Cobalt-Porphyrin Complexes for the Inactivation of the Biological Activity of Opioids

Abstract

A cobalt-loaded porphyrin complex, comprising the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) with coordinated with cobalt, effectively neutralizes the biological activity of opioids.

Claims

1. A material comprising the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) coordinated with cobalt.

2. The material of claim 1, wherein the cobalt-coordinated porphyrin is in a state of being conjugated to the surface of a nanoparticle.

3. A medicament comprising: the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) coordinated with cobalt; and a pharmaceutically-acceptable carrier.

4. The medicament of claim 3, wherein the cobalt-coordinated porphyrin is in a state of being conjugated to the surface of a nanoparticle.

5. A method of making the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) with coordinated with cobalt, comprising contacting C.sub.1S.sub.3TPP with a cobalt compound.

6. The method of claim 5, wherein the cobalt compound is cobalt chloride hexahydrate.

7. The method of claim 5, further comprising a step of conjugating the cobalt-coordinated porphyrin to the surface of a nanoparticle.

8. A method of treatment comprising: identifying a patient known or suspected to be in a state of opioid overdose, and providing the patient with a medicament including the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) coordinated with cobalt.

9. The method of claim 8, wherein the cobalt—coordinated porphyrin is in a state of being conjugated to the surface of a nanoparticle.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] FIG. 1 illustrates a cobalt-porphyrin complex (Co-TPP) and the generation thereof by loading the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) with cobalt chloride hexahydrate.

[0013] FIGS. 2A and 2B provide mass spectroscopic (MS) analysis of fentanyl incubated with Co-TPP showing the proposed mechanism for Co-C.sub.1S.sub.3TPP removal of fentanyl and mass spectrum showing putative breakdown product adducts from extratcted ion chromatogram. Co-C.sub.1S.sub.3TPP was incubated with fentanyl at 1:5 ratio for 64 hr at 37° C. FIG. 2A shows adduct formed from oxidative dealklylation of fentanyl at m/z=562.337 (arrow) and FIG. 2B shows adduct formed from simple dealkylation of fentanyl at m/z=548.332 (arrow).

[0014] FIG. 3 shows a comparative MS analysis of fentanyl incubated with metal-porphyrins complexes at a 1:1 ratio. The cobalt complex was clearly more active than the rhodium complex. Percent fentanyl remaining was determined from mass spectral abundance peak area data (counts or arbitrary units) from the extracted ion chromatogram for the accurate mass of fentanyl

[0015] FIG. 4 a comparative MS analysis of fentanyl incubated with metal-porphyrins complexes at a 5:1 ratio. The cobalt complex is clearly more active than the rhodium counterpart. Percent fentanyl remaining was determined from mass spectral abundance peak area data (counts or arbitrary units) from the extracted ion chromatogram for the accurate mass of fentanyl.

[0016] FIG. 5 plots the opioid activity of fentanyl incubated with Rh- and Co-C.sub.1S.sub.3TPP complexes. Fentanyl (0.5 mM) was incubated at equimolar concentration with Rh-C.sub.1S.sub.3TPP or Co-C.sub.1S.sub.3TPP for 24 h at 37° C. The resulting products were assayed in a mu opioid receptor activation assay. Untreated fentanyl exhibits an IC50 of 15 nM while the IC50 of fentanyl incubated with Rh-TPP or Co-C.sub.1S.sub.3TPP was 17 μm, respectively. Co-C.sub.1S.sub.3TPP shows ˜42% greater reduction in the activity of fentanyl compared to Rh-C.sub.1S.sub.3TPP.

DETAILED DESCRIPTION

Definitions

[0017] Before describing the present invention in detail, it is to be understood that the terminology used in the specification is for the purpose of describing particular embodiments, and is not necessarily intended to be limiting. Although many methods, structures and materials similar, modified, or equivalent to those described herein can be used in the practice of the present invention without undue experimentation, the preferred methods, structures and materials are described herein. In describing and claiming the present invention, the following terminology will be used in accordance with the definitions set out below.

[0018] As used herein, the singular forms “a”, “an,” and “the” do not preclude plural referents, unless the content clearly dictates otherwise.

[0019] As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items.

[0020] As used herein, the term “about” when used in conjunction with a stated numerical value or range denotes somewhat more or somewhat less than the stated value or range, to within a range of ±10% of that stated.

Overview

[0021] The invention comprises a cobalt-loaded porphyrin complex (referred to herein as “cobalt complex” or “Co-C.sub.1S.sub.3TPP complex”) that effectively neutralizes the biological activity of naturally-occurring and synthetic opioids. This occurs with greater efficacy than the rhodium-loaded porphyrin complex (“rhodium complex”) described in related U.S. Patent Application Publication No. 2020/0316085. The cobalt complex, when incubated with opioids under physiological conditions, results in the disappearance of fentanyl (used as a representative target synthetic opioid) through a heretofore unreported mechanism of the formation of fentanyl-Co breakdown adduct products. Moreover, in a tissue culture model system of opioid receptor activation, the cobalt complex inhibits fentanyl activation of the μ opioid receptor ˜42% better than the rhodium complex.

[0022] It is expected that delivery of the cobalt complex to a patient known or suspected of suffering an opioid overdose might be effective to ameliorate the effects of the overdose. Thus, a medicament is contemplated comprising the cobalt complex in conjunction with a pharmaceutically-acceptable carrier.

Examples

[0023] The Co-TPP complex was prepared by loading the porphyrin (meso-tri(4-sulfonatophenyl) mono(4-carboxyphenyl)porphine (C.sub.1S.sub.3TPP)) with cobalt chloride hexahydrate generally following a procedure previously described in the literature which involved a different porphyrin, X. Fu and B.B. Wayland, J. Am. Chem. Soc., 2004, 126, 2623, incorporated herein by reference for disclosing a technique for preparing a metal/porphyrin complex. As indicated in FIG. 1, incubation in ethanol/water for 30 minutes resulted in formation of the complex.

[0024] The interaction of the Co-C.sub.1S.sub.3TPP complex with fentanyl was first characterized using mass spectroscopy (MS) to determine the speciation of the products generated after Co-TPP was incubated with fentanyl (at 1:5 ratio) after 64 h at 37° C. FIGS. 2A and 2B show the results of MS analysis after fentanyl was incubated with the Co-C.sub.1S.sub.3TPP complex (at 1:5 ratio) for 64 h at 37° C. The data show the disappearance of fentanyl (as tracked by quantifying the molecular weight peak of fentanyl) and the appearance of a fentanyl: Co-C.sub.1S.sub.3TPP adduct (m/z=562.337) formed from oxidative dealkylation of fentanyl (FIG. 2B). FIG. 2b shows the formation of a fentanyl: Co-C.sub.1S.sub.3TPP adduct (m/z=548.332) formed from simple dealkylation of fentanyl. The formation of either adduct product results in the disappearance of the fentanyl molecular weight peak in this analysis.

[0025] Co-C.sub.1S.sub.3TPP was incubated for 72 h at 37° C. to ascertain a time-resolved comparison of the disappearance of fentanyl mediated by either the Rh—or Co-C.sub.1S.sub.3TPP complex. FIG. 3 shows the results when the complexes were incubated at a 1:1 ratio. The Co-C.sub.1S.sub.3TPP complex clearly mediated a faster disappearance of fentanyl than did Rh-TPP over the 72 h time course. This effect was even further enhanced when the ratio of complex to fentanyl was increased to 5:1 (FIG. 4).

[0026] FIG. 5 shows data from a classical cell-based opioid cellular signaling assay used to ascertain the biological activity of fentanyl after its incubation with the Rh—or Co-Co-C.sub.1S.sub.3TPP complexes. Chinese hamster ovary cells that stably express the mu opioid receptor were incubated with fentanyl alone or fentanyl that was treated with equimolar Rh-C.sub.1S.sub.3TPP or Co-C.sub.1S.sub.3TPP for 24 h. The ability of fentanyl to inhibit the formation of cyclic AMP is determined by measuring the cellular levels of free ATP. Here, an increase in photoluminescence intensity quantitatively tracks active fentanyl in a dose dependent manner. While Rh-C.sub.1S.sub.3TPP reduced the opioid activity of fentanyl˜113—fold from an IC50 of 15 nM to 1.7 μM, Co-C.sub.1S.sub.3TPP reduced fentanyl activity ˜160-fold to 2.4 μM. This corresponds to a 42% increase in the opioid-neutralizing activity fentanyl for the Co-TPP complex compared to the Rh-C1S3TPP complex.

Further Embodiments

[0027] It is expected that other metals besides could be used as a substitute in the porphyrin complex resulting in tailored activity.

[0028] The cobalt-porphyrin complex can be conjugated to and displayed on the surface of, or in the core of, various nanoparticles. It is expected that activity of the Co-TPP complex might be augmented in this fashion, just as was activity of the rhodium complex described in related U.S. Patent Application Publication No. 2020/0316085. Examples of these nanoparticles include, but are not limited to liposomes, gold nanoparticles, metal oxide particles, quantum dots, polymers, nucleic acids.

[0029] Other porphyrins could be used to generate the opioid-neutralizing complex.

[0030] Pharmaceutically acceptable carriers include carriers that do not themselves induce the production of antibodies harmful to the individual receiving the composition. Suitable carriers are typically large, slowly metabolized macromolecules such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, and inactive virus particles. The carrier can comprise, consist of, consist essentially of, or be a saline solution, dextrose, albumin, a serum, or any of those disclosed in U.S. Pub. Nos.: 2008/0138408; 2009/0061003; 2009/0123530; 2010/0303901; 2012/0034198; and 2016/0008290 and U.S. Pat. Nos.: 6,992,066; 5,785,973; 7,485,294; 8,088,734; 8,753,645; 8,808,733; and 8,858,998.

[0031] The compositions typically will contain pharmaceutically acceptable vehicles, such as water, saline, glycerol, ethanol, etc. Additionally, auxiliary substances, such as wetting or emulsifying agents, pH buffering substances, preservatives, and the like, may be included in such vehicles.

[0032] Typically, the compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation also may be emulsified or encapsulated in liposomes. Solutions for infusion or injection may be prepared in a conventional manner, e.g. with the addition of preservatives such as p-hydroxybenzoates or stabilizers such as alkali metal salts of ethylenediamine tetraacetic acid, which may then be transferred into fusion vessels, injection vials or ampules. Alternatively, the compound for injection may be lyophilized either with or without the other ingredients and be solubilized in a buffered solution or distilled water, as appropriate, at the time of use. Non-aqueous vehicles such as fixed oils and ethyl oleate are also useful herein.

[0033] In cases where intramuscular injection is the mode of administration, an isotonic formulation can be used. Generally, additives for isotonicity can include sodium chloride, dextrose, mannitol, sorbitol and lactose. In some cases isotonic solutions such as phosphate buffered saline are preferred. Stabilizers include glycerol, gelatin and albumin which may be included in the formulation. In some embodiments, a vasoconstriction agent is added to the formulation.

[0034] Additional pharmaceutical methods may be employed to control the duration of action. Controlled release preparations may be achieved through the use of polymers to complex or absorb the compounds. The controlled delivery may be exercised by selecting appropriate macromolecules (for example polyesters, polyamino acids, polyvinyl, pyrrolidone, ethylenevinylacetate, methylcellulose, carboxymethylcellulose, or protamine sulfate) and the concentration of macromolecules as well as the method of incorporation in order to control release. Another possible method to control the duration of action by controlled release preparations is to incorporate the compounds of the present invention into particles of a polymeric material such as polyesters, polyamino acids, hydrogels, polylactic acid or ethylene vinylacetate copolymers. Alternatively, instead of incorporating these agents into polymeric particles, it is possible to entrap these materials in microcapsules prepared, for example, interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly(methylmethacylate)-microcapsules, respectively, or in colloidal drug delivery systems, for example, liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules or in macroemulsions.

Concluding Remarks

[0035] Although the present invention has been described in connection with preferred embodiments thereof, it will be appreciated by those skilled in the art that additions, deletions, modifications, and substitutions not specifically described may be made without departing from the spirit and scope of the invention. Terminology used herein should not be construed as being “means-plus-function” language unless the term “means” is expressly used in association therewith.

REFERENCES

[0036] [1] H. Hedegaard et al. (2018) “Drugs Most Frequently Involved in Drug Overdose Deaths: United States, 2011-2016.” National Vital Statistics Reports 67: 1-13.

[0037] [2] J.R. Riches et al. (2012) “Analysis of Clothing and Urine from Moscow Theatre Siege Casualties Reveals Carfentanil and Remifentanil Use” Journal of Analytical Toxicology 36:647-656.