WO2025208014A1 - Vaccin conjugué haptène contre la xylazine - Google Patents

Vaccin conjugué haptène contre la xylazine

Info

Publication number
WO2025208014A1
WO2025208014A1 PCT/US2025/021994 US2025021994W WO2025208014A1 WO 2025208014 A1 WO2025208014 A1 WO 2025208014A1 US 2025021994 W US2025021994 W US 2025021994W WO 2025208014 A1 WO2025208014 A1 WO 2025208014A1
Authority
WO
WIPO (PCT)
Prior art keywords
xylazine
hapten
composition
administering
vaccine
Prior art date
Legal status (The legal status 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 status listed.)
Pending
Application number
PCT/US2025/021994
Other languages
English (en)
Inventor
Kim D. Janda
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Scripps Research Institute
Original Assignee
Scripps Research Institute
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 Scripps Research Institute filed Critical Scripps Research Institute
Publication of WO2025208014A1 publication Critical patent/WO2025208014A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/385—Haptens or antigens, bound to carriers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55505—Inorganic adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55516—Proteins; Peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55544—Bacterial toxins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/44—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • the present invention relates to compositions and methods for vaccines. More specifically, the present invention relates to vaccines against xylazine.
  • Xylazine is FDA-approved for veterinary use in animals as a sedative and pain reliever. Despite not being intended for human use, there is a growing popularity of individuals exposed to fentanyl, heroin, and other illicit drugs adulterated with xylazine. This phenomenon has prompted the Drug Enforcement Administration (DEA) to issue a public safety alert on March 20, 2023, reporting the widespread threat of fentanyl mixed with xylazine. Disturbingly, the DEA's findings reveal that approximately 23% of seized fentanyl powder and 7% of fentanyl pills contained xylazine.
  • DEA Drug Enforcement Administration
  • xylazine has acquired the nickname “zombie drug.”
  • Xylazine is a weak base, that rapidly penetrates the blood-brain barrier and due to its lipophilic nature it also has a rapid onset of action undergoing distribution to different tissues.
  • the pharmacology behind xylazine is that it primarily functions as an a2-adrenergic agonist.
  • Opioid vaccines have been developed for many different compounds, such as fentanyl, that use a hapten of the opioid conjugated to a carrier.
  • fentanyl that use a hapten of the opioid conjugated to a carrier.
  • U.S. Patent Application Publication No. 2022/0081400 to Janda, et al. discloses vaccines of fentanyl haptens and carfentanil haptens for use in treating opioid use disorder.
  • the present invention provides for the present invention provides for a composition of a xylazine hapten.
  • the present invention also provides for a composition of a xylazine hapten conjugate including the xylazine hapten conjugated to a carrier.
  • the present invention provides for a vaccine including the xylazine hapten or the xylazine hapten conjugate and a pharmaceutically acceptable carrier.
  • the present invention provides for a method of treating a substance use disorder, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine to an individual suffering from substance use disorder, and treating the individual.
  • the present invention provides for a method of preventing an overdose of xylazine or related compounds, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine to an individual at risk of an overdose, and preventing an overdose in the individual.
  • the present invention also provides for a method of generating antibodies against xylazine in an individual, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine to an individual, and generating antibodies in the individual.
  • the present invention provides for a method of treating an individual who has consumed a substance laced with xylazine, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine to an individual, and treating the individual.
  • FIGURE 1A shows the chemical structure of xylazine and FIGURE IB shows a conjugate of the xylazine hapten of Formula (I) for a vaccine study.
  • FIGURE 2 shows the synthetic route of a xylazine hapten of Formula (I).
  • FIGURE 3A is an NMR spectrum of the xylazine hapten of Formula (I).
  • FIGURE 3B is an NMR spectrum of the xylazine hapten of Formula (I).
  • FIGURE 4 shows a chemical structure of a side product in the synthesis of the xylazine hapten of Figure (I).
  • FIGURE 5A is a graph of LC-MS of condensation with formic acid during activation
  • FIGURE 5B is a graph of LC-MS of Lossen-rearrangement polymerization.
  • FIGURE 6 is a reaction scheme of side reactions in the synthesis of the xylazine hapten of Figure (I).
  • FIGURE 7 is an LC-MS graph showing side product Hapten-NHS-sulfo.
  • FIGURE 8A is a MALDI-TOF graph of BSA conjugate
  • FIGURE 8B is a MALDI-TOF graph of CRM197 conjugate.
  • FIGURE 9 is a graph of hapten BSA-conjugate density measurements.
  • FIGURE 10 is an illustration of an immunization scheme using xylazine hapten vaccines.
  • FIGURE 12 is a graph of dose-response curves for xylazine-induced respiratory depression.
  • FIGURE 13 is a standard curve of blood-brain biodistribution.
  • FIGURE 14 is a graph showing titer response of the xylazine vaccine with three different carrier proteins.
  • FIGURE 15A is a graph of locomotion in vaccine groups
  • FIGURE 15B is a graph of percentage of minute volume (MV) change post-drug challenge at 2 mg/kg.
  • FIGURE 16 is a graph showing blood-brain biodistribution of xylazine at 1 mg/kg, 15 minutes post-drug challenge.
  • FIGURE 17 is a reaction scheme for the synthesis of a xylazine hapten.
  • the present invention generally provides for compositions and methods for treating substance use disorder and overdose of xylazine in an individual with vaccines.
  • a "hapten” refers to a small molecule compound that is capable of generating or inducing the production of antibodies.
  • a “hapten conjugate” refers to a hapten conjugated to a larger carrier such as a protein that is capable of generating or inducing the production of antibodies.
  • the term "vaccine” refers to a composition that can provide active acquired immunity to and/or therapeutic effect (e.g. treatment) of a particular drug, disease, or pathogen.
  • a vaccine typically contains one or more agents that can induce an immune response in an individual against a drug, pathogen, disease, i.e. a target drug, pathogen, or disease.
  • the immunogenic agent stimulates the body's immune system to recognize the agent as a threat or indication of the presence of the target pathogen or disease, thereby inducing immunological memory so that the immune system can more easily recognize and destroy and/or eliminate any of the drug or pathogen on subsequent exposure.
  • Vaccines can be prophylactic (e.g. preventing or ameliorating the effects of a future drug overdose) or therapeutic (e.g., treating substance use disorder in an individual).
  • the administration of vaccines is referred to vaccination.
  • a vaccine can include a hapten or a hapten-conjugate.
  • adjuvant or “vaccine adjuvant” or “pharmaceutically acceptable adjuvant” refer to compounds used in a vaccine to enhance (e.g., increase, accelerate, prolong, and/or target) the specific immune response to the vaccine antigen/conjugate/hapten in order to enhance the individual's immune response to the vaccine.
  • the present invention provides for a composition of a xylazine hapten, such as shown in FIGURE IB or in FIGURE 17.
  • the xylazine hapten can include a xylazine epitope linked with a linker at the para-position of the dimethyl phenol amino ring with a three carbon spacer. It can be preferred to add a linker at a methyl group on an aromatic ring of xylazine.
  • FIGURE 17 shows a method of synthesis of a xylazine hapten that produces a linker on one of the methyl groups of the benzene ring.
  • the hapten can also include an analog or derivative of xylazine as the epitope, or a structurally similar compounds such as from the general class of phenothiazines.
  • the xylazine hapten can also be deuterated at one or more hydrogen positions to improve an immune response.
  • the present invention also provides for a composition of a xylazine hapten conjugate including the xylazine hapten conjugated to a carrier.
  • the carrier can be, but is not limited to, OVA (ovalbumin), BSA (bovine serum albumin), KLH (keyhole limpet hemocyanin), DT (diphtheria toxoid), CTB (cholera toxin subunit B), TT (tetanus toxoid), or CRM197. Conjugation of the hapten to the carrier can be performed by one skilled in the art.
  • the present invention provides for a vaccine including the xylazine hapten or the xylazine hapten conjugate and a pharmaceutically acceptable carrier.
  • the vaccine can include at least one adjuvant, such as, but not limited to, aluminum salts (alum), MF59, AS01, AS03, AS04, dmLT, LTA1, squalene based oil-in-water, MPLA-SM, flagellin derivatives, calcium salts, iron salts, zinc salts, acylated tyrosine, acylated sugars, cationically or anionically derivatized saccharides, polyphosphazenes, biodegradable microspheres, monophosphoryl lipid A (MPL), lipid A derivatives, 3-0-deacylated MPL, quil A, saponin based adjuvants (TQL 1055, QS-21), tocol, Freund's Incomplete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant
  • cytokines such as interleukins
  • the adjuvant can preferably be alum and CpG.
  • the compound of the present invention is administered and dosed in accordance with good medical practice, taking into account the clinical condition of the individual patient, the site and method of administration, scheduling of administration, patient age, sex, body weight and other factors known to medical practitioners.
  • the pharmaceutically "effective amount" for purposes herein is thus determined by such considerations as are known in the art. The amount must be effective to achieve improvement including but not limited to improved survival rate or more rapid recovery, or improvement or elimination of symptoms and other indicators as are selected as appropriate measures by those skilled in the art.
  • Sterile injectable solutions can be prepared by incorporating the compounds utilized in practicing the present invention in the required amount of the appropriate solvent with various of the other ingredients, as desired.
  • a pharmacological formulation of the present invention can be administered to the patient in an injectable formulation containing any compatible carrier, such as various vehicle, adjuvants, additives, and diluents; or the compounds utilized in the present invention can be administered parenterally to the patient in the form of slow-release subcutaneous implants or targeted delivery systems such as monoclonal antibodies, vectored delivery, iontophoretic, polymer matrices, liposomes, and microspheres.
  • any compatible carrier such as various vehicle, adjuvants, additives, and diluents
  • the compounds utilized in the present invention can be administered parenterally to the patient in the form of slow-release subcutaneous implants or targeted delivery systems such as monoclonal antibodies, vectored delivery, iontophoretic, polymer matrices, liposomes, and microspheres.
  • Examples of delivery systems useful in the present invention include: 5,225,182; 5,169,383; 5,167,616; 4,959,217; 4,925,678; 4,487,603; 4,486,194; 4,447,233; 4,447,224; 4,439,196; and 4,475,196. Many other such implants, delivery systems, and modules are well known to those skilled in the art.
  • the present invention provides for a method of treating a substance use disorder, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine as described above to an individual suffering from substance use disorder, and treating the individual.
  • Treating the individual can include generating antibodies against xylazine, analogs thereof, derivatives thereof, or structurally similar compounds.
  • the method can include reducing any effect that the xylazine has on the individual’s body, reducing and/or preventing addiction of the xylazine, or reduce cravings for the xylazine.
  • the present invention provides for a method of preventing an overdose of xylazine or related compounds, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine as described above to an individual at risk of an overdose, and preventing an overdose in the individual.
  • Preventing an overdose can include generating antibodies against xylazine, analogs thereof, derivatives thereof, or structurally similar compounds.
  • the compositions of the present invention can provide an antibody effect strong enough such that the individual does not die after ingesting xylazine, and so that additional medical attention can be sought if needed.
  • the present invention also provides generally for a method of generating antibodies against xylazine in an individual, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine as described above to an individual, and generating antibodies in the individual.
  • Antibody generation can be confirmed by taking a sample (such as a blood sample) from the individual and performing an assay (such as an ELISA).
  • the hapten, hapten conjugate, or vaccine can be administered in combination with an opioid hapten, opioid hapten conjugate, or opioid vaccine, or a vaccine for any other abused substance.
  • This can be useful in treating an individual who has consumed an opioid such as fentanyl, carfentanyl, morphine, heroin, codeine, oxycodone, or hydrocodone laced with xylazine. Therefore, the present invention provides for a method of treating an individual who has consumed a substance laced with xylazine, by administering the xylazine hapten, xylazine hapten conjugate, or vaccine as described above to an individual, and treating the individual. Treating the individual can include generating antibodies against xylazine, analogs thereof, derivatives thereof, or structurally similar compounds as well as against the opioid or other abused substance.
  • Antibodies offer advantages over small molecule antagonist treatments as they directly target the drug itself, bypassing complexities at the receptor level. Moreover, antibodies demonstrate higher safety profiles and selectivity against the targeted drug, making them a viable emergency measure for drug overdose, even in cases where the presence of drug mixture is uncertain. The mechanism of action underlying antibody therapy allows for its potential application across different drugs, thereby streamlining the development of therapeutics against emerging substances and those with multifaceted receptors like xylazine.
  • Embodiment 1 A composition comprising a xylazine hapten and optionally comprising an adjuvant.
  • Embodiment 2. The composition comprising a xylazine hapten of Embodiment 1 having the Formula (I)
  • Embodiment 3 The composition of either Embodiment 1 or Embodiment 2, wherein the xylazine hapten is conjugated to a pharmaceutically acceptable protein or peptide carrier.
  • Embodiment 4 The composition of Embodiment 3, wherein the protein carrier is KLH.
  • Embodiment 5 The composition of Embodiment 3, wherein the protein carrier is TT.
  • Embodiment 6 The composition of Embodiment 3, wherein the protein carrier is CRM197.
  • Embodiment 7 A vaccine comprising the xylazine hapten composition of Embodiment 1.
  • Embodiment 8 A vaccine comprising the xylazine hapten composition of Embodiment 2.
  • Embodiment 9 A vaccine comprising the xylazine hapten composition of Embodiment 3.
  • Embodiment 10 The vaccine of Embodiment 9, wherein the carrier protein is KLH.
  • Embodiment 11 The vaccine of Embodiment 9, wherein the carrier protein is TT.
  • Embodiment 12 The vaccine of Embodiment 9, wherein the carrier protein is
  • Embodiment 13 A method of preventing an overdose of xylene comprising administering the xylazine hapten composition of Embodiment 1 to a subject in need thereof.
  • Embodiment 14 A method of preventing an overdose of xylene comprising administering the xylazine hapten composition of Embodiment 2 to a subject in need thereof.
  • Embodiment 15 A method of preventing an overdose of xylene comprising administering the xylazine hapten composition of any one of Embodiments 3-6 to a subject in need thereof.
  • Embodiment 16 A method of preventing an overdose of xylene comprising administering the xylazine hapten vaccine of any one of Embodiments 7-12 to a subject in need thereof.
  • Embodiment 17 A method of treating a substance use disorder comprising administering the xylazine hapten composition of Embodiment 1 to a subject in need thereof.
  • Embodiment 18 A method of treating a substance use disorder comprising administering the xylazine hapten composition of Embodiment 2 to a subject in need thereof.
  • Embodiment 19 A method of treating a substance use disorder comprising administering the xylazine hapten composition of any one of Embodiments 3-6 to a subject in need thereof.
  • Embodiment 20 A method of treating a substance use disorder comprising administering the xylazine hapten vaccine of any one of Embodiments 7-12 to a subject in need thereof.
  • Embodiment 21 A method of generating antibodies against xylazine comprising administering the xylazine hapten composition of Embodiment 1 to a subject in need thereof.
  • Embodiment 22 A method of generating antibodies against xylazine comprising administering the xylazine hapten composition of Embodiment 2 to a subject in need thereof.
  • Embodiment 23 A method of generating antibodies against xylazine comprising administering the xylazine hapten composition of any one of Embodiments 3-6 to a subject in need thereof.
  • Embodiment 24 A method of generating antibodies against xylazine comprising administering the xylazine hapten vaccine of any one of Embodiments 7-12 to a subject in need thereof.
  • Embodiment 25 A method of treating consumption of a substance laced with xylazine comprising administering the xylazine hapten composition of Embodiment 1 to a subject in need thereof.
  • Embodiment 27 A method of treating consumption of a substance laced with xylazine comprising administering the xylazine hapten composition of any one of Embodiments 3-6 to a subject in need thereof.
  • Embodiment 28 A method of treating consumption of a substance laced with xylazine comprising administering the xylazine hapten composition of any one of Embodiments 7-12 to a subject in need thereof.
  • the hapten could not be preactivated in DMF for extended periods. Instead, activation was achieved by mixing the hapten with EDC/NHS-sulfo in H2O for 15 minutes before adding the carrier protein solution. The reaction then proceeded at room temperature for 2 hours. Mass studies revealed the occurrence of a side reaction between the hapten and NHS in the presence of EDC as a result of an elongated activation, resulting in the formation of the Lessen rearrangement side product, shown in FIGURE 4. The resulting amine could further react with another NHS molecule, leading to the formation of a series of polymers at higher EDC and NHS concentrations and longer incubation times.
  • FIGURE 6 shows a proposed reaction scheme.
  • FIGURE 7 is a graph showing side product ⁇ 1: 10.
  • Haptens are covalently linked to protein or peptide carriers, enabling their recognition by T-cell receptors wherein they are processed by antigen-presenting cells, loaded on to MHCs ultimately eliciting a specific immune response against the hapten.
  • protein carriers including OVA, BSA, KLH, DT (diphtheria toxoid), CTB (cholera toxin subunit B), TT (tetanus toxoid), and CRM197, a nontoxic mutant of DT. These carriers vary in protein size, cost, and immunogenicity.
  • the two least effective carriers, OVA and BSA have modest immunogenicity and are typically used in ELISA coatings for determining hapten-specific titer responses.
  • KLH has been extensively studied in preclinical practice, along with other evaluated options such as recombinant CTB. However, KLH has yet to be approved due to the complexity in its multi-subunit structure.
  • DT, TT, and CRM197 on the other hand have already gained approval for clinical use in polysaccharide conjugate vaccines. Previous comparison between these three carriers uncovered a trend in immunogenicity in the following orders: KLH ⁇ CRM197 ⁇ TT.23-25 Thus to find the most suitable carrier protein for preparing a vaccine these three proteins were investigated.
  • Coupling of the hapten to the carrier protein is dictated by the functional groups present on the hapten/carrier as well as solubility/aggregation compatibility. While BSA, CRM197 and TT presented no problems during hapten-protein coupling, some issues were observed in the case of KLH. Thus, at a commonly used concentration (1 mg/mL hapten and protein) significant precipitation of the conjugate occurred that is believed to be unrelated to a hapten-hydrophobicity effect that has been noted previously. Moreover, this phenomenon was exclusive to the addition of activated hapten to the protein solution rather than the hapten itself. This could be attributed to protein aggregation as KLH is a large molecule prone to form aggregates and/or precipitates.
  • the KLH was filtered through a 0.22 pm filter before adding the activated hapten material. As anticipated this process transformed the KLH to a lower aggregation state as indicated by the opalescent blue color, which persisted throughout the conjugation and purification process.
  • hapten conjugates were formulated with 500 mg of alum (alhydrogel) and 50 pg of CpG 1826 and then gently rotating at room temperature for 1 hour prior to immunizations to ensure sufficient absorption of the conjugate onto the alum surface.
  • 96-well plates (Corning 3690) were coated with 25 ng of hapten-BSA conjugate in PBS pH 7.4 (1 pg/mL, 25 pL per well) and incubated overnight at 4 °C. The plates were then washed with dH2O and blocked with 5% skim milk in PBS, pH 7.4 at room temperature for 45 minutes. Mouse serum samples were added to the first column of the plate, and a 1 : 1 serial dilution in 1% BSA-PBS, pH 7.4 was performed across each row starting with a dilution of 1 :200.
  • hapten density measurements For hapten density measurements, varying concentrations of BSA-conjugates were used to coat 96-well plates (4pg/mL, 2pg/mL, Ipg/mL, 0.5pg/mL, 0.25pg/mL, 0.125pg/mL) and the same protocol was followed vide supra. Absorbance measurements in the plateau region were plotted against the antigen coating concentrations to determine hapten density (FIGURE 9).
  • a competitive binding assay was conducted using SPR on a Biacore 3000 instrument (GE Healthcare Life Sciences) equipped with a research-grade CM5 sensor chip. Ligands were immobilized on the chip surface using standard NHSZEDC coupling chemistry with BSA in the reference flow cell (FC1) and hapten-BSA conjugate in FC2. Mouse serum was diluted in running buffer (HBS-EP+ buffer: 10 mM HEPES, 150 mM NaCl, 3 mM EDTA and 0.05% v/v Surfactant P20) and titrated on the hapten-BSA immobilized flow cell minus the reference (FC2-1) to generate a response of -100 RU following the dissociation phase.
  • HBS-EP+ buffer 10 mM HEPES, 150 mM NaCl, 3 mM EDTA and 0.05% v/v Surfactant P20
  • mice were fully anesthetized with isoflurane and rapidly decapitated using a sharp guillotine. Trunk blood was collected, and the brain was surgically separated, weighed (typically 0.4-0.5 g), and added to a tube containing zirconium oxide beads (Next Advance ZrOB05). PBS, pH 7.4 was added to the brain sample at a 1 : 1 weight ratio (i.e., 1 mL PBS for 1 g brain tissue), and the sample was homogenized using a Bullet Blender homogenizer (Next Advance). The samples were then stored at -80 °C until analysis.
  • PBS pH 7.4
  • the resulting residue was reconstituted in 100 pL of MeOH before analysis by LC-MS.
  • Drug-receptor affinity typically serves as a guidepost as to what is required for a vaccine to neutralize a drugs pharmacology.
  • the reported affinity of xylazine on the alpha2- adrenergic receptor exceeds 100 nM, which categorizes it as a weak agonist. Therefore, an immune response with affinities approximating or better than this value should readily antagonize drug-receptor binding.
  • hand-in-glove with affinity is the quantity of circulating antibody (titer) to the drug.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Mycology (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

La demande divulgue des compositions comprenant un haptène de xylazine ayant la formule (I). La demande divulgue en outre des compositions d'haptène de xylazine et des vaccins contre la xylazine. La demande divulgue en outre des procédés de prévention d'une surdose de xylazine. La demande divulgue en outre des procédés de prévention d'un trouble lié à l'utilisation de la xylazine. La demande divulgue en outre des procédés de génération d'anticorps contre la xylazine. La demande divulgue en outre des procédés de traitement de la consommation d'une substance mélangée à de la xylazine.
PCT/US2025/021994 2024-03-25 2025-03-28 Vaccin conjugué haptène contre la xylazine Pending WO2025208014A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202463569247P 2024-03-25 2024-03-25
US63/569,247 2024-03-25

Publications (1)

Publication Number Publication Date
WO2025208014A1 true WO2025208014A1 (fr) 2025-10-02

Family

ID=97218847

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2025/021994 Pending WO2025208014A1 (fr) 2024-03-25 2025-03-28 Vaccin conjugué haptène contre la xylazine

Country Status (1)

Country Link
WO (1) WO2025208014A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090068719A1 (en) * 2003-12-19 2009-03-12 Johnny Valdez Immunoassays, haptens, immunogens and antibodies for anti-hiv therapeutics
CN111454912A (zh) * 2020-04-27 2020-07-28 江南大学 一株赛拉嗪单克隆抗体杂交瘤细胞株及其应用
CN113999188A (zh) * 2021-11-10 2022-02-01 中国农业大学 赛拉嗪半抗原和人工抗原及其制备方法与应用
US20220409722A1 (en) * 2019-11-08 2022-12-29 Regents Of The University Of Minnesota Fentanyl hapten, fentanyl hapten-conjugates, and methods for making and using

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090068719A1 (en) * 2003-12-19 2009-03-12 Johnny Valdez Immunoassays, haptens, immunogens and antibodies for anti-hiv therapeutics
US20220409722A1 (en) * 2019-11-08 2022-12-29 Regents Of The University Of Minnesota Fentanyl hapten, fentanyl hapten-conjugates, and methods for making and using
CN111454912A (zh) * 2020-04-27 2020-07-28 江南大学 一株赛拉嗪单克隆抗体杂交瘤细胞株及其应用
CN113999188A (zh) * 2021-11-10 2022-02-01 中国农业大学 赛拉嗪半抗原和人工抗原及其制备方法与应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEN LINLIN, HU XIAOFEI, SUN YANING, XING YUNRUI, ZHANG GAIPING: "An ultrasensitive monoclonal antibody-based lateral flow immunoassay for the rapid detection of xylazine in milk", FOOD CHEMISTRY, vol. 383, 1 July 2022 (2022-07-01), NL , pages 132293, XP093236935, ISSN: 0308-8146, DOI: 10.1016/j.foodchem.2022.132293 *
COOPER LAURA: "Conjugate vaccine curbs xylazine effects in mice", CHEMISTRYWORLD, 31 May 2024 (2024-05-31), pages 1 - 2, XP093363078, Retrieved from the Internet <URL:https://www.chemistryworld.com/news/conjugate-vaccine-curbs-xylazine-effects-in-mice/4019569.article> DOI: 10.1039/D4CC00883A *
LIN MINGLIANG, EUBANKS LISA M., ZHOU BIN, JANDA KIM D.: "Evaluation of a hapten conjugate vaccine against the "zombie drug" xylazine", CHEMICAL COMMUNICATIONS, vol. 60, no. 35, 25 April 2024 (2024-04-25), UK , pages 4711 - 4714, XP093341156, ISSN: 1359-7345, DOI: 10.1039/D4CC00883A *

Similar Documents

Publication Publication Date Title
Raleigh et al. A fentanyl vaccine alters fentanyl distribution and protects against fentanyl-induced effects in mice and rats
CN102480959B (zh) 氢可酮的苯甲酸缀合物、苯甲酸衍生物缀合物和杂芳基羧酸缀合物及其前药、制备方法和用途
JP2002532393A (ja) エキソ−ｒ−メカミラミン製剤および治療におけるその使用
TW200418475A (en) Treatment of pain with combinations of nalbuphine and other kappa-opioid receptor agonists and opioid receptor antagonists
EA008864B1 (ru) Фармацевтические композиции для предотвращения передозировки или неправильного употребления лекарственных средств
JP6047611B2 (ja) ニコチン中毒の予防または治療用コンジュゲート
CN102573845A (zh) 新的阿片样物质的二羧酸连接的氨基酸和肽前药及其用途
WO2012122412A2 (fr) Compositions permettant de réduire le risque d&#39;effets secondaires provoqués par des interactions intermédicamenteuses
US20240009321A1 (en) Immunogenic nanovesicles for cancer immunotherapy
CN110996949A (zh) 用于治疗行为改变的方法
Hamid et al. Pre-clinical safety and toxicology profile of a candidate vaccine to treat oxycodone use disorder
Lowell et al. Vaccination against cocaine using a modifiable dendrimer nanoparticle platform
KR20220113411A (ko) KV7 칼륨 채널 개방제(opener)의 통증 치료를 위한 용도
WO2025208014A1 (fr) Vaccin conjugué haptène contre la xylazine
WO2021092446A1 (fr) Haptène de fentanyl, conjugués d&#39;haptène de fentanyl et procédés de fabrication et d&#39;utilisation de ceux-ci
Madge et al. Synthetic anti-cocaine nanoaccine successfully prevents cocaine-induced hyperlocomotion
Lin et al. Evaluation of a hapten conjugate vaccine against the “zombie drug” xylazine
US20100203084A1 (en) Method for treating pain or opioid dependence using a specific type of non-opioid agent in combination with a selective excitatory-opioid-receptor inactivator
JP2004508272A (ja) アセチルコリンエンハンサー
CN110452217B (zh) 用于尼古丁疫苗开发的光学纯半抗原
WO2024238940A1 (fr) Compositions vaccinales et leurs procédés d&#39;utilisation
US20230181728A1 (en) Adjuvanted conjugate opioid vaccine
WO2024238822A1 (fr) Compositions de vaccin à base d&#39;héroïne et procédés associés
AU2017238208A1 (en) Combination for treating pain
Moreno Immunopharmacotherapy: Towards the Creation of Effective Vaccines Against Drugs of Abuse

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: 25778552

Country of ref document: EP

Kind code of ref document: A1