COMPOSITIONS AND METHODS FOR THE PREVENTION AND TREATMENT OF ERGOT ALKALOID TOXICITY

Abstract

Embodiments of the present invention provide materials and methods for preventing and treating ergot-based toxicity in animals, including humans. In particular, the present disclosure provides materials and methods for ameliorating the harmful physical manifestations of various diseases caused, at least in part, by ergot-based toxicity, including but not limited to caudal heel pain syndrome, idiopathic headshaking, pituitary pars intermedia dysfunction, metabolic syndrome and laminitis in horses; fescue foot, infertility and summer slump in cattle, sheep and goats; and neurologic, mental and somatic disorders in humans.

Claims

1. A composition for treating ergot-based toxicity in a subject, the composition comprising one or more compounds selected from the group consisting of: ##STR00022## wherein R.sup.1 is selected from the group consisting of hydrogen and null, in the case of a double-bond between the C-3 and C-4 carbons; R.sup.2 is selected from the group consisting of a methyl and hydrogen; X is selected from the group consisting of a bond, carbon, nitrogen, oxygen, an amine, an amide, an ester, and an ether; Y is selected from the group consisting of hydrogen, methyl, ethyl, ##STR00023## Z is selected from oxygen and nitrogen; R.sup.3 is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, sec-butyl; R.sup.4 is selected from the group consisting of benzyl, ethyl, isopropyl, isobutyl, sec-butyl, n-butyl, 2-methyl-n-butyl, 2-methyl-n-propyl, and ethyl(methyl)sulfane; R.sup.5 is selected from the group consisting of hydrogen and methoxy; R.sup.6 is selected from the group consisting of isopropyl and sec-butyl; and R.sup.7 is selected from the group consisting of benzyl, ethyl, isopropyl, isobutyl, and sec-butyl.

2. The composition of claim 1, wherein the carrier molecule is selected from the group consisting of human serum albumin, bovine serum albumin, chicken globulin, ovalbumin, keyhole limpet hemocyanin, polyarginine, polyhistidine, polytyrosine, polyserine, polyaspartate, and polylysine.

3. The composition of claim 1, wherein at least one of the compounds has the following structure: ##STR00024##

4. The composition of claim 1, wherein at least one of the compounds has the following structure: ##STR00025##

5. The composition of claim 1, wherein at least one of the compounds has the following structure: ##STR00026##

6. The composition of claim 1, wherein at least one of the compounds has the following structure: ##STR00027##

7. The composition of claim 1, wherein at least one of the compounds has hydrogen at R.sup.1.

8. The composition of claim 1, wherein at least one of the compounds is selected from the group consisting of ##STR00028## ##STR00029## ##STR00030## ##STR00031##

9. A composition for treating ergot-based toxicity in a subject, the composition comprising one or more clavines bonded to a carrier molecule.

10. The composition of claim 9, wherein the composition is selected from at least one of the group consisting of: ##STR00032## ##STR00033## ##STR00034## ##STR00035## R.sup.8 is selected from the group consisting of hydrogen and hydroxyl; R.sup.9 is selected from the group consisting of or hydrogen, and or hydroxyl; R.sup.10 is selected from the group consisting of hydrogen, hydroxyl, ##STR00036## R.sup.11 is selected from the group consisting of and hydrogen; R.sup.12 is selected from the group consisting of or hydrogen, or hydroxyl, and or acetoxy; R.sup.13 is selected from the group consisting of and hydrogen; R.sup.14 is selected from the group consisting of hydrogen and ##STR00037## R.sup.15 is selected from the group consisting of methyl, CH.sub.2OH, COH, ##STR00038## R.sup.16 is selected from the group consisting of methyl, CH.sub.2OH, and hydroxyl; R.sup.17 is selected from the group consisting of hydrogen and methyl; R.sup.18 is selected from the group consisting of hydrogen and methoxy; R.sup.19 is selected from the group consisting of hydrogen and chloride; R.sup.20 is selected from the group consisting of and NHCH.sub.3; R.sup.21 is selected from the group consisting of and ##STR00039## and R.sup.22 is selected from the group consisting of and COOH.

11. A composition for preventing ergot toxicity, wherein at least one compound in the composition is: ##STR00040## wherein Z is selected from oxygen and nitrogen.

12. A method for treating a subject exhibiting one or more physical manifestations of ergot-based toxicity, the method comprising administering a composition of claim 1.

13. A method for treating a subject exhibiting one or more physical manifestations of ergot-based toxicity, the method comprising administering a composition of claim 10.

Description

DETAILED DESCRIPTION

[0020] Embodiments of the present invention provide materials and methods for treating ergot-based toxicity in animals. In particular, the present disclosure provides materials and methods for ameliorating the harmful physical manifestations of various diseases due to ergot-based toxicity, at least in part. Important diseases to prevent include summer slump and fescue foot in cattle along with caudal heel pain syndrome, idiopathic headshaking, and laminitis in horses.

[0021] The present disclosure addresses the need for therapeutic methods and treatments to reduce the harmful effects of ergot-based toxicity. For example, in some embodiments, the methods and treatments of the present disclosure can mitigate physical manifestations of ergot toxicity, such as preventing vasoconstriction in the extremities, reducing infertility and increasing average daily gains of animals. In some embodiments, preventing vasoconstriction will ameliorate one or more symptoms associated with diseases like caudal heel pain syndrome, idiopathic headshaking syndrome, and laminitis, for example.

[0022] Many animal species may be impacted by the presence of ergot derivatives in their diet, including, but not limited to mammals, such as horses, cows, pigs, sheep, goats, dogs, cats, humans and so on. While vaccination against ergot derivatives has been attempted (See, e.g., Fillipov et al., J. Anim. Sci., 1998, 76(9), 2456-2463; those authors found a lack of long-lasting protection), there remains a profound need for an answer to the debilitating impact of ergot derivatives in animals. The present invention provides vaccines based on a unique chemical design.

[0023] Embodiments of the present invention are included to demonstrate certain embodiments presented herein. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered to function well in the practices disclosed herein.

[0024] This disclosure provides various immunogenic compounds and compositions described below. In general, many different forms of vaccine delivery are available to one of ordinary skill in the art. Descriptions of such methods are widely available in the art. One article which reviews such methods is Saroja, P. K., et al., Int. J Pharm. Invest., 2011, 1.2, 64-74. Additional discussion about vaccine delivery for animals may be found at Sharma S, Hinds LA. Formulation and delivery of vaccines: Ongoing challenges for animal management. Journal of Pharmacy & Bioallied Sciences 2012; 4(4), 258-266. The compounds of the invention are suitable for oral or mucosal delivery. However, the fructofuranosides or fructofuranosyl fructofuranosides may be more difficult to administer orally due to acid lability.

[0025] The compounds of the invention may be used as desired to vaccinate against any number of ergot-based toxicities. The user may choose which of the compounds are desired to be vaccinated against. It is not required that all compounds of the invention be used, only those for which protection is desired. The compounds of the invention may be in a sustained release preparation. These preparations may be utilized as a prophylactic to protect animals, such as mammals (e.g., humans, horses, etc.) from the toxic effects of ergopeptine and clavine alkaloids. Alternatively, the various preparations can be used as can be used as a therapeutic. As can be appreciated by one skilled in the art, there are many suitable ways to incorporate the immunogenic compounds described below into various embodiments of sustained release preparations (e.g., microcapsules, microsphere polymers, liposomes, polylactic acid preparations, etc.). Various embodiments disclosed herein may utilize the various immunogenic compounds described below in a biocompatible, biodegradable microsphere polymer or copolymer of polylactide or polyglycolide.

[0026] In various embodiments, the antigen can be incorporated, for example, into biodegradable microspheres, to produce an immunizing agent that will result in prolonged release of the antigen and therefore induce a long term immune response. Exemplary agents to make vaccine-containing microspheres include polyesters of polylactic acid, polyglycolic acid, their respective co-polymers, and combinations thereof. Exemplary microspheres may be produced using mild conditions that do not degrade or damage the various antigens. The antigens may be enclosed in the biodegradable matrix. Three exemplary methods used to produce these microspheres include phase separation (e.g., where drugs and polymers are dispersed or dissolved in a solvent and then the microspheres may be precipitated out by addition of silicon oil), solvent extraction (e.g., where drugs and polymers in solution are added to an aqueous solution of poly-vinyl alcohol to produce an oil-in-water emulsion and then the solvent is eliminated by adding water and the microspheres dried), and spray drying (e.g., where drugs and polymers are dissolved in a solvent and then spray dried). In any of the aforementioned procedures, after the spheres are formed, they may be dried and then separated into various sizes, for example, by sieving.

[0027] Factors which affect antigen release include erosion and breakdown of the particles, diffusion of the drug out of the matrix, solubility of the antigen, antigen molecular weight, antigen loading of the spheres and polymer molecular weight. For a given antigen the release rate is related to particle size; small particles release the antigen sooner than large particles. For prolonged release and immunization a mixture of small and large particles appears to be desirable as would be appreciated by an ordinary skilled artisan having the benefit of this disclosure.

[0028] The immunogenic compounds, described in further detail below, whether or not contained in a biodegradable microsphere, may also be placed in a pharmaceutically acceptable carrier, including but not limited to buffered saline or distilled water. Likewise, the immunogenic compounds can be mixed with a suitable adjuvant.

[0029] Chemical synthesis of the ergot derivatives can be achieved via methods known in the art. See, e.g., Recent Synthetic Studies on the Ergot Alkaloids and Related Compounds. The Alkaloids: Chemistry and Biology, Academic Press: San Diego, Calif., 2000; Vol. 54, pp 191-257. See also Liu and Jia, Nat. Prod. Rep., 2017, 34, 411-432.

[0030] In the aforementioned disclosed compounds, the carrier molecule may include, but is not limited to, a peptide or a protein. Exemplary proteins include a suitable immunogenic protein, which may include human serum albumin, bovine serum albumin, chicken globulin, ovalbumin, keyhole limpet hemocyanin, tetanus toxoid, polyarginine, polyhistidine, polytyrosine, polyserine, polyaspartate, and polylysine. A review of such molecules can be found at Pichichero, Michael E. Hum. Vaccin. Immunother., 2013, 9(12), 2505-2523. Methods for attaching such molecules are also well-known in the art.

[0031] In general, methods for synthesis of the subject compounds take advantage of the ability to deprotonate the indole NH using strong bases such as NaH in a suitable solvent such as dioxane, DMSO, and other solvents known to those skilled in the art. The resulting indolic anion is then treated with, for example, methyl-4-bromobutyrate to generate the 1-(4-carbomethoxypropyl)indole derivative. This methyl ester is then selectively hydrolyzed by mild base or lithium iodide to afford the free carboxylic acid. The resulting 1-(3-carboxypropyl)indole derivative is then coupled to the carrier molecule, usually a protein of interest, using standard peptide coupling reagents such as carbonyl diimidazole, a carbodiimide reagent, or the like. Following coupling the protein thus modified is purified to remove any excess uncoupled ergot derivative and byproducts of the coupling reagent, as shown below.

##STR00001##

[0032] The ergot alkaloid derivatives themselves can be prepared from commercially available lysergic acid using known methods (U.S. Pat. No. 3,336,311 and Liu, H. et al., Org. Lett., 2017, 19(12), 3323-3326), while the clavines can also be prepared in accordance with known methods (see: Ergot Alkaloids. The Alkaloids: Chemistry and Pharmacology, Academic Press: San Diego, Calif., 1990; Vol. 38, pp 1-156; Liu, H. et al., Org. Lett., 2017, 19(12), 3323-3326; Oppolzer et al., Tetrahedron, 1983, 39(22), 3695-3705; McCabe, S. and Wipf, P., Org. and Biomol. Chem., 2016, 14, 5894-5913; McCamley, K. et al., J. Org. Chem., 2003, 68(25), 9847-9850; Schkeryantz J., et al., J. Am. Chem. Soc., 1999, 121, 11964-11975; Ken, V. et al., Appl. Microbiol. Biotech., 1990, 32, 645-650, Peng, Y. and Li, W.-D. Synlett, 2006, 1165-1168, Liu, Z., et al. J. Org. Chem., 2014, 79, 11792-11796).

In general, those molecules with free hydroxyl groups must be protected prior to the addition of methyl-4-bromobutyrate. An example is shown below.

##STR00002##

[0033] In situations in which a free amine is present, protection of the amine would also be required. A further example is shown below.

##STR00003##

[0034] Finally, in those cases in which fructofuranosides or fructofuranosyl fructofuranosides are desired, the following reaction sequence may be employed to synthesize the compounds, as shown with the exemplary compound below.

##STR00004##

[0035] A composition for treating ergot-based toxicity in a subject is a compound of Formula 1:

##STR00005##

[0036] R.sup.1 is selected from the group consisting of hydrogen and null in the case of the indicated double bond. Thus, the composition of Formula 1 includes one or more of the following structures:

##STR00006##

[0037] wherein R.sup.2 is selected from the group consisting of methyl and hydrogen;

[0038] Z is selected from the group consisting of oxygen and nitrogen;

Y is selected from the group consisting of hydrogen, methyl, ethyl,

##STR00007##

R.sup.3 is selected from the group consisting of methyl, ethyl, n-propyl, isopropyl, and sec-butyl;
R.sup.4 is selected from the group consisting of benzyl, ethyl, isopropyl, isobutyl, sec-butyl, n-butyl, 2-methyl-n-butyl, 2-methyl-n-propyl, and ethyl(methyl)sulfane;
R.sup.5 is selected from the group consisting of hydrogen and methoxy;
R.sup.6 is selected from the group consisting of isopropyl and sec-butyl;
R.sup.7 is selected from the group consisting of benzyl, ethyl, isopropyl, isobutyl, and sec-butyl; and

[0039] X is selected from the group consisting of a bond, carbon, nitrogen, oxygen, an amine (e.g., a primary amine, a secondary amine, or a tertiary amine), an amide (e.g., a primary amide, a secondary amide, or a tertiary amide), an ester, and an ether.

[0040] Exemplary compounds of Formula 1 include:

##STR00008## ##STR00009## ##STR00010## ##STR00011##

[0041] Also disclosed herein are compositions for treating ergot-based toxicity in a subject where the composition includes a clavine bonded to a carrier molecule. Exemplary clavines include the following compounds:

##STR00012## ##STR00013##

[0042] wherein R.sup.8 is selected from the group consisting of hydrogen and hydroxyl;

[0043] R.sup.9 is selected from the group consisting of or hydrogen, and or hydroxyl;

[0044] R.sup.10 is selected from the group consisting of hydrogen, hydroxyl,

##STR00014##

[0045] R.sup.11 is selected from the group consisting of and hydrogen;

[0046] R.sup.12 is selected from the group consisting of or hydrogen, or hydroxyl, and or acetoxy;

[0047] R.sup.13 is selected from the group consisting of and hydrogen;

[0048] R.sup.14 is selected from the group consisting of hydrogen and

##STR00015##

[0049] R.sup.15 is selected from the group consisting of methyl, CH.sub.2OH, COH,

##STR00016##

[0050] R.sup.16 is selected from the group consisting of methyl, CH.sub.2OH, and hydroxyl;

[0051] R.sup.17 is selected from the group consisting of hydrogen and methyl;

[0052] R.sup.18 is selected from the group consisting of hydrogen and methoxy;

[0053] R.sup.19 is selected from the group consisting of hydrogen and chloride;

[0054] R.sup.20 is selected from the group consisting of and NHCH.sub.3;

[0055] R.sup.21 is selected from the group consisting of and

##STR00017##

and

[0056] R.sup.22 is selected from the group consisting of and COOH.

[0057] Thus, in various embodiments, compounds of the present invention include:

##STR00018## ##STR00019## ##STR00020## ##STR00021##

[0058] In the aforementioned disclosed compounds, the carrier molecule is not particularly limited and may include, but is not limited to, a peptide or a protein. Exemplary proteins include a suitable immunogenic protein, which may include human serum albumin, bovine serum albumin, chicken globulin, ovalbumin, keyhole limpet hemocyanin, polyarginine, polyhistidine, polytyrosine, polyserine, polyaspartate, and polylysine.

[0059] Also disclosed herein are methods for treatment of a subject or animal, such as a mammal exhibiting one or more physical manifestations of ergot-based toxicity. Various methods include administering a therapeutic or immunogenic amount of one or more of the aforementioned compounds and treating the one or more physical manifestation of ergot-based toxicity in the subject.

[0060] All of the MATERIALS and METHODS disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods have been described in terms of preferred embodiments, it is apparent to those of skill in the art that variations maybe applied to the MATERIALS and METHODS and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope herein. More specifically, certain agents that are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept as defined by the appended claims.