WO2022010352A1 - 5-hydroxyindole et ses analogues utilisés comme stimulants de la motilité intestinale - Google Patents

5-hydroxyindole et ses analogues utilisés comme stimulants de la motilité intestinale Download PDF

Info

Publication number
WO2022010352A1
WO2022010352A1 PCT/NL2021/050434 NL2021050434W WO2022010352A1 WO 2022010352 A1 WO2022010352 A1 WO 2022010352A1 NL 2021050434 W NL2021050434 W NL 2021050434W WO 2022010352 A1 WO2022010352 A1 WO 2022010352A1
Authority
WO
WIPO (PCT)
Prior art keywords
analog
htp
hydroxyindole
gut
gut motility
Prior art date
Application number
PCT/NL2021/050434
Other languages
English (en)
Inventor
Sahar EL AIDY
Barbora WACLAWIKOVA
Original Assignee
Rijksuniversiteit Groningen
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rijksuniversiteit Groningen filed Critical Rijksuniversiteit Groningen
Publication of WO2022010352A1 publication Critical patent/WO2022010352A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/10—Laxatives

Definitions

  • the invention relates to the field of pharmacology, in particular gastrointestinal pharmacology.
  • the invention relates to compounds that affect gut motility.
  • the invention also relates to the treatment of conditions associated with reduced gut motility, such as constipation. More in particular, the invention relates to 5-hydroxyindole and analogs of 5-hydroxyindole as stimulants of gut motility.
  • the gastrointestinal (GI) tract is home to trillions of microbes.
  • the gut microbiota produces a wide range of small bioactive molecules derived from various substrates, including dietary precursors and medications (Van Kessel et al., Nat. Commun. 2019, 31, 1-31; Donia et al., Science 2015, 349, 80).
  • Such microbial conversion represents a significant regulatory mechanism by which gut microbes can alter intestinal host physiology, including gastrointestinal motihty (Yano et al., Cell 2015, 161, 264-276; Reigstad et al., FASEB J. 2015, 29, 1395-1403).
  • tryptamine produced by bacterial decarboxylation of dietary tryptophan accelerates gastrointestinal transit by activating epithelial G-protein coupled receptors, serotonin receptor 4, and increasing anion-dependent fluid secretion in the proximal colon of mice (Bhattarai et al., Cell Host Microbe 2018, 23, 775-785). Therefore, gut microbiota-derived molecules appear to functionally link the microbiota activity to the host gastrointestinal motility.
  • Gastrointestinal motility is a tightly controlled system involving two main neurotransmitters, acetylchohne (ACh) and serotonin (5-HT), and their receptors, which are respectively located on smooth muscle cells and neuronal structures of the enteric nervous system (ENS) (Hansen, Physiol. Res. 2003, 52, 1-30).
  • ENS enteric nervous system
  • 5-HT is released from a subset of enteroendocrine cells, enterochromaffin cells (EC cells), to act on intrinsic primary afferent neurons (IPANs), followed by the release of several molecules, including acetylcholine.
  • Constipation is a common, debihtating motihty disorder affecting up to 27 % of the population (Sanchez et al., Can. J. Gastroenterol. 2011, 25, 11-15). Constipation is also often associated with colorectal cancer, Parkinson's disease, childhood attention-dehcit/hyperactivity disorder and autism spectrum disorder, as well as, mood disorders. Recently, the administration of 5-hydroxytryptophan (5-HTP) to a mouse model of depression resulted in a normalized total gastrointestinal transit time and increased colonic motihty (Israelyan et al., Gastroenterology 2019, 157, 507-521).
  • 5-hydroxytryptophan 5-hydroxytryptophan
  • 5-HTP also known as oxitriptan
  • 5-HTP is a naturally occurring amino acid, as well as a chemical precursor and intermediate metabolite of the essential amino acid L-tryptophan in the biosynthesis of serotonin (Bertrand et al., Auton. Neurosci. 2010, 153, 47-57).
  • 5-HTP as a food supplement, or drug with or without other medications for the treatment of a wide variety of conditions, including depression, fibromyalgia, binge eating associated with obesity, chronic headaches, and insomnia (Birdsall, Altern. Med. Rev.
  • 5-HT receptors and 5-HT antagonists.
  • Gershon et al., Experientia 1985, 41, 863-868 describes multiple molecules, including 5-hydroxyindole, that can displace serotonin from its neuronal binding sites identified in intestinal membranes isolated from rabbits, guinea pigs, and mice.
  • 5-HTP is a structural homologue of L-tryptophan, which is metabolized by gastrointestinal bacterial tryptophanase (TnaA) enzyme to produce indole (Snell, Adv. Enzymol. Relat. Areas Mol. Biol. 1975, 42, 287-329).
  • TnaA gastrointestinal bacterial tryptophanase
  • the present inventors recognized the need in the art for a stimulant of gut motihty having low toxicity and high effectivity. In particular, they aimed at providing novel means and methods for increasing contractility of the gut. Another objective is to provide a stimulant that accelerates the total gut transit time.
  • 5-HI 5-hydroxyindole
  • the invention is based on the judicious insight that 5-hydroxyindole and analogs thereof are able to stimulate motihty of the mammalian gut.
  • indole derivatives which preferably have an oxygen-containing substituent at selected positions on benzene moiety of the indole core, reduced gut motility can be treated.
  • the invention is directed to a method for the treatment of a condition associated with reduced gut motility, comprising administering to a subject in need thereof a pharmaceutically effective amount of 5-hydroxyindole (5-HI) or an analog thereof having the capability to increase gut motility.
  • a pharmaceutically effective amount of 5-hydroxyindole (5-HI) or an analog thereof having the capability to increase gut motility comprising administering to a subject in need thereof a pharmaceutically effective amount of 5-hydroxyindole (5-HI) or an analog thereof having the capability to increase gut motility.
  • Gut motility or gastrointestinal (GI) motility refers to the movements of the digestive system, and the transit of the contents within it.
  • GI motility disorders There are a variety of motility disorders that can affect the GI tract from the very top (esophagus) to the very bottom (colon and rectum).
  • motility disorders can affect the GI tract from the very top (esophagus) to the very bottom (colon and rectum).
  • Each part of the GI tract - esophagus, stomach, small intestine, and large intestine - has a unique function to perform in digestion, and each has a distinct type of motility and sensation. When motility or sensations are not appropriate for performing this function, symptoms occur.
  • analog as used in this disclosure includes compounds that are similar to 5-HI.
  • the analog is a structural analog of 5-HI.
  • Useful structural analogs of 5-HI are described in this disclosure.
  • the similarity between the compounds that are similar to 5-HI and 5-HI itself can be a result of a chemical reaction, thereby yielding a compound having a chemical structure which is similar to that of 5-HI.
  • the analog of 5-HI can have one or more substituents, preferably one substituent.
  • the substituent can be any atom or molecule, other than hydrogen.
  • the presence of an oxygen atom in the substituent was found to be surprisingly advantageous to reach a potent stimulatory effect on gut motility.
  • the analog of 5-HI may comprise an oxygen-containing substituent that is directly attached at the benzene moiety of the indole ring system.
  • the analog has a hydroxyl group or a methoxy group.
  • the one or more substituent(s) can be at the benzene ring of the analog.
  • the one or more substituent(s) can be at the 4-, 5-, 6-, and/or 7-position of the phenyl. It may be that the presence of one or more substituents at the phenyl group of the 5-HI analog, especially those that have an oxygen atom, tend to increase the stimulating effect the compounds have on gut motihty when used (as a medicament) in the treatment of a condition associated with reduced gut motility.
  • the substituent is at the 4-, 5-, or 6-position.
  • the substituent is preferably at the 5- or 6-position, and more preferably at the 5-position.
  • alkoxy includes any hydrocarbon group that is bonded through an oxygen atom.
  • the hydrocarbon group may comprise one or more heteroatoms, i.e., other than hydrogen, carbon, and oxygen.
  • alkyls that are bonded through an oxygen atom are meant.
  • the alkoxy group may be connected with its oxygen atom directly to the compound.
  • the alkoxy group can be linear, such as ethoxy, or branched, such as isopropoxy. In particular, the alkoxy group is linear.
  • the alkoxy group can have 1-3 carbon atoms, such 1 or 2 carbon atoms.
  • the alkoxy group is selected from the group consisting of methoxy or ethoxy.
  • the alkoxy group is a methoxy.
  • the 5-HI analog can be a compound of formula (I) wherein R 1 is hydrogen, methyl or ethyl, preferably hydrogen or methyl.
  • the OR 1 group can be at the 4-, 5-, 6-, or 7-position.
  • the group can be at the 4-, 6-, or 7-position, and R 1 may be hydrogen.
  • the group is at the
  • R 1 is hydrogen.
  • the group can be at the 4-, 5-, 6- or 7-position, and R 1 is hydrogen.
  • the group can be at the 4-, 5-, 6- or 7-position, and R 1 is hydrogen.
  • the group can be at the 4-, 5-, 6- or 7-position, and R 1 is hydrogen.
  • the group can be at the 4-, 5-, 6- or 7-position, and R 1 is hydrogen.
  • the group can be at the 4-, 5-, 6- or 7-position, and R 1 is hydrogen.
  • R 1 may be methyl.
  • the group is at the 5-position, and R 1 is methyl.
  • the analog is selected from
  • the analog is 5-MI, 6-HI or 7-HI. More preferably, the analog is 5-MI or 6-HI. Even more preferably, the analog is 5-MI.
  • 5-HI or an analog thereof can be administered in any conventional manner by any route where it is active or becomes active.
  • Administration can be systemic or oral.
  • Administration is preferably oral.
  • the modus of administration can depend on the condition to be treated.
  • the selection of the specific route of administration can be selected or adjusted by a clinician according to methods known to the clinician to obtain the desired clinical response.
  • 5-HI or the analog thereof is preferably formulated in accordance with routine procedures as a pharmaceutical adapted for administration to humans.
  • 5-HI or the analog thereof can be formulated by combining 5-HI or the analog with a pharmaceutically acceptable carrier, diluent or vehicle.
  • the invention also provides a pharmaceutical composition comprising a 5-HI analog as herein disclosed, and with a pharmaceutically acceptable carrier, diluent or vehicle.
  • the composition is formulated for oral administration.
  • phrases "pharmaceutically acceptable” refers to, for example, compounds, materials, compositions, and dosage forms which are, within the scope of the practitioner's medical judgment, suitable for use in contact with tissues of animals, in particular humans.
  • the term may also include the approval by a (governmental) regulatory agency for use in animals, and more in particular for use in humans.
  • Such pharmaceutically acceptable carriers enable 5-HI or the analog thereof to be formulated as, for example, solutions; sustained-release formulation; suppository; tablets; pills; dragees; capsules; emulsions; liquids; gels; syrups; caches; pellets; powders; granules; slurries; lozenges; aqueous; or oily suspensions for oral ingestion by the subject or patient to be treated.
  • Pharmaceutical preparations for oral use can be obtained by, e.g., adding a solid excipient, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragees.
  • Suitable excipients include, but are not limited to, fillers, such as sugars, including, but not limited to, lactose, sucrose, mannitol, and sorbitol; cellulose preparations, such as, but not limited to, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and polyvinyl pyrrolidone.
  • disintegrating agents can be added, such as, but not limited to, the cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate.
  • a composition formulated for the oral administration of 5-HI or an analog thereof can contain one or more optional agents, e.g., sweetening agents, such as fructose, aspartame, or saccharin; flavoring agents, such as peppermint, oil of wintergreen, or cherry; coloring agents; and preserving agents to provide a pharmaceutically palatable preparation.
  • the compositions may be coated to delay disintegration and absorption in the gastrointestinal tract, particularly in the gut, thereby providing a sustained action over an extended period of time.
  • Selectively permeable membranes surrounding an osmotically active driving compound are also suitable for the oral administration.
  • Oral compositions can include standard vehicles, such as mannitol; lactose; starch; magnesium stearate; sodium saccharine; cellulose; magnesium carbonate; etc. Such vehicles are suitably of pharmaceutical grade.
  • Dragees can be provided with suitable coatings.
  • suitable coatings can be used, which can optionally contain Arabic gum; talc; polyvinyl pyrrolidone; carbopol gel; polyethylene glycol; and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures.
  • Dyestuffs or pigments can be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.
  • Pharmaceutical preparations which can be used orally include, but are not limited to, push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol.
  • the push-fit capsules can contain the active ingredients in admixture with filler, such as lactose or binders, such as starches, and/or lubricants, such as talc or magnesium stearate and, optionally, stabilizers.
  • filler such as lactose or binders, such as starches, and/or lubricants, such as talc or magnesium stearate and, optionally, stabilizers.
  • the active compounds can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • suitable liquids such as fatty oils, liquid paraffin, or liquid polyethylene glycols.
  • stabilizers can be added.
  • 5-HI or an analog thereof as herein disclosed may be administered either alone or in combination (concurrently or serially) with other compounds, such as pharmaceutically acceptable compounds and/or prodrugs.
  • 5-HI or the analog thereof can be administered in combination with an antibiotic and/or other known drug compound, for example, for the treatment of a condition associated with reduced gut motility, such as one or more further contractile agent(s).
  • 5-HI or an analog thereof is preferably used in an amount or dosage which is pharmaceutically effective.
  • pharmaceutically effective amount refers to the amount of active compound, or pharmaceutical, that elicits the biological or medicinal response that is being sought in a tissue, system or subject by a researcher, veterinarian, medical doctor or other clinician (practitioner).
  • the pharmaceutic effect is dependent on the condition being treated, the biological effect desired, and/or the clinical result desired.
  • the pharmaceutic effect can be a decrease in the severity of symptoms associated with the condition and/or inhibition (partial or complete; at least in part) of the progression of the condition, or improved treatment, healing, prevention or elimination of a condition, or side-effects.
  • the amount needed to elicit the therapeutic response can be determined, for example, based on the species, age, health, size, gender, and/or sex of the subject. Optimal amounts can also be determined based on monitoring of the subject's response to treatment.
  • the dosage to be administered may depend on the characteristics of the subject being treated, e.g., the particular animal or human treated, age, weight, health, types of concurrent treatment, if any, and frequency of treatments, and can be easily determined by, e.g., a chnician.
  • the dosing for 5-HI or analog thereof can be used and adjusted depending on, e.g., the factors above.
  • the selection of the specific dose regimen can be selected, adjusted, or titrated by a clinician according to methods known in the art to obtain the desired clinical response.
  • the dosage amount of 5-HI or analog thereof that is pharmaceutically effective in, e.g., the treatment of a particular condition will depend on the nature and extent of the condition, and can be determined by standard clinical techniques.
  • in vitro or in vivo assays may be employed to help identify optimal dosage ranges.
  • the precise dose to be employed also typically depends on the route of administration, and the seriousness of the condition, and should be decided according to the judgment of the practitioner and the patient's condition(s).
  • Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems (animal models).
  • 5-HI or analog thereof may be used for a time sufficient to treat the condition.
  • 5-HI or analog thereof is administered in a dosage range of 0.1 mg per kg of body weight or more.
  • the dosage range can be 1000 mg/kg body weight or less.
  • the dosage range can be 5-900 mg/kg body weight, such as 20-800 mg/kg body weight, 30-700 mg/kg body weight, or 40-600 mg/kg body weight.
  • the dosage range is 0.1-100 mg/kg body weight, such as 20-90 mg/kg body weight, 30-80 mg/kg body weight, or 40-70 mg/kg body weight.
  • the dosage range is 1-50 mg/kg body weight, such as 10-40 mg/kg body weight, 15-35 mg/kg body weight, or 20-30 mg/kg body weight.
  • 5-HI or analog thereof or (pharmaceutical) compositions comprising 5-HI or analog thereof can be prepared in unit dosage form.
  • the composition can be divided into unit doses containing appropriate quantities of 5-HI or analog thereof.
  • the unit dosage form can be a packaged preparation, where the package contains discrete quantities of the preparations, e.g., packeted tablets, capsules, and powders in vials or ampules.
  • the unit dosage form can be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.
  • 5-HI or an analog thereof can be administered orally to the subject in need thereof.
  • the phrase "in need thereof' includes that a subject has been identified as having a need for the particular treatment. The identification of such a need can be by any means of diagnostics. In particular, in any of the methods and treatments described herein, the subject is in need thereof.
  • treatment and “treating” are not meant to be limited to curing. Treating also includes alleviating at least one symptom of a condition, removing at least one symptom of a condition, lessen at least one symptom of a condition, and/or delaying the course of a condition.
  • treatment includes methods of therapy and optionally diagnosis. Also, the terms include therapeutic treatment and prophylactic or preventative measures wherein the object is to prevent or slow down (lessen) an undesired physiological condition, or obtain beneficial or desired clinical results.
  • Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of condition, stabilized (i.e., not worsening) state of condition, delay in onset or slowing of condition progression, amelioration of the condition state or remission (whether partial or total), whether detectable or undetectable, an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition.
  • Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment may also include prolonging survival as compared to expected survival if not receiving treatment.
  • treatment of a condition associated with reduced gut motility preferably means an activity that prevents, alleviates and/or ameliorates reduced gut motility caused by the condition.
  • the subject can be any animal, including vertebrates, e.g., wild, domestic, and farm animals.
  • the subject is a human subject.
  • the subject may suffer from abnormal motility patterns in the small intestine can lead to symptoms of intestinal obstruction. Symptoms of bloating, pain, nausea, and vomiting can result either from weak contractions or from disorganized (unsynchronized) contractions that result from intestinal muscle (visceral myopathy) or nerve (visceral neuropathy) problems.
  • the symptoms of constipation are infrequent bowel movements [usually less than 3 per week], passage of hard stools, and sometimes difficulty in passing stools.
  • One motility problem that can lead to constipation is a decrease in the number of high amplitude propagating contractions (slow transit) in the large intestine. The test used to detect this is a transit time (Sitzmark) study.
  • 5-HI or analog thereof is advantageously used for treating a condition associated with reduced gut motility.
  • the condition can be, for example, constipation.
  • the constipation can be associated with, for example, colorectal cancer or Parkinson's disease.
  • 5-HI or analog thereof can stimulate release of 5-HT from EC cells and/or L-VDCCs on colonic smooth muscle cells.
  • 5-HI and analogs thereof, particularly those described in this disclosure can be used in medical applications, such as in medicine.
  • 5-HI and analogs thereof, particularly those described in this disclosure, thereof can be used in methods for therapy and/or (in vivo ) diagnostics.
  • the method for therapy and/or (in vivo ) diagnostics can be a method for detecting, preventing, and/or treating, preferably treating, a condition associated with reduced gut motihty, particularly constipation.
  • 5-HI and analogs thereof, particularly those described in this disclosure can be used in the preparation or manufacture of a medicament for detecting, preventing, and/or treating, preferably treating, one or more conditions associated with reduced gut motihty, particularly constipation.
  • 5-HI and analogs thereof, particularly those described in this disclosure, can be used for the manufacture of a medicament for the prevention or treatment, preferably treatment, of a condition associated with reduced gut motility, such as constipation.
  • the constipation may be associated with, for example, colorectal cancer or Parkinson's disease.
  • 5-hydroxyindole or an analog thereof for use in the treatment of a condition associated with reduced gut motihty, wherein a pharmaceutically effective amount of 5-HI or analog thereof having the capability to increase gut motihty is administered to a subject in need thereof.
  • the analog of 5-HI is preferably as described in this disclosure, such as a compound of formula (I).
  • the analog can be selected from 4-HI, 5-MI, 6-HI, and 7-HI.
  • the analog is selected from 5-MI, 6-HI and 7-HI.
  • the analog is 5-MI or 6-HI.
  • 5-HI or the analog thereof is preferably administered as described in this disclosure, especially orally, to a subject in need thereof as described in this disclosure.
  • the condition associated with reduced gut motility is preferably as described in this disclosure, in particular constipation.
  • 5-HI or the analog thereof may be administered in a dosage range of 0.1 mg per kg of body weight or more.
  • the dosage range can be 1000 mg/kg body weight or less.
  • the dosage range can be 5-900 mg/kg body weight, such as 20-800 mg/kg body weight, 30-700 mg/kg body weight, or 40-600 mg/kg body weight.
  • the dosage range is 0.1-100 mg/kg body weight, such as 20-90 mg/kg body weight, 30-80 mg/kg body weight, or 40-70 mg/kg body weight.
  • the dosage range is 1-50 mg/kg body weight, such as 10-40 mg/kg body weight, 15-35 mg/kg body weight, or 20-30 mg/kg body weight.
  • the treatment of conditions associated with reduced gut motility can be targeted to stimulate the release of 5-HT from EC cells.
  • the treatment may instead, or in addition to, activate L-VDCCs.
  • 5-HI or the analog thereof can stimulate the release of 5-HT from EC cells and/or activate L-VDCCs on colonic smooth muscle cells.
  • Any other known medicament, compound or composition used for the detection, prevention, and/or treatment of a condition associated with reduced gut motihty can be used in co-therapy, co- administration, or co-formulation with the 5-HI or the analog thereof.
  • 5-HI or the analog thereof for use as an activator of gut motility or as a contractile agent.
  • the analog may be as described in this disclosure, such as a compound of formula (I) or (II).
  • the analog may be 4-hydroxyindole, 6-hydroxyindole, 7-hydroxyindole, or 5-methoxyindole.
  • the analog is 6-hydroxyindole, 5-methoxyindole or 7-hydroxyindole. More preferably, the analog is 5-methoxyindole or 6-hydroxyindole. Even more preferably, the analog is 5-methoxyindole.
  • the analog can be a compound of formula (II), wherein the formula is R 1 can be amino, hydroxyl, methoxy, or ethoxy.
  • R 1 can be at the 4-, 5-, 6-, or 7-position.
  • R 1 may be hydroxyl and can be at the 4-, 6-, or 7-position, preferably at the 4- or 6-position.
  • R 1 may be methoxy and can be at the 4-, 5-, 6-, or 7-position, preferably at the 5-position.
  • R 1 can be amino and can be at the 5-position.
  • a 5-HI analog for use as a medicament.
  • the analog is a compound of formula (I), wherein the formula is
  • R 1 can be hydrogen or methyl.
  • the OR 1 group can be at the 4-, 5-, 6- or 7-position.
  • the group can be at the 4-, 6-, or 7-position, and R 1 may be hydrogen.
  • the group can be at the 4-, 5-, or 6-position, and R 1 may be methyl.
  • the analog is selected from
  • the analog is 5-MI, 6-HI or 7-HI. More preferably, the analog is 5-MI or 6-HI. Even more preferably, the analog is 5-MI.
  • kits comprising one or more containers filled with 5-HI or an analog thereof as described in this disclosure.
  • container(s) can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration for treating a condition as described in this disclosure, preferably a condition associated with reduced gut motility, such as constipation.
  • the kit may contain one or more analogs of 5-HI, such as
  • compositions including pharmaceutical compositions, comprising one or more of these compounds, are also provided.
  • These compounds may be used as medicaments, for example, as pharmaceuticals, and may be used in medical applications, such as in medicine.
  • These compounds may be used for treating (medical) conditions, especially conditions associated with reduced gut motility, such as constipation.
  • These compounds may be used in a method of therapy, in particular to detect and/or treat, preferably to treat, a condition associated with reduced gut motility, especially in the treatment of constipation which can be associated with, e.g., colorectal cancer or Parkinson's disease.
  • a pharmaceutical composition comprising a 5-HI analog as defined in this disclosure.
  • the 5-HI analog is a compound of formula (I).
  • the 5-HI analog is a compound of formula (I).
  • 5-HI analog is selected from 4-hydroxyindole, 5-methoxyindole,
  • the pharmaceutical composition may be suitable for oral administration.
  • the pharmaceutical composition may be in a form for oral administration as described in this disclosure.
  • the pharmaceutical composition may comprise a pharmaceutically acceptable carrier and/or one or more optional agents, as described in this disclosure.
  • the invention is directed to a method for identifying an activator of gut motility.
  • the method comprises contacting in vitro a 5-HI analog to be tested for activating gut motility with colonic tissue.
  • the method further comprises determining whether the compound induces muscle cell contractility. Either or both steps may be performed as described in this disclosure.
  • the invention is directed to a method for providing a compound having the capabihty to increase gut motihty.
  • the method comprises providing a hbrary of 5-HI analogs.
  • the method further comprises testing in vitro one or more members of the library, i.e., one or more of the 5-HI analogs, on their capacity to activate L-VDCCs.
  • the testing step may be performed as described in this disclosure.
  • Figure 1 Bacteria in fecal samples from healthy volunteers convert 5-HTP to 5-HI ex vivo.
  • Figure 2 Bacteria in fecal samples from healthy volunteers convert 5-HTP to 5-HI.
  • C,D Michaelis-Menten kinetic curves for C) 5-HTP and D) tryptophan as substrates for 200 nM or 20 nM TnaABW25ii3, respectively. Reactions were performed in biological triphcate using 5-HTP concentrations ranging from 0 to 12 mM and tryptophan concentrations ranging from 0 to 14.58 mM. Enzyme kinetics were calculated using a nonlinear Michaelis-Menten regression model for 5-HTP and a non-linear substrate inhibition kinetic model for tryptophan. Error bars represent the SEM.
  • F,G Overnight cultures of F. nucleatum subsp. animalis and F. simiae incubated anaerobically at 37 °C with 50 mM 5-hydroxytryptophan (5-HTP). Curves are a representative example of three biological replicates.
  • FIG. 4 Gut bacteria harboring tryptophanase are responsible for conversion of 5-HTP to 5-HI.
  • Curves represent one example of three biological rephcates.
  • Trp tryptophan
  • FIG. 5 Bacterial production of 5-HI is dependent on the microbiota composition and pH levels.
  • A) Microbial profiling of human fecal samples from High, Intermediate and Non-Converters shows decreasing relative abundance of inoA- encoding gut bacterial genera and increasing relative abundance of Bifidobacteria genera, respectively. Chromatograms on the left show variation in conversion of 5-HTP to 5 -HI after 24 hours of anaerobic incubation with 5-HTP.
  • FIG. 6 Bacterial production of 5-HI is dependent on the microbiota composition.
  • Pie charts represent the difference in relative percentage of TnaA-proteins in the various bacterial phyla represented in the intestinal tract (exploded parts) among High, Intermediate and Non-Converters. Parts annotated in light blue (not exploded) show different relative percentage of Bifidobacterium genera among High, Intermediate and Non-Converters. Parts annotated in shades of grey represent the rest of bacterial genera detected in the samples.
  • Figure 7 5-HI stimulates GI motility in vivo.
  • Data were analyzed using the RM 2-way ANOVA followed by Fisher's LSD test (*p ⁇ 0.05) (before and after treatment) and two-tailed unpaired t-test (*p ⁇ 0.05) was used for analysis of data from 5-HI-treated group and vehicle group after treatment. Error bars represent SEM.
  • FIG. 8 5-HI is a potent stimulator of gut contractility.
  • Bar graph represent a significantly increased 5-HT release, which is Ca 2+ influx dependent, from RIN 14B cells after stimulation with 100 mM of 5-HI. Data were analyzed using the two-tailed unpaired t-test ⁇ 0.0001).
  • H-K Bar graphs and illustrative recordings in the rat colon representing enhanced response of the ACh-induced contractility by 100 mM 5-HI (H), which was not inhibited after the addition of 1 mM AQ-RA 741 (I), but was significantly increased after adding 100 nM DAU 5884 hydrochloride (J), and significantly inhibited with 1 mM nifedipine (K).
  • B-E Bar graphs and illustrative recordings representing the inhibitory effect of either 1 mM ondansetron (B), 1 mM SB-207266 (C), 1 mM tetrodotoxin (TTX) (D) or 1 mM hexamethonium (E) on the 5-HT -induced response.
  • Bar graph represents no inhibitory effect of 1 mM hexamethonium on 5-HI -induced response.
  • FIG. 10 Proposed model for the mechanisms by which 5-HI or analog thereof accelerates intestinal motility.
  • 5-HT is released from EC cells to IPANs. Stimulation of IPANs initiate a signaling pathway ending up with release of neurotransmitters to excitatory cholinergic motor neurons in the ENS releasing ACh to the smooth muscle cells causing stimulation of muscarinic receptors 2 and 3 (M2/3R), which induce contractions that depend on voltage-dependent and -independent Ca 2+ entry followed by muscle contraction.
  • M2/3R muscarinic receptors 2 and 3
  • 5-HI translocates through the gut tissue, where it reaches the smooth muscle cells and accelerates gastrointestinal motihty directly via activation of L-VDCCs.
  • NSSCs non-selective Ca 2+ channels.
  • Figure 11 Dose response curves and ECso values of 5-hydroxyindole and analogs. Compounds were evaluated in an ex vivo rat colonic tissue model. 5-HI, 5-hydroxyindole; 6-HI, 6-hydroxyindole; 4-HI, 4-hydroxyindole; 5-MI, 5-methoxyindole; 5-AI, 5-aminoindole; 7-HI, 7-hydroxyindole.
  • FIG. 12 Overlay of dose response curves and EC50 values of 5-HI and analogs thereof with hydroxy group at different position.
  • Panel A 5-HI vs. 6-HI
  • Panel B 5-HI vs. 4-HI
  • Panel C 5-HI vs. 7-HI
  • Pane Pane
  • D 6- HI vs. 4-HI
  • Panel E 7-HI vs. 4-HI
  • Panel F 6-HI vs. 7-HI.
  • FIG. 13 Overlay of dose response curves and comparison of ECso values of 5-HI and analogs thereof having a different substituent at position 5.
  • Panel A 5-HI vs. 5-AI (5-aminoindole);
  • Panel B 5-HI vs. 5-MI (5-methoxyindole);
  • Panel C 5-AI vs. 5 -MI.
  • FIG. 14 5-HI and analogs and their effect on rat colonic contractility.
  • Panel A 5-HI.
  • Panel B 6-HI (6-hydroxyindole);
  • Panel C 4- MI (4-methoxyindole);
  • Panel D 5-AI (5-aminoindole);
  • Panel E 4-HI (4-hydroxyindole);
  • Panel F 7-HI (7-hydroxyindole);
  • Panel G 5-EI (5- ethoxyindole). All compounds were tested at 100 mM. Data were analyzed using the Wilcoxon matched-pairs (before/after) signed rank test (*p ⁇ 0.05; **p ⁇ 0.01; ****p ⁇ 0.0001). Error bars represent SEM.
  • Example 1 Gut bacteria convert 5-HTP to 5-HI
  • 5-HTP is absorbed throughout the entire GI tract (Jacobsen, et al., Neuropsychopharmacology 2016, 41, 2324-2334; Gijsman, et al., J. Clin. Psychopharmacol. 2002, 22, 183-189) when taken as a food supplement or as an antidepressant, where it is partially converted in the gut into serotonin before it reaches the brain (Bertrand (2010)).
  • 5-HTP is metabolized in the gut lumen before it is absorbed in the gut tissue and is converted into serotonin
  • five healthy volunteers were randomly selected out of the 18 subjects, who participated in the fecal samples' donation.
  • Subjects were orally administered 5-HTP tablets (Swanson Health Products, F argo, ND, USA) (50 mg 5-HTP per tablet) daily for 1 week, as recommended by the manufacturer.
  • Fecal samples were collected for targeted metabolomic analysis on Day 0 (control, no 5-HTP ingested), Day 1 (first ingestion of 5-HTP tablet), Day 3, and Day 8 (one day after last 5-HTP tablet was ingested), respectively.
  • Example 2 Bacterial tryptophanase responsible for converting 5-HTP
  • 5-HI is a structural analog of indole, which is produced by bacterial degradation of L-tryptophan via TnaA enzyme, it was hypothesized that 5-HTP is also a substrate for the TnaA enzyme (Snell (1975))
  • TnaA gene from E. coli BW25113 was expressed in E. coli BL21 (DE3) and then purified.
  • Table 1 Enzyme kinetics determined by non-linear Michaelis-Menten regression model for 5-HTP and non-linear substrate inhibition kinetic model for tryptophan.
  • E. coli BW25113 was used as a query to search the U.S. National Institutes of Health Human Microbiome Project (HMP) protein database.
  • HMP Health Human Microbiome Project
  • the proteins were considered homologous when the minimal identity percentage was above 30 % and query cover was above 90 %.
  • This analysis identified several genera containing TnaA; Escherichia, Fusobacterium, Bacteroides, Aeromonas, Clostridium, Klebsiella, Providencia and Propionibacterium ( Figure 3E).
  • TnaA is the enzyme involved in 5-HTP conversion to 5-HI, and this function is encoded by genomes of a variety of gut bacterial species that are abundantly present in both the small and large intestine (Jaglin, et al., Front. Neurosci. 2018, 12, 216).
  • Example 3 Bacterial production of 5-HI dependent on microbiota composition and pH
  • 5-HI Being the bacterial metabolite of 5-HTP, that has been recently shown to accelerate the gut motility (Israelyan (2019)), 5-HI was tested for its ability to also affect gut motility in vivo.
  • WTG Wild-type Groningen
  • TGTT total gut transit time
  • the 5-HI-treated group had significantly decreased TGTT compared to the baseline measurements before treatment (Figure 7 A). Moreover, the defecation frequency was significantly increased in the 5-HI-treated group compared to the vehicle group ( Figure 7B), without affecting food intake or changes in body weight (Figure 7C). Together, the in vivo data suggest that 5-HI stimulates the GI motihty in rats.
  • Example 5 - 5-HI is a potent stimulator of the colonic gut contractility
  • 5-HI can exert its stimulatory effect on gut motility observed in vivo through its stimulation of serotonin (5-HT) release from the EC cells or its action on enteric neurons, or smooth muscles underlying the wall of the gut.
  • 5-HT serotonin
  • TTX tetrodotoxin
  • the neurotransmitter ACh (50 mM) was applied to the tissue to induce a maximum increase in the intestinal smooth muscle tone (Tezuka et al., Exp. Gerontol. 2004, 39, 91-100), which has been long recognized as a key component in gut motility (Lentle et al., Neurogastroenterol. Motil. 2013, 25, 931-942).
  • addition of 100 mM 5-HI significantly intensified the amphtude of the ACh-induced contractility in the colonic tissue (Figure 8H).
  • the ACh response is mainly exerted through muscarinic acetylcholine receptor subtypes 2 (M2R) and 3 (M3R) located on the smooth muscle cells in the GI tract (Hansen (2003)).
  • M2R muscarinic acetylcholine receptor subtypes 2
  • M3R muscarinic acetylcholine receptor subtypes 2
  • AQ-RA 741 (1 mM) or DAU 5884 hydrochloride (100 nM)
  • selective antagonists of the M2R or M3R respectively (Doods et al., Eur. J. Pharmacol. 1991, 192, 147-152; Di Patrizio et al., Am.
  • the stimulation of muscarinic receptors by ACh causes the opening of a variety of cationic channels (e.g ., non-selective Ca 2+ channels (NSCCs), specifically the interconnected transient receptor potential channel 4 (TRPC4) and 6 (TRPC6) (Tsvilovskyy et al., Gastroenterology 2009, 137, 1415-1424) and L-type voltage-dependent Ca 2+ channels (L-VDCCs) (Wegener (2006))) in the smooth muscle cells of the GI tract, thereby producing Ca 2+ influx followed by muscle contraction.
  • NSCs non-selective Ca 2+ channels
  • TRPC4 and 6 TRPC6
  • L-VDCCs L-type voltage-dependent Ca 2+ channels
  • ACh was applied sequentially with 5-HI followed by the addition of selective antagonists of TRPC4 and TRPC6, ML 204 (1 mM) and SAR 7334 (1 mM), respectively (Tsvilovskyy (2009)). Only the ACh-induced response, but not the 5-HI -induced response, was inhibited ( Figures 91 and 9J), respectively.
  • 5-HI was applied sequentially with 1 mM nifedipine (Tsai et al., J. Ethnopharmacol. 2012, 142, 694-699), a non-selective antagonist of L-VDCCs.
  • the potency of 5-HI was identified, a product of gut microbial conversion of the dietary supplement and antidepressant 5-HTP, to accelerate GI motility indirectly via stimulation of 5-HT release from EC cells, and directly via activation of L-type calcium channels located on the colonic smooth muscle cells.
  • 5-HI is often wrongly associated with 5-hvdroxvoxindole (Lee (2010); Lee et al., Trends Microbiol. 2015, 23, 707-718), which is an oxindole with a hydroxyl group at position 5 of the indole ring, and is detected in blood, plasma, and the brain (Crumeyrolle-Arias et al., J. Neurosci. Res. 2008, 86, 202-207).
  • the primary outcome is that a wide variety of small and large intestinal bacteria possessing tryptophanase (TnaA) activity, typically converting tryptophan to indole, can efficiently convert 5-HTP into 5-HI in the intestinal lumen, with variability in the conversion levels ( Figures 1A, 3F, and 3G).
  • the variation in the production of 5-HI in the human fecal samples was associated with high relative abundance of Bifidobacteria in the human fecal samples ( Figure 5A), which is consistent with previous data showing that Bifidobacterium spp. isolated from healthy individuals inhibited the TnaA activity of E. coli or total human intestinal microbiota (Park et al., Arch. Pharm. Res. 1998, 21, 149-159).
  • TnaA activity and indole levels are reported to be decreased in the infant fecal samples, which are dominated by Bifidobacteria compared to fecal samples from adults (Ishibashi et al., Malays. J. Nutr. 1997, 3, 149-159).
  • the inhibition effect of Bifidobacteria on TnaA is due to the ability of Bifidobacteria to decrease the pH of the bacterial or fecal suspensions ( Figures 5B-5D), which is consistent with previous studies where TnaA activity is shown to be dependent on pH (Kim (1995)).
  • TnaA expression is strictly controlled via various regulatory networks, including catabolite repression and transcription attenuation mechanisms (Gong et al., Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 8997-9001;
  • L-VDCCs L-type Ca v 1.2 Ca 2+ channel
  • GI motihty disorders including constipation (Zhang (2010); Beyder et al., Therap. Adv. Gastroenterol 2012, 5, 5-21; Liu et al., Gastroenterology 2001, 120, 480-489).
  • L-VDCCs agonists may serve to restore intracellular Ca 2+ dependent motility to the constipated colon by activating the Ca 2+ channel (Liu (2001)). This claim is further strengthened by the results where the oral administration of 5-HI reduces TGTT in WTG rats ( Figure 7A).
  • the inventors arrived at a model for the mechanisms by which 5-HI is expected to accelerate intestinal motihty depicted in Figure 10.
  • the study protocol was evaluated by the ethical committee of the University of Groningen Medical Center. Eighteen volunteers (8 male and 10 female) between the ages 20 and 40 were recruited. Participants were excluded if they had used antibiotics, diarrhea inhibitors, laxatives, proton pump inhibitors, or had any gastrointestinal discomfort within the last three weeks. The study coordinator met with each eligible participant to review the consent and study details. All subjects signed the confidentiality and consent form.
  • a plastic collection container, relief container, and gloves were provided to each subject for fecal collection. Subjects collected the feces and delivered it immediately to the study coordinator who delivered it to the laboratory for processing. One part of the fecal specimen was homogenized in Liquid Amies medium containing 20 % glycerol (Table 2), snap-frozen and stored in -80 °C. The second part was only snap-frozen without any additives in liquid nitrogen and stored in -80 °C. Table 2. Constituents of Liquid Amies with 20 % medium.
  • the study protocol was evaluated by the ethical committee of the University of Groningen Medical Center. Five out of the 18 tested subjects who participated in the fecal samples donation, were randomly selected and asked to orally administer 5-HTP tablets (Swanson Health Products, Fargo, ND, USA) (50 mg 5-HTP per tablet) daily for one week, as recommended by the manufacturer. The study coordinator met with each participant to review the consent and study details. All subjects signed the confidentiality and consent form. Volunteers collected fecal samples on Day 0 (control, no 5-HTP ingested), Day 1 (first ingestion of 5-HTP tablet), Day 3 and Day 8 (one day after last 5-HTP was ingested), respectively.
  • a plastic collection container, relief container and 5-HTP tablets were provided to each subject for fecal collection on specified days. Subjects collected the feces and delivered it immediately to the study coordinator who delivered it to the laboratory for fecal samples extraction of metabolites. The rest of the sample was stored in -80 °C. Fecal samples extraction of metabolites
  • DNA extraction was performed using the Quick-DNATM Fecal/Soil Microbe Miniprep Kit (Zymo Research) according to manufacturer's instructions. Illumina 16S rRNA gene amphcon libraries were generated and sequenced at BaseClear (Leiden, The Netherlands). In short, barcoded amplicons from the V3-V4 region of 16S rRNA genes were generated using a 2-step PCR. 10-25 ng genomic (g)DNA was used as template for the first, PCR with a total volume of 50 pL using the 34 IF (5'-CCTACGGGNGGCWGCAG-3') and the 785R
  • PCR products were purified and the size of the PCR products were checked on Fragment analyzer (Advanced Analytical) and quantified by fluorometric analysis. Purified PCR products were used for the 2nd PCR in combination with sample -specific barcoded primers (Nextera XT index kit, Illumina). Subsequently, PCR products were purified, checked on a Fragment analyzer (Advanced Analytical) and quantified, followed by multiplexing, clustering, and sequencing on an Illumina MiSeq with the paired-end (2x) 300 bp protocol and indexing.
  • the sequencing run was analyzed with the Illumina CASAVA pipeline (vl.8.3) with demultiplexing based on sample -specific barcodes.
  • the raw sequencing data produced was processed removing the sequence reads of too low quahty (only "passing filter” reads were selected) and discarding reads containing adaptor sequences or PhiX control with an in-house filtering protocol.
  • a quality assessment on the remaining reads was performed using the FASTQC quality control tool version 0.10.0.
  • the Illumina paired reads were merged into single reads (so-called pseudoreads) through sequence overlap, after removal of the forward and reverse primers. Chimeric pseudoreads were removed and the remaining reads were ahgned to the RDP 16S gene databases.
  • the taxonomic depth of the lineage is based on the identity threshold of the rank; Species 99 %, Genus 97 %, Family 95 %, Order 90 %, Class 85 %, and Phylum 80 %.
  • Bacteria Escherichia coli DH5a and BL21 were routinely grown aerobically in Luria-Broth (LB) at 37 °C with continuous agitation. Other strains listed in Table 4 were grown either aerobically at 37 °C with continuous agitation or anaerobically at 37 °C in an EBB unless otherwise noted. Bacteria were inoculated from -80 °C stocks and grown overnight. Before the experiment, cultures were diluted 1:100 in fresh medium from overnight cultures. 5-HTP was supplemented during the lag or stationary phase depending on the experiment. Growth was followed by measuring the optical density (OD) at 600 nm in a spectrophotometer (UV1600PC, VWR International, Leuven, Belgium).
  • OD optical density
  • Fusobacterium nucleatum subsp. animalis DSM 19679 and Bifidobacterium breve DSM 20213 was grown anaerobically at 37 °C in an EBB. Fusobacterium nucleatum subsp. animalis and£>. breve was first restreaked on F astidious Anaerobe Agar supplemented with 5 % sheep blood or on MRS agar, respectively, and grown overnight before inoculation to liquid EBB culture. Before the experiment, 10 mL of B. breve was centrifuged and cells were washed in fresh 10 mL EBB. From the 100 %
  • B. breve suspension 50 % and 10 % of B. breve suspension was prepared.
  • 1% of the F. nucleatum subsp. animalis culture was inoculated.
  • F. nucleatum subsp. animalis was inoculated 1:100 to fresh medium, which served as a control.
  • Tryptophanase from E. coli BW25113 (TnaAE. Co/ ;BW25ii3, accession: CP009273) was amphfied using Phusion High-Fidelity DNA polymerase and primers listed in Table 5.
  • Amplified tnaA gene was cloned in pET15b, resulting in pBW002 (Table 6).
  • lysozyme 105281, Merck
  • DNase 11284932001, Roche Diagnostics
  • Ni-NTA nickel-nitrilotriacetic acid
  • Cell free extracts were loaded on 0.5 mL Ni-NTA matrixes and incubated on a roller shaker for 2 h at 4 °C.
  • reaction mixture was prewarmed for 5 min at 37 °C and was initiated by adding an enzyme solution and was terminated afterwards by the addition of 100 pL of 1 M HC1, following the addition of 100 pL of Kovac's reagent for indoles (60983, Sigma, The Netherlands). The supernatant was examined spectrophotometrically at 540 nm. The amounts of indole and 5-HI formed in each reaction were calculated from a standard curve. Michaelis-Menten kinetic curves were fitted using GraphPad Prism 7.
  • Tryptophanase amino acid sequence from E. coli BW25113 (NCBI accession: WP_001295247.1) was locally BLASTed against the protein sequences from the NIH HMP project (Bioproject ID 43021). All BLASTp hits were validated and only gut bacterial strains protein sequences were selected. The protein sequences were aligned in MEGA-X (Kumar et al.,
  • WTG rats Sixteen male adult wild-type Groningen (WTG) rats (Groningen breed, male, age: 19-23 weeks) housed 3-5 animals/cage had ad libitum access to water and food (Altromin 1414 mod. - NL_141005) in a temperature (21 ⁇ 1 °C) and humidity-controlled room (45-60 % relative humidity), with a 12 h light/dark cycle (lights off at 1:00 p.m.). These outbred rats are very frequently used in behavioral studies due to the high inter-individual variation (Koolhaas et al., J. Vis. Exp. 2013, 1-7), thus resembling, to some extent, the human inter-individual variation.
  • the 10 % sucrose solution was used as a vehicle for the 1.2 % carmine red (C1022, Sigma) and 0.5 % methylcellulose (M0512, Sigma) mixture. Rats were given 1.2 mL of the carmine red solution to measure total gut transit time before and after the treatment of 5-HI, for both groups, 5-HI-treated group as well as vehicle group. 5-HI treated group were given daily 30 mg/kg of 5-HI (H31859, Sigma) in drinking pipette with a 2.5 mL sucrose solution (10 % w/v) for a period of 11 days. 10 % Sucrose solvent was used only for in vivo experiment as described before (Van Kessel (2019)).
  • RIN14B cells (ATCC; (cat number: CRL-2059 LOT: 64037693) were seeded in 12-well plates at the rate of 2 x 10 5 cells/1 mL in RPMI1640 containing 10 % FBS/well and cultured for 72 h. The medium was removed before washing the cells with HBSS (+Ca 2+ , +Mg 2+ ) containing 0.1 % BSA and 2 mM fluoxetine (F132, Sigma) (HBSS-S). The HBSS-S was again removed and replaced with 0.25 mL HBSS-S containing 100 mM 5-HI or only 0.25 mL HBSS-S, after which the solution was incubated further for 30 min at 37 °C. The supernatants were collected and stored at -20 °C until 5-HT measurement using an enzyme immunoassay (EIA) kit (Abnova).
  • EIA enzyme immunoassay
  • Wild-type Groningen (WTG) male rats were sacrificed and a proximal colon was immediately removed and washed in IX PBS and placed in 0.7 % NaCl solution. Approximately 3 mm rings were cut and were placed in an organ bath (Tissue Bath Station with SSL63L force transducer, Biopac Systems Inc.
  • the (n) refers to the number of individuals, rats or rats' tissues used for each experiment.
  • the number of rats for in vivo experiment was based on previous study (Van der Wulp et al., J. Pediatr. Gastroenterol. Nutr. 2012, 55, 457-462) to achieve power of 80 % with a of 0.05. Specific test, significance and (n) number are indicated in the figure legends.
  • 5-HI Sigma Aldrich, H31859
  • 6-hydroxyindole (6-HI; Fluorochem, 023442)
  • 4-hydroxyindole (4-HI; Sigma Aldrich, 219878)
  • 5-MI 5-methoxyindole
  • 5-MI 5-methoxyindole
  • 7-hydroxyindole 7-HI; Sigma Aldrich, CDS005198
  • 5-aminoindole 5-AI; Sigma Aldrich, 307203
  • Indole Sigma Aldrich, W259306
  • indole-3-carboxaldehyde Sigma Aldrich, 129445
  • (lH-indol-3-yl)methanamine Fluorochem, 078831
  • 5-hydroxyindole-3-acetic acid 5-HIAA; Sigma Aldrich, H8876
  • Indole-3-acetic acid (Sigma Aldrich, 13750), 5-hydroxyoxindole (Sigma Aldrich, CDS004194), 5-hydroxy-2-carboxylic acid (Sigma Aldrich, 418608), indole-2 -carboxylic acid (TCI, 10332), and 1-methylindole (TCI, M0561) did not show any effect.
  • the six compounds were tested ex vivo on rat intestinal tissue (Organ-bath experiments; see above ) and the amplitude, and frequency of contractions were measured and compared to control (no treatment).
  • the half maximal effective concentration (EC50 values) were obtained and ranged from 33.5 mM for 5-MI, 41.4 mM for 5-HI, 57.5 mM for 4-HI, 116.4 mM for 6-HI, 158 mM for 7-HI, to 382 mM for 5-AI ( Figure 11).
  • the dose response curves provided an opportunity for the comparison of individual EC50 values and thus compare each molecule's effect on colonic contractility.
  • Potassium chloride is the natural contracting agent within the gut. Thus, it was considered that compounds that cause a stimulatory effect within the range of the maximal KC1 effect are safe to be applied in vivo. 100 mM of 5-HI, 6-HI, 4-HI, 5-MI, or 5-AI was applied subsequently with KC1 (30 mM).

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention porte sur le 5-hydroxyindole (5-HI) et ses analogues, et sur leur utilisation médicale, telle que le traitement d'affections associées à une diminution de la motilité intestinale. L'invention concerne une méthode pour le traitement d'une affection associée à une diminution de la motilité intestinale, consistant à administrer à un patient qui en a besoin une quantité pharmaceutiquement efficace de 5-HI ou d'un de ses analogues présentant la capacité d'augmenter la motilité intestinale.
PCT/NL2021/050434 2020-07-10 2021-07-09 5-hydroxyindole et ses analogues utilisés comme stimulants de la motilité intestinale WO2022010352A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202063050176P 2020-07-10 2020-07-10
US63/050,176 2020-07-10

Publications (1)

Publication Number Publication Date
WO2022010352A1 true WO2022010352A1 (fr) 2022-01-13

Family

ID=77300959

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL2021/050434 WO2022010352A1 (fr) 2020-07-10 2021-07-09 5-hydroxyindole et ses analogues utilisés comme stimulants de la motilité intestinale

Country Status (1)

Country Link
WO (1) WO2022010352A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7732479B2 (en) * 2004-08-19 2010-06-08 Tel Aviv University Future Technology Development L.P. Compositions for treating amyloid associated diseases
WO2014125084A1 (fr) * 2013-02-14 2014-08-21 Technische Universität München Dérivés d'hydroxyindalpine et leur utilisation médicale
WO2017161268A1 (fr) * 2016-03-18 2017-09-21 The Texas A&M University System Utilisation de métabolites du microbiote pour différencier des lymphocytes t naïfs et procédés associés d'induction ou de prévention des affections inflammatoires

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7732479B2 (en) * 2004-08-19 2010-06-08 Tel Aviv University Future Technology Development L.P. Compositions for treating amyloid associated diseases
WO2014125084A1 (fr) * 2013-02-14 2014-08-21 Technische Universität München Dérivés d'hydroxyindalpine et leur utilisation médicale
WO2017161268A1 (fr) * 2016-03-18 2017-09-21 The Texas A&M University System Utilisation de métabolites du microbiote pour différencier des lymphocytes t naïfs et procédés associés d'induction ou de prévention des affections inflammatoires

Non-Patent Citations (67)

* Cited by examiner, † Cited by third party
Title
BAE ET AL., BIOL. PHARM. BULL., vol. 33, 2010, pages 550 - 555
BANSAL ET AL., PROC. NATRL. ACAD. SCI., vol. 107, 2010, pages 228 - 233
BEYDER ET AL., THERAP. ADV. GASTROENTEROL, vol. 5, 2012, pages 5 - 21
BHATTARAI ET AL., CELL HOST MICROBE, vol. 23, 2018, pages 775 - 785
BIERSACK ET AL., CURR. DRUG TARGETS, vol. 13, 2012, pages 1705 - 1719
BOTSFORD ET AL., J. BACTERIOL., vol. 105, 1971, pages 303 - 312
CADENAS ET AL., FREE RADIC. RES. COMMUN., vol. 6, 1989, pages 11 - 17
CARUSO ET AL., J. INT. MED. RES., vol. 18, 1990, pages 201 - 209
CHEN ET AL., PHYSIOL. GENOMICS, vol. 31, 2007, pages 492 - 509
CHIMEREL ET AL., CELL REP., vol. 9, 2014, pages 1202 - 1208
CRUMEYROLLE-ARIAS ET AL., J. NEUROSCI. RES., vol. 86, 2008, pages 202 - 207
DI PATRIZIO ET AL., AM. J. PHYSIOL. -REGUL. INTEGR. COMP. PHYSIOL., no. 309, 2015, pages R805 - R813
DONIA ET AL., SCIENCE, vol. 349, no. 80, 2015
DOODS ET AL., EUR. J. PHARMACOL., vol. 192, 1991, pages 147 - 152
GERSHON ET AL., , BR. J. PHARMACOL. CHEMOTHER., vol. 29, 1967, pages 259 - 279
GERSHON ET AL., EXPERIENTIA, vol. 41, 1985, pages 863 - 868
GIJSMAN ET AL., J. CLIN. PSYCHOPHARMACOL., vol. 22, 2002, pages 183 - 189
GONG ET AL., PROC. NATL. ACAD. SCI. U.S.A., vol. 98, 2001, pages 8997 - 9001
HANSEN, PHYSIOL. RES., vol. 52, 2003, pages 1 - 30
HANSENSANDERS ET AL., NAT. REV. GASTROENTEROL. HEPATOL., vol. 9, 2012, pages 633 - 645
ISHIBASHI ET AL., MALAYS. J. NUTR., vol. 3, 1997, pages 149 - 159
ISRAELYAN ET AL., GASTROENTEROLOGY, vol. 157, 2019
JACOBSEN ET AL., NEUROPSYCHOPHARMACOLOGY, vol. 41, 2016, pages 2324 - 2334
JAGLIN ET AL., FRONT. NEUROSCI., vol. 12, 2018, pages 216
JEPERSEN ET AL., J. VIS. EXP., no. 1-9, 2015
KIM ET AL., ARCH. PHARM. RES., vol. 18, 1995, pages 351 - 355
KIM ET AL., J. CLIN. INVEST., vol. 108, 2001, pages 1051 - 1059
KOOLHAAS ET AL., J. VIS. EXP., 2013, pages 1 - 7
KOOYMAN ET AL., , BR. J. PHARMACOL., vol. 112, 1994, pages 541 - 546
KUMAR ET AL., CELL HOST MICROBE, vol. 28, 2020, pages 1 - 13
KUMAR ET AL., MOL. BIOL. EVOL., vol. 35, 2018, pages 1547 - 1549
LEE ET AL., FEMS MICROBIOL. REV., vol. 34, 2010, pages 426 - 444
LEE ET AL., TRENDS MICROBIOL., vol. 23, 2015, pages 707 - 718
LENTLE ET AL., NEUROGASTROENTEROL. MOTIL., vol. 25, 2013, pages 931 - 942
LETUNIC ET AL., NUCLEIC ACIDS RES., vol. 47, 2019, pages W256 - W259
LIU ET AL., GASTROENTEROLOGY, vol. 120, 2001, pages 480 - 489
LUMMIS ET AL., J. BIOL. CHEM., vol. 287, 2012, pages 40239 - 40245
LUND ET AL., MOL. METAB., vol. 11, 2018, pages 70 - 83
MANNAIONI ET AL., BR. J. PHARMACOL., vol. 138, 2003, pages 245 - 253
MCWILLIAMS ET AL., DRUGS, vol. 72, 2012, pages 2167 - 2175
MEDVEDEV ET AL., BIOL. TARGETS THER., vol. 1, 2007, pages 151 - 162
MULLER ET AL., CELL, vol. 158, 2014, pages 300 - 313
NARDINI ET AL., INT. J. CLIN. PHARMACOL. RES., vol. 3, 1983, pages 239 - 250
NEUROPSYCHOPHARMACOLOGY, vol. 44, 2019, pages 2082 - 2090
NOZAWA ET AL., PROC. NATL. ACAD. SCI. U.S.A., vol. 106, 2009, pages 3408 - 3413
PARK ET AL., ARCH. PHARM. RES., vol. 21, 1998, pages 149 - 159
PEREZ ET AL., FRONT. PHARMACOL, vol. 10, 2019, pages 1 - 15
RACKE ET AL., BEHAV. BRAIN RES., vol. 73, 1995, pages 83 - 87
REIGSTAD ET AL., FASEB J., vol. 29, 2015, pages 1395 - 1403
SANCHEZ ET AL., CAN. J. GASTROENTEROL., vol. 25, 2011, pages 11 - 15
SASAKI-IMAMURA ET AL., APPL. ENVIRON. MICROBIOL., vol. 76, 2010, pages 4260 - 4268
SIKANDER ET AL., CLIN. CHIM. ACTA, vol. 403, 2009, pages 47 - 55
SNELL, ADV. ENZYMOL. RELAT. AREAS MOL. BIOL., vol. 42, 1975, pages 287 - 329
TEZUKA ET AL., EXP. GERONTOL., vol. 39, 2004, pages 91 - 100
TSAI ET AL., J. ETHNOPHARMACOL., vol. 142, 2012, pages 694 - 699
TSVILOVSKYY ET AL., GASTROENTEROLOGY, vol. 137, 2009, pages 1415 - 1424
TULADHAR ET AL., BR. J. PHARMACOL., vol. 131, 2000, pages 1716 - 1722
UCHIYAMA ET AL., J. SMOOTH MUSCLE RES., vol. 40, 2004, pages 237 - 247
UNNO ET AL., BR. J. PHARMACOL., vol. 146, 2005, pages 98 - 108
UNNO ET AL., J. PHYSIOL., vol. 484, 1995, pages 567 - 581
VAN DER WULP ET AL., J. PEDIATR. GASTROENTEROL. NUTR., vol. 55, 2012, pages 457 - 462
VAN KESSEL ET AL., NAT. COMMUN., vol. 31, 2019, pages 1 - 31
WACLAWIKOVÁ BARBORA ET AL: "Gut bacteria-derived 5-hydroxyindole is a potent stimulant of intestinal motility via its action on L-type calcium channels", PLOS BIOLOGY, 1 January 2021 (2021-01-01), United States, pages e3001070, XP055853046, Retrieved from the Internet <URL:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7857600/pdf/pbio.3001070.pdf> [retrieved on 20211020], DOI: 10.1371/journal.pbio.3001070 *
WARDLE ET AL., , BR. J. PHARMACOL., vol. 118, 1996, pages 665 - 670
WEGENER ET AL., FASEB J., vol. 20, 2006, pages 1260 - 1262
YANO ET AL., CELL, vol. 161, 2015, pages 264 - 276
ZHANG ET AL.: "Neurogastroenterol", MOTIL, 2010, pages 22

Similar Documents

Publication Publication Date Title
Tian et al. Bifidobacterium with the role of 5-hydroxytryptophan synthesis regulation alleviates the symptom of depression and related microbiota dysbiosis
Gershon et al. The serotonin signaling system: from basic understanding to drug development for functional GI disorders
KR102425303B1 (ko) 항-메탄생성 조성물 및 그것의 용도
Jacob et al. Endogenous psychoactive tryptamines reconsidered: an anxiolytic role for dimethyltryptamine
Wei et al. Gut microbiota dysbiosis in functional gastrointestinal disorders: Underpinning the symptoms and pathophysiology
Waclawiková et al. Gut bacteria-derived 5-hydroxyindole is a potent stimulant of intestinal motility via its action on L-type calcium channels
JP2020523313A (ja) Ｍｅｇａｓｐｈａｅｒａ属の細菌株を含む組成物及びその使用
CN113382740A (zh) 包含细菌菌株的组合物
TWI787272B (zh) 包含細菌菌株之組合物
Barbaro et al. Escherichia coli Nissle 1917 restores epithelial permeability alterations induced by irritable bowel syndrome mediators
JP6884889B2 (ja) 細菌株を含む組成物
US20230398093A1 (en) Anti-methanogenic lovastatin analogs or derivatives and uses thereof
CN111107859A (zh) 包含细菌菌株的组合物
TW201907927A (zh) 包含細菌菌株之組合物
WO2005115471A2 (fr) Procedes et compositions pour le traitement de la dependance a la nicotine et de la demence
CN114615971A (zh) 包含细菌菌株的组合物
Legan et al. Tryptophan‐synthesizing bacteria enhance colonic motility
Forcén et al. Toll‐like receptors 2 and 4 exert opposite effects on the contractile response induced by serotonin in mouse colon: role of serotonin receptors
WO2022010352A1 (fr) 5-hydroxyindole et ses analogues utilisés comme stimulants de la motilité intestinale
WO2008011016A2 (fr) Méthodes de traitement du reflux gastrooesophagien
Nagakura et al. The selective 5-hydroxytryptamine (5-HT) 4-receptor agonist RS67506 enhances lower intestinal propulsion in mice
US20060258732A1 (en) Treatment and prevention of gastrointestinal disease using antagonists or partial agonists of 5HT1a receptors
Gao et al. Mucosal serotonin reuptake transporter (SERT) expression in IBS is modulated by gut microbiota via mast cell–prostaglandin E2
van Kessel et al. Gut bacterial deamination of residual levodopa medication for Parkinson’s
WO2023102450A1 (fr) Procédés de surveillance de l&#39;impact d&#39;une thérapie de transfert de microbiote dans un trouble du spectre autistique (tsa)

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21752768

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21752768

Country of ref document: EP

Kind code of ref document: A1