspect, the subject of the sixth embodiment has Crohn's disease.In one aspect, the subject of the sixth embodiment has Crohn's disease and intestinal invasive gastrointestinal encephalopathy (IIG). In another aspect, the subject of the sixth embodiment has a bowel disorder selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome associated with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, pouchitis, and diverticulitis.In one aspect, the subject of the sixth realization has an intestinal disorder that is selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome associated with diarrhea (d-IBS), irritable bowel syndrome with constipation (cIBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, pouchitis, and diverticulitis, and has an AIEC overgrowth of the intestine. In a seventh embodiment, a method is provided herein for reducing intestinal inflammation caused by invasive adherent E. coli (IAEC) overgrowth in a subject in need (e.g., having Crohn's disease), comprising the step of administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof. In an eighth embodiment, a method is provided herein for reducing one or more of the symptoms of diarrhea, fever, fatigue, abdominal pain and cramps, weight loss, and anemia in a subject with Crohn's disease, comprising administering to the subject orally an effective amount of temocillin or a pharmaceutically acceptable salt thereof. In one aspect, the subject in the eighth embodiment is experiencing an overgrowth of invasive adherent E. coli (IAEC) of the intestine. In a ninth embodiment, a subject of the present methods is administered a pharmaceutically acceptable salt of temocillin. In a tenth embodiment, temocillin or a pharmaceutically acceptable salt thereof described in the present methods is administered orally. In an eleventh embodiment, the methods described herein inhibit the overgrowth of Enterobacteriaceae (e.g., AIEC overgrowth) without inhibiting the growth of Firmicutes or Actinobacteria bacteria, or both. In a twelfth embodiment, administration of temocillin reduces the likelihood of recurrence of Enterobacteriaceae overgrowth (e.g., AIEC overgrowth) for a period of at least 7 days after treatment. The invention can be better understood with reference to the following illustrative examples, which are intended to exemplify non-limiting embodiments of the invention. EXAMPLES General methods Temocillin disodium salt was obtained from BOC Sciences (45-16 Ramsey Road, Shirley, NY 11967, USA) and Bellen Chemistry (1 Caída, 3rd Street, MaoHua Garden, Beijing, China 101300) and represented a mixture of two diastereomers (R, S) in a (60, 40) ratio. Ciprofloxacin hydrochloride was obtained from USP-US (Rockville, MD, USA). Temocillin stocks for all in vivo and in vitro tests were dissolved in 0.9% sodium chloride, and water was used as the solvent for ciprofloxacin. Invasive adherent E. coli (AIEC) strains were acquired from the Thomas S. Whittam STEC Center (194 Food Safety and Toxicology, Michigan State University, MIC 48824). Example 1 Temocillin activity versus AIEC The antibacterial activity of temocillin was tested against AIEC and compared with ciprofloxacin. Briefly, temocillin or ciprofloxacin stock solutions were diluted two-fold and incubated with the appropriate strain in sterile 96-well polystyrene microplates at 35 °C. The plates were visually read after 16–18 h of incubation to identify minimum inhibitory concentration (MIC) values. As shown in Table 1, temocillin is effective at doses of 4–8 pg / mL against at least 50% of the tested clinical AIEC isolates from Crohn's disease and maintains antibacterial activity against ciprofloxacin-resistant isolates. Table 1 Crohn's AIEC isolates No. 14 No. 32 No. 40 No. 51 No. 35 No. 49 Compound MIC, pg / mL Temocillin 16 4 8 8 16 4 Ciprofloxacin >8 >8 >8 >8 >8 8 Example 2 Murine model study of decolonization with temocillin and ciprofloxacinMurine model studies of GI AIEC decolonization were conducted using a previously published murine model as a basis and performed in accordance with animal ethics guidelines at an external contract research organization. Healthy C57BL / 6J mice were first administered streptomycin via oral gavage. They were then administered an AIEC strain (Phylotype D) at approximately 1–5 x 10⁶ cfu per mouse via oral gavage, and fecal sediments were enumerated to determine the AIEC load. After stable colonization (typically 48 hours after AIEC administration), the mice were divided into three groups of four animals each. One group received ciprofloxacin, while another received temocillin via oral gavage (both at 30 mg / kg). The third group served as the control and received an equivalent volume of sterile saline via oral gavage.Fecal sediments were collected at various time points as indicated on the X-axis of Figure 1, and the AIEC load was quantified by dilution and plating. Two successive fecal sediments were collected from each mouse per group to ensure sufficient data potency. Temocillin was effective in reducing the AIEC burden in the murine GI tract and performed as well as the current reference antibiotic, ciprofloxacin (Figure 1). Of particular importance, temocillin was effective in maintaining a low AIEC burden for days after treatment discontinuation, whereas mice treated with ciprofloxacin began to show a rebound effect after day 7 post-treatment discontinuation (Figure 1). This supports temocillin's ability to reduce the likelihood of recurrent AIEC overgrowth after treatment. Example 3 Microbiome conservative nature of temocillin Representatives of anaerobic commensals of the human gut belonging to the three main phyla, Firmicutes, Actinobacteria, and Bacteroidetes, were analyzed in standard agar dilution antibacterial assays performed under anaerobic conditions. The work was carried out externally at a microbiology contract research organization, and the results are presented in Table 2. Firmicutes and Actinobacteria are part of the healthy human gut, and their levels are compromised in patients with Crohn's disease. As shown, temocillin has little effect (MIC >256 pg / mL) on certain Firmicutes, Actinobacteria, and Bacteroidetes. Table 2 MIC, pg / mL Firmicutes Temocillin Ciprofloxacin Rifaximin Metronidazole MMX1722 LadobaciHus case / >256 1 0.25 >64 MMX 6660 Streptococcus intermedius >256 1 0.5 >64 A TCC23272 LactobaciHus reuteri >256 32 0.06 >64 MMX 9797 Ciostridium innocuum >256 4 >32 1 Actinobacteria A TCC 15702 Bifidobacterium infantis >256 4 0.125 8 A TCC25986 Collinsella aerofaciens >256 0.5 <0.03 1 MMX 10069 Bifidobacterium adoiescentis >256 32 0.5 8 Bacteroidetes A TCC 12290 Bacteroides thetaiotaomicron >256 1 <0.03 0.25 MMX 9711 Bacteroides saiyersiae >256 16 0.5 2 MMX3478 Bacteroides fragilis 256 16 0.25 8 Proteobacteria Crohn's patient no. ° 32 AIEC 4 >8 nd nd Crohn's patient no. ° 40 AIEC 8 >8 nd nd Crohn's patient no. ° 49 AIEC 4 8 nd nd Crohn's patient no. ° 51 AIEC 8 >8 nd nd Example 4 Post-treatment measurement of microbiome health for 5-temocilini and ciprofloxacin in mice Mice were treated with 30 mg / kg (by oral gavage) of either temocillin or ciprofloxacin. Mice were dosed once daily for 3 consecutive days after stable GI AIEC colonization. Fecal sediments from four mice (1–2 per mouse) were pooled, subjected to bacterial DNA enrichment, and analyzed using Illumina MiSeq 2. Bacterial abundance and diversity were quantified using 16S rRNA gene sequencing data, analyzed, and compared with untreated mice. Richness (Figure 2A, OTU count, p<0.05) and diversity (Figure 2B, Shannon index, p<0.05) in temocillin-treated mice were comparable to those in untreated (naïve) mice and superior to those treated with ciprofloxacin. The content of all references (including bibliographic references, the 15 issued patents, published patent applications, and jointly pending patent applications) cited in this application is expressly incorporated herein in its entirety by reference. Unless otherwise defined, all technical and scientific terms used herein have the meanings normally understood by a person skilled in the art.

Claims

1. A method for treating an intestinal disorder in a subject experiencing invasive adherent E. coli (IAEC) overgrowth of the intestine, comprising administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof.

2. The method of claim 1, wherein the bowel disorder is selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome associated with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, pouchitis, and diverticulitis.

3. The method of claim 1 or 2, wherein the intestinal disorder is Crohn's disease.

4. The method of any of claims 1 to 3, wherein the subject is administered a pharmaceutically acceptable salt of temocillin.

5. The method of any of claims 1 to 4, wherein temocillin or a pharmaceutically acceptable salt thereof is administered orally.

6. A method for modulating the overgrowth of invasive adherent E. coli (AIEC) in a subject in need, comprising the step of administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof.

7. The method of claim 6, wherein the excessive growth of AIEC is in the subject's intestine.

8. The method of claim 6 or 7, wherein the subject has an intestinal disorder.

9. The method of any one of claims 6 to 8, wherein the subject has an intestinal disorder selected from irritable bowel syndrome (IBS) (e.g., irritable bowel syndrome associated with diarrhea (d-IBS), irritable bowel syndrome with constipation (c-IBS), irritable bowel syndrome alternating with constipation and diarrhea), Crohn's disease, traveler's diarrhea, colitis, ulcerative colitis, and diverticulitis.

10. The method of any of claims 6 to 9, wherein the subject has Crohn's disease.

11. The method of any of claims 6 to 10, wherein the modulation comprises a reduction of the excessive growth of AIEC.

12. The method of any of claims 6 to 11, wherein temocillin or a pharmaceutically acceptable salt thereof is administered orally.

13. A method for reducing intestinal inflammation caused by invasive adherent E. coH overgrowth (IAEC) in a subject in need, comprising the step of administering to the subject an effective amount of temocillin, or a pharmaceutically acceptable salt thereof.

14. The method of claim 13, wherein the subject has Crohn's disease.

15. A method for reducing one or more of diarrhea, fever, fatigue, abdominal pain and cramps, weight loss, and anemia in a subject having Crohn's disease, comprising administering to the subject orally an effective amount of temocillin or a pharmaceutically acceptable salt thereof.

16. The method of claim 15, wherein the subject is experiencing an overgrowth of invasive adherent E. coli (AIEC) of the intestine.

17. The method of any of claims 13 to 16, wherein temocillin 15 or a pharmaceutically acceptable salt thereof is administered orally.