Drug Design and Synthesis Section – Publications

@article{pmid35268659,

title = {Design, Synthesis, and In Vivo Evaluation of C1-Linked 4,5-Epoxymorphinan Haptens for Heroin Vaccines},

author = {Agnieszka Sulima and Fuying Li and Jeffrey Brian Morgan and Phong Truong and Joshua F G Antoline and Therese Oertel and Rodell C Barrientos and Oscar B Torres and Zoltan Beck and Gregory H Imler and Jeffrey R Deschamps and Gary R Matyas and Arthur E Jacobson and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/35268659/},

doi = {10.3390/molecules27051553},

issn = {1420-3049},

year  = {2022},

date = {2022-02-01},

urldate = {2022-02-01},

journal = {Molecules},

volume = {27},

number = {5},

abstract = {In our continuing effort to develop effective anti-heroin vaccines as potential medications for the treatment of opioid use disorder, herein we present the design and synthesis of the haptens: 1-AmidoMorHap (), 1-AmidoMorHap epimer (), 1 Amido-DihydroMorHap (), and 1 Amido-DihydroMorHap epimer (). This is the first report of hydrolytically stable haptenic surrogates of heroin with the attachment site at the C1 position in the 4,5-epoxymorophinan nucleus. We prepared respective tetanus toxoid (TT)-hapten conjugates as heroin vaccine immunogens and evaluated their efficacy in vivo. We showed that all TT-hapten conjugates induced high antibody endpoint titers against the targets but only haptens  and  can induce protective effects against heroin in vivo. The epimeric analogues of these haptens,  and , failed to protect mice from the effects of heroin. We also showed that the in vivo efficacy is consistent with the results of the in vitro drug sequestration assay. Attachment of the linker at the C1 position induced antibodies with weak binding to the target drugs. Only TT- and TT- yielded antibodies that bound heroin and 6-acetyl morphine. None of the TT-hapten conjugates induced antibodies that cross-reacted with morphine, methadone, naloxone, or naltrexone, and only TT- interacted weakly with buprenorphine, and that subtle structural difference, especially at the C6 position, can vastly alter the specificity of the induced antibodies. This study is an important contribution in the field of vaccine development against small-molecule targets, providing proof that the chirality at C6 in these epoxymorphinans is a vital key to their effectiveness.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34693882,

title = {Novel chimeric monoclonal antibodies that block fentanyl effects and alter fentanyl biodistribution in mice},

author = {Bhupal Ban and Rodell C Barrientos and Therese Oertel and Essie Komla and Connor Whalen and Megan Sopko and Yingjian You and Partha Banerjee and Agnieszka Sulima and Arthur E Jacobson and Kenner C Rice and Gary R Matyas and Vidadi Yusibov},

url = {https://pubmed.ncbi.nlm.nih.gov/34693882/},

doi = {10.1080/19420862.2021.1991552},

issn = {1942-0870},

year  = {2021},

date = {2021-12-01},

journal = {MAbs},

volume = {13},

number = {1},

pages = {1991552},

abstract = {The prevalence and societal impact of opioid use disorder (OUD) is an acknowledged public health crisis that is further aggravated by the current pandemic. One of the devastating consequences of OUD is opioid overdose deaths. While multiple medications are now available to treat OUD, given the prevalence and societal burden, additional well-tolerated and effective therapies are still needed. To this point, we have developed chimeric monoclonal antibodies (mAb) that will specifically complex with fentanyl and its analogs in the periphery, thereby preventing them from reaching the central nervous system. Additionally, mAb-based passive immunotherapy offers a high degree of specificity to drugs of abuse and does not interfere with an individual's ability to use any of the medications used to treat OUD. We hypothesized that sequestering fentanyl and its analogs in the periphery will mitigate their negative effects on the brain and peripheral organs. This study is the first report of chimeric mAb against fentanyl and its analogs. We have discovered, engineered the chimeric versions, and identified the selectivity of these antibodies, through  characterization and  animal challenge studies. Two mAb candidates with very high (0.1-1.3 nM) binding affinities to fentanyl and its analogs were found to be effective in engaging fentanyl in the periphery and blocking its effects in challenged animals. Results presented in this work constitute a major contribution in the field of novel therapeutics targeting OUD.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34205869,

title = {Effect of Preexisting Immunity to Tetanus Toxoid on the Efficacy of Tetanus Toxoid-Conjugated Heroin Vaccine in Mice},

author = {Essie Komla and Oscar B Torres and Rashmi Jalah and Agnieszka Sulima and Zoltan Beck and Carl R Alving and Arthur E Jacobson and Kenner C Rice and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/34205869/},

doi = {10.3390/vaccines9060573},

issn = {2076-393X},

year  = {2021},

date = {2021-06-01},

urldate = {2021-06-01},

journal = {Vaccines (Basel)},

volume = {9},

number = {6},

abstract = {Opioid use disorder (OUD) is a serious health problem that has dramatically increased over the last decade. Although current therapies for the management of OUD can be effective, they have limitations. The complementary strategy to combat the opioid crisis is the development of a conjugate vaccine to generate high affinity antibodies in order to neutralize opioids in circulation before reaching the brain. The components of an opioid vaccine include an opioid hapten (6-AmHap) that is conjugated to a carrier protein (tetanus toxoid) with the addition of adjuvants (Army Liposome Formulation adsorbed to aluminum hydroxide-ALFA). There is no consensus in the literature as to whether preexisting immunity to the carrier protein may impact the immunogenicity of the conjugate vaccine by inducing an enhanced or suppressed immune response to the hapten. Here, we investigated whether pre-exposure to tetanus toxoid would affect the immunogenicity and efficacy of the heroin vaccine, TT-6-AmHap. Mice were primed with diphtheria, tetanus, and acellular pertussis (DTaP) vaccine at weeks -4 and -2, then immunized with TT-6-AmHap vaccine at weeks 0, 3, and 6. Using ELISA and behavioral assays, we found that preexisting immunity to tetanus toxoid had no influence on the immunogenicity and efficacy of the TT-6-AmHap vaccine.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34179783,

title = {Synthesis and immunological effects of C14-linked 4,5-epoxymorphinan analogues as novel heroin vaccine haptens},

author = {Eugene S Gutman and Thomas C Irvin and J Brian Morgan and Rodell C Barrientos and Oscar B Torres and Zoltan Beck and Gary R Matyas and Arthur E Jacobson and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/34179783/},

doi = {10.1039/d1cb00029b},

issn = {2633-0679},

year  = {2021},

date = {2021-04-01},

urldate = {2021-04-01},

journal = {RSC Chem Biol},

volume = {2},

number = {3},

pages = {835--842},

abstract = {Active immunization is being explored as a potential therapeutic to combat accidental overdose and to mitigate the abuse potential of opioids. Hapten design is one of the crucial factors that determines the efficacy of a candidate vaccine to substance abuse and remains one of the most active areas of research in vaccine development. Herein we report for the first time the synthesis of three novel opiate surrogates with the linker attachment site at C14,  (6,14-AmidoHap),  (14-AmidoMorHap), and  (14-AmidoHerHap) as novel heroin haptens. The compounds , , and  are analogues with different substituents at C6: an acetamide, a hydroxyl moiety, and an acetate, respectively. All three haptens had a phenolic hydroxyl group at C3. The haptens were conjugated to the tetanus toxoid carrier protein, adjuvanted with liposomal monophosphoryl lipid A/aluminum hydroxide and were tested in mice in terms of immunogenicity and efficacy. Immunization of mice resulted in antibody endpoint titers of >10 against all the haptens. Neither of the conjugates of , , and  had induced antibodies with selectivity broad enough to recognize and bind heroin, 6-AM, and morphine resulting in little to no protection against the antinociceptive effects of heroin . Only the mice immunized with conjugate  were partially protected against heroin-induced antinociception. These results contribute to the growing body of knowledge that the linker position and the subtle structural differences in the hapten scaffold impact the selectivity of the induced antibodies. Together, these highlight the importance of rational hapten design for heroin vaccine development.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34076427,

title = {Bivalent Conjugate Vaccine Induces Dual Immunogenic Response That Attenuates Heroin and Fentanyl Effects in Mice},

author = {Rodell C Barrientos and Connor Whalen and Oscar B Torres and Agnieszka Sulima and Eric W Bow and Essie Komla and Zoltan Beck and Arthur E Jacobson and Kenner C Rice and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/34076427/},

doi = {10.1021/acs.bioconjchem.1c00179},

issn = {1520-4812},

year  = {2021},

date = {2021-01-01},

urldate = {2021-01-01},

journal = {Bioconjug Chem},

volume = {32},

number = {11},

pages = {2295--2306},

abstract = {Opioid use disorders and fatal overdose due to consumption of fentanyl-laced heroin remain a major public health menace in the United States. Vaccination may serve as a promising potential remedy to combat accidental overdose and to mitigate the abuse potential of opioids. We previously reported the heroin and fentanyl monovalent vaccines carrying, respectively, a heroin hapten, 6-AmHap, and a fentanyl hapten, AmFenHap, conjugated to tetanus toxoid (TT). Herein, we describe the mixing of these antigens to formulate a bivalent vaccine adjuvanted with liposomes containing monophosphoryl lipid A (MPLA) adsorbed on aluminum hydroxide. Immunization of mice with the bivalent vaccine resulted in IgG titers of >10 against both haptens. The polyclonal sera bound heroin, 6-acetylmorphine, morphine, and fentanyl with dissociation constants () of 0.25 to 0.50 nM. Mice were protected from the anti-nociceptive effects of heroin, fentanyl, and heroin +9% (w/w) fentanyl. No cross-reactivity to methadone and buprenorphine was observed . Naloxone remained efficacious in immunized mice. These results highlighted the potential of combining TT-6-AmHap and TT-AmFenHap to yield an efficacious bivalent vaccine that could ablate heroin and fentanyl effects. This vaccine warrants further testing to establish its potential translatability to humans.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34011153b,

title = {Novel Dual-Target μ-Opioid Receptor and Dopamine D Receptor Ligands as Potential Nonaddictive Pharmacotherapeutics for Pain Management},

author = {Alessandro Bonifazi and Francisco O Battiti and Julie Sanchez and Saheem A Zaidi and Eric Bow and Mariia Makarova and Jianjing Cao and Anver Basha Shaik and Agnieszka Sulima and Kenner C Rice and Vsevolod Katritch and Meritxell Canals and J Robert Lane and Amy Hauck Newman},

url = {https://pubmed.ncbi.nlm.nih.gov/34011153/},

doi = {10.1021/acs.jmedchem.1c00611},

issn = {1520-4804},

year  = {2021},

date = {2021-01-01},

urldate = {2021-01-01},

journal = {J Med Chem},

volume = {64},

number = {11},

pages = {7778--7808},

abstract = {The need for safer pain-management therapies with decreased abuse liability inspired a novel drug design that retains μ-opioid receptor (MOR)-mediated analgesia, while minimizing addictive liability. We recently demonstrated that targeting the dopamine D receptor (DR) with highly selective antagonists/partial agonists can reduce opioid self-administration and reinstatement to drug seeking in rodent models without diminishing antinociceptive effects. The identification of the DR as a target for the treatment of opioid use disorders prompted the idea of generating a class of ligands presenting bitopic or bivalent structures, allowing the dual-target binding of the MOR and DR. Structure-activity relationship studies using computationally aided drug design and  binding assays led to the identification of potent dual-target leads (, , and ), based on different structural templates and scaffolds, with moderate (sub-micromolar) to high (low nanomolar/sub-nanomolar) binding affinities. Bioluminescence resonance energy transfer-based functional studies revealed MOR agonist-DR antagonist/partial agonist efficacies that suggest potential for maintaining analgesia with reduced opioid-abuse liability.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid34662828,

title = {Synthesis of PF-6870961, a major hydroxy metabolite of the novel ghrelin receptor inverse agonist PF-5190457},

author = {Agnieszka Sulima and Fatemeh Akhlaghi and Lorenzo Leggio and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/34662828/},

doi = {10.1016/j.bmc.2021.116465},

issn = {1464-3391},

year  = {2021},

date = {2021-01-01},

urldate = {2021-01-01},

journal = {Bioorg Med Chem},

volume = {50},

pages = {116465},

abstract = {Preclinical and human studies have indicated involvement of the ghrelin system in alcohol-related behaviors illuminating the possibility of using ghrelin receptor blockers as a pharmacological intervention for alcohol use disorder (AUD). Preliminary data from a recently conducted phase 1b human study with a ghrelin receptor inverse agonist, PF-5190457 (2-(2-methylimidazo[2,1-b][1,3thiazol-6-yl)-1-{2-(1R)-5-(6-methylpyrimidin-4-yl)-2,3-dihydro-1H-inden-1-yl]-2,7-diazaspiro[3.5]non-7-ylethanone), provided evidence on the safety and tolerability of this compound when co-administered with alcohol. Furthermore, the study revealed important information on the biotransformation pathways for this compound and prompted the discovery and then synthesis of a newly identified major metabolite, PF-6870961 ((R)-1-(2-(5-(2-hydroxy-6-methylpyrimidin-4-yl)-2,3-dihydro-1H-inden-1-yl)-2,7-diazaspiro[3.5]nonan-7-yl)-2-(2-methylimidazo[2,1-b]thiazol-6-yl)ethan-1-one). The metabolite was synthesized and fully characterized through a design that enabled it to be prepared in useful quantities. The synthesis provided direct access to the recently discovered PF-6870961 and is allowing researchers to conduct additional and deeper evaluation of its in vitro and in vivo properties.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid33479684,

title = {G-Protein biased opioid agonists: 3-hydroxy--phenethyl-5-phenylmorphans with three-carbon chain substituents at C9},

author = {Eugene S Gutman and Eric Bow and Fuying Li and Agnieszka Sulima and Sophia Kaska and Rachel Crowley and Thomas E Prisinzano and Yong-Sok Lee and Sergio A Hassan and Gregory H Imler and Jeffrey R Deschamps and Arthur E Jacobson and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/33479684/},

doi = {10.1039/d0md00104j},

issn = {2632-8682},

year  = {2020},

date = {2020-08-01},

urldate = {2020-08-01},

journal = {RSC Med Chem},

volume = {11},

number = {8},

pages = {896--904},

abstract = {A series of compounds have been synthesized with a variety of substituents based on a three-carbon chain at the C9-position of 3-hydroxy--phenethyl-5-phenylmorphan (3-(2-phenethyl-2-azabicyclo[3.3.1]nonan-5-yl)phenol). Three of these were found to be μ-opioid receptor agonists in the inhibition of forskolin-induced cAMP accumulation assay and they did not recruit β-arrestin at all in the PathHunter assay and in the Tango assay. Compound  (3-((1,5,9)-2-phenethyl-9-propyl-2-azabicyclo[3.3.1]nonan-5-yl)phenol),  (3-((1,5,9)-9-(()-3-hydroxyprop-1-en-1-yl)-2-phenethyl-2-azabicyclo[3.3.1]nonan-5-yl)phenol), and  (3-((1,5,9)-9-(2-hydroxypropyl)-2-phenethyl-2-azabicyclo[3.3.1]nonan-5-yl)phenol) were partial μ-agonists. Two of them had moderate efficacies (   65%) and one had lower efficacy, and they were  5, 3, and 4 times more potent, respectively, than morphine . Computer simulations were carried out to provide a molecular basis for the high bias ratios of the C9-substituted 5-phenylmorphans toward G-protein activation.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid32517185,

title = {The Intriguing Effects of Substituents in the -Phenethyl Moiety of Norhydromorphone: A Bifunctional Opioid from a Set of },

author = {Meining Wang and Thomas C Irvin and Christine A Herdman and Ramsey D Hanna and Sergio A Hassan and Yong-Sok Lee and Sophia Kaska and Rachel Saylor Crowley and Thomas E Prisinzano and Sarah L Withey and Carol A Paronis and Jack Bergman and Saadet Inan and Ellen B Geller and Martin W Adler and Theresa A Kopajtic and Jonathan L Katz and Aaron M Chadderdon and John R Traynor and Arthur E Jacobson and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/32517185/},

doi = {10.3390/molecules25112640},

issn = {1420-3049},

year  = {2020},

date = {2020-06-01},

urldate = {2020-06-01},

journal = {Molecules},

volume = {25},

number = {11},

abstract = {(-)--Phenethyl analogs of optically pure -norhydromorphone were synthesized and pharmacologically evaluated in several in vitro assays (opioid receptor binding, stimulation of [S]GTPγS binding, forskolin-induced cAMP accumulation assay, and MOR-mediated β-arrestin recruitment assays). "Body" and "tail" interactions with opioid receptors (a subset of Portoghese's message-address theory) were used for molecular modeling and simulations, where the "address" can be considered the "body" of the hydromorphone molecule and the "message" delivered by the substituent (tail) on the aromatic ring of the -phenethyl moiety. One compound, N-p-chloro-phenethynorhydromorphone ((7aR,12bS)-3-(4-chlorophenethyl)-9-hydroxy-2,3,4,4a,5,6-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-7(7aH)-one, ), was found to have nanomolar binding affinity at MOR and DOR. It was a potent partial agonist at MOR and a full potent agonist at DOR with a δ/μ potency ratio of 1.2 in the ([S]GTPγS) assay. Bifunctional opioids that interact with MOR and DOR, the latter as agonists or antagonists, have been reported to have fewer side-effects than MOR agonists. The p-chlorophenethyl compound  was evaluated for its effect on respiration in both mice and squirrel monkeys. Compound  did not depress respiration (using normal air) in mice or squirrel monkeys. However, under conditions of hypercapnia (using air mixed with 5% CO), respiration was depressed in squirrel monkeys.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid32787282,

title = {Novel Vaccine That Blunts Fentanyl Effects and Sequesters Ultrapotent Fentanyl Analogues},

author = {Rodell C Barrientos and Eric W Bow and Connor Whalen and Oscar B Torres and Agnieszka Sulima and Zoltan Beck and Arthur E Jacobson and Kenner C Rice and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/32787282/},

doi = {10.1021/acs.molpharmaceut.0c00497},

issn = {1543-8392},

year  = {2020},

date = {2020-01-01},

urldate = {2020-01-01},

journal = {Mol Pharm},

volume = {17},

number = {9},

pages = {3447--3460},

abstract = {Active immunization is an emerging potential modality to combat fatal overdose amid the opioid epidemic. In this study, we described the design, synthesis, formulation, and animal testing of an efficacious vaccine against fentanyl. The vaccine formulation is composed of a novel fentanyl hapten conjugated to tetanus toxoid (TT) and adjuvanted with liposomes containing monophosphoryl lipid A adsorbed on aluminum hydroxide. The linker and hapten -phenyl--(1-(4-(3-(tritylthio)propanamido)phenethyl)piperidin-4-yl)propionamide were conjugated sequentially to TT using amine--hydroxysuccinimide-ester and thiol-maleimide reaction chemistries, respectively. Conjugation was facile, efficient, and reproducible with a protein recovery of >98% and a hapten density of 30-35 per carrier protein molecule. In mice, immunization induced high and robust antibody endpoint titers in the order of >10 against the hapten. The antisera bound fentanyl, carfentanil, cyclopropyl fentanyl, -fluorofentanyl, and furanyl fentanyl  with antibody-drug dissociation constants in the range of 0.36-4.66 nM. No cross-reactivity to naloxone, naltrexone, methadone, or buprenorphine was observed. , immunization shifted the antinociceptive dose-response curve of fentanyl to higher doses. Collectively, these preclinical results showcased the desired traits of a potential vaccine against fentanyl and demonstrated the feasibility of immunization to combat fentanyl-induced effects.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid32876947,

title = {Synthesis of a deuterated 6-AmHap internal standard for the determination of hapten density in a heroin vaccine drug product},

author = {Mariia Makarova and Rodell C Barrientos and Oscar B Torres and Gary R Matyas and Arthur E Jacobson and Agnieszka Sulima and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/32876947/},

doi = {10.1002/jlcr.3880},

issn = {1099-1344},

year  = {2020},

date = {2020-01-01},

urldate = {2020-01-01},

journal = {J Labelled Comp Radiopharm},

volume = {63},

number = {13},

pages = {564--571},

abstract = {A deuterated hapten was designed and synthesized that will be essential for a future study of residual hapten and stability of a hapten-protein conjugate. This hapten, 6-AmHap, was chosen for a heroin vaccine that is now slated for a Phase 1 clinical trial. A maleimide-thiol bioconjugation strategy was successfully applied to our heroin vaccine to connect the hapten 6-AmHap with an immunogenic carrier protein (tetanus toxoid, TT) through a trityl-protected 3-mercaptopropanamide linker. The antibodies induced by the vaccine have been found to have activity against several opioids, including heroin and its metabolites, and, importantly, leave alternate pain treatment medications such as methadone untouched. To the best of our knowledge, no other hapten for a heroin vaccine has been deuterated, yet this tool may prove to be of great importance in the study of residual hapten during product release and the long-term stability program of a hapten-protein conjugate as part of FDA regulatory requirements. Hydrocodone was the starting material for the synthesis of the deuterated 6-AmHap, with a stable amide at C6 and a 3-mercaptopropanamide linker attached at C3. The desired deuterated product was prepared as the disulfide, 3,3'-disulfanediylbis(N-((7S,7aR,12bS)-7-acetamido-3-[ H ]methyl)-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)propanamide), that could be easily reduced to form the needed hapten, N-((4aR,7S,7aR,12bS)-7-acetamido-3-[ H ]methyl]-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)-3-mercaptopropanamide.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid30913037b,

title = {Opioid-galanin receptor heteromers mediate the dopaminergic effects of opioids},

author = {Ning-Sheng Cai and César Quiroz and Jordi Bonaventura and Alessandro Bonifazi and Thomas O Cole and Julia Purks and Amy S Billing and Ebonie Massey and Michael Wagner and Eric D Wish and Xavier Guitart and William Rea and Sherry Lam and Estefanía Moreno and Verònica Casadó-Anguera and Aaron D Greenblatt and Arthur E Jacobson and Kenner C Rice and Vicent Casadó and Amy Hauck Newman and John W Winkelman and Michael Michaelides and Eric Weintraub and Nora D Volkow and Annabelle M Belcher and Sergi Ferré},

url = {https://pubmed.ncbi.nlm.nih.gov/30913037/},

doi = {10.1172/JCI126912},

issn = {1558-8238},

year  = {2019},

date = {2019-01-01},

urldate = {2019-01-01},

journal = {J Clin Invest},

volume = {129},

number = {7},

pages = {2730--2744},

abstract = {Identifying non-addictive opioid medications is a high priority in medical sciences, but μ-opioid receptors mediate both the analgesic and addictive effects of opioids. We found a significant pharmacodynamic difference between morphine and methadone that is determined entirely by heteromerization of μ-opioid receptors with galanin Gal1 receptors, rendering a profound decrease in the potency of methadone. This was explained by methadone's weaker proficiency to activate the dopaminergic system as compared to morphine and predicted a dissociation of therapeutic versus euphoric effects of methadone, which was corroborated by a significantly lower incidence of self-report of "high" in methadone-maintained patients. These results suggest that μ-opioid-Gal1 receptor heteromers mediate the dopaminergic effects of opioids that may lead to a lower addictive liability of opioids with selective low potency for the μ-opioid-Gal1 receptor heteromer, exemplified by methadone.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid30095301,

title = {Extracellular esterase activity as an indicator of endoplasmic reticulum calcium depletion},

author = {Kathleen A Trychta and Emily J Heathward and Agnieszka Sulima and Susanne Bäck and Mehdi Farokhnia and Christopher T Richie and Lorenzo Leggio and Kenner C Rice and Brandon K Harvey},

url = {https://pubmed.ncbi.nlm.nih.gov/30095301/},

doi = {10.1080/1354750X.2018.1490968},

issn = {1366-5804},

year  = {2018},

date = {2018-12-01},

urldate = {2018-12-01},

journal = {Biomarkers},

volume = {23},

number = {8},

pages = {756--765},

abstract = {CONTEXT: Endoplasmic reticulum (ER) calcium depletion is associated with diverse diseases, including cardiac, hepatic, and neurologic diseases.



OBJECTIVE: The aim of the present study was to identify and characterize an endogenous protein that could be used to monitor ER calcium depletion comparably to a previously described exogenous reporter protein.



MATERIALS AND METHODS: The use of a selective esterase-fluorescein diester pair allowed for carboxylesterase activity in extracellular fluid to be measured using a fluorescent readout. Cell culture media from three different cell lines, rat plasma, and human serum all possess quantifiable amounts of esterase activity.



RESULTS: Fluorescence produced by the interaction of carboxylesterases with a fluorescein diester substrate tracks with pharmacological and physiological inducers of ER calcium depletion. The fluorescence measured for in vitro and in vivo samples were consistent with ER calcium depletion being the trigger for increased esterase activity.



DISCUSSION: Decreased luminal ER calcium causes ER resident esterases to be released from the cell, and, when assessed concurrently with other disease biomarkers, these esterases may provide insight into the role of ER calcium homeostasis in human diseases.



CONCLUSION: Our results indicate that carboxylesterases are putative markers of ER calcium dysfunction.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid29675707,

title = {A rapid solution-based method for determining the affinity of heroin hapten-induced antibodies to heroin, its metabolites, and other opioids},

author = {Oscar B Torres and Alexander J Duval and Agnieszka Sulima and Joshua F G Antoline and Arthur E Jacobson and Kenner C Rice and Carl R Alving and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/29675707/},

doi = {10.1007/s00216-018-1060-4},

issn = {1618-2650},

year  = {2018},

date = {2018-06-01},

urldate = {2018-06-01},

journal = {Anal Bioanal Chem},

volume = {410},

number = {16},

pages = {3885--3903},

abstract = {We describe for the first time a method that utilizes microscale thermophoresis (MST) technology to determine polyclonal antibody affinities to small molecules. Using a novel type of heterologous MST, we have accurately measured a solution-based binding affinity of serum antibodies to heroin which was previously impossible with other currently available methods. Moreover, this mismatch approach (i.e., using a cross-reactive hapten tracer) has never been reported in the literature. When compared with equilibrium dialysis combined with ultra-performance liquid chromatography/tandem mass spectrometry (ED-UPLC/MS/MS), this novel MST method yields similar binding affinity values for polyclonal antibodies to the major heroin metabolites 6-AM and morphine. Additionally, we herein report the method of synthesis of this novel cross-reactive hapten, MorHap-acetamide-a useful analog for the study of heroin hapten-antibody interactions. Using heterologous MST, we were able to determine the affinities, down to nanomolar accuracies, of polyclonal antibodies to various abused opioids. While optimizing this method, we further discovered that heroin is protected from serum esterase degradation by the presence of these antibodies in a concentration-dependent manner. Lastly, using affinity data for a number of structurally different opioids, we were able to dissect the moieties that are crucial to antibody binding. The novel MST method that is presented herein can be extended to the analysis of any ligand that is prone to degradation and can be applied not only to the development of vaccines to substances of abuse but also to the analysis of small molecule/protein interactions in the presence of serum. Graphical abstract Strategy for the determination of hapten-induced antibody affinities using Microscale thermophoresis.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid29236495,

title = {A Stable Heroin Analogue That Can Serve as a Vaccine Hapten to Induce Antibodies That Block the Effects of Heroin and Its Metabolites in Rodents and That Cross-React Immunologically with Related Drugs of Abuse},

author = {Agnieszka Sulima and Rashmi Jalah and Joshua F G Antoline and Oscar B Torres and Gregory H Imler and Jeffrey R Deschamps and Zoltan Beck and Carl R Alving and Arthur E Jacobson and Kenner C Rice and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/29236495/},

doi = {10.1021/acs.jmedchem.7b01427},

issn = {1520-4804},

year  = {2018},

date = {2018-01-01},

urldate = {2018-01-01},

journal = {J Med Chem},

volume = {61},

number = {1},

pages = {329--343},

abstract = {An improved synthesis of a haptenic heroin surrogate 1 (6-AmHap) is reported. The intermediate needed for the preparation of 1 was described in the route in the synthesis of 2 (DiAmHap). A scalable procedure was developed to install the C-3 amido group. Using the Boc protectng group in 18 allowed preparation of 1 in an overall yield of 53% from 4 and eliminated the necessity of preparing the diamide 13. Hapten 1 was conjugated to tetanus toxoid and mixed with liposomes containing monophosphoryl lipid A as an adjuvant. The 1 vaccine induced high anti-1 IgG levels that reduced heroin-induced antinociception and locomotive behavioral changes following repeated subcutaneous and intravenous heroin challenges in mice and rats. Vaccinated mice had reduced heroin-induced hyperlocomotion following a 50 mg/kg heroin challenge. The 1 vaccine-induced antibodies bound to heroin and other abused opioids, including hydrocodone, oxycodone, hydromorphone, oxymorphone, and codeine.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid28314512,

title = {Modulation of opioid receptor affinity and efficacy via N-substitution of 9β-hydroxy-5-(3-hydroxyphenyl)morphan: Synthesis and computer simulation study},

author = {Phong M Truong and Sergio A Hassan and Yong-Sok Lee and Theresa A Kopajtic and Jonathan L Katz and Aaron M Chadderdon and John R Traynor and Jeffrey R Deschamps and Arthur E Jacobson and Kenner C Rice},

url = {https://pubmed.ncbi.nlm.nih.gov/28314512/},

doi = {10.1016/j.bmc.2017.02.064},

issn = {1464-3391},

year  = {2017},

date = {2017-01-01},

urldate = {2017-01-01},

journal = {Bioorg Med Chem},

volume = {25},

number = {8},

pages = {2406--2422},

abstract = {The enantiomers of a variety of N-alkyl-, N-aralkyl-, and N-cyclopropylalkyl-9β-hydroxy-5-(3-hydroxyphenyl)morphans were synthesized employing cyanogen bromide and KCO to improve the original N-demethylation procedure. Their binding affinity to the μ-, δ-, and κ-opioid receptors (ORs) was determined and functional (GTPS) assays were carried out on those with reasonable affinity. The 1R,5R,9S-enantiomers (1R,5R,9S)-(-)-5-(3-hydroxyphenyl)-2-(4-nitrophenethyl)-2-azabicyclo[3.3.1]nonan-9-ol (1R,5R,9S-16), (1R,5R,9S)-(-) 2-cinnamyl-5-(3-hydroxyphenyl)-2-azabicyclo[3.3.1]nonan-9-ol (1R,5R,9S-20), and (1R,5R,9S)-(-)-5-(3-hydroxyphenyl)-2-(4-(trifluoromethyl)phenethyl)-2-azabicyclo[3.3.1]nonan-9-ol (1R,5R,9S-15), had high affinity for the μ-opioid receptor (e.g., 1R,5R,9S-16: Ki=0.073, 0.74, and 1.99nM, respectively). The 1R,5R,9S-16 and 1R,5R,9S-15 were full, high efficacy μ-agonists (EC=0.74 and 18.5nM, respectively) and the former was found to be a partial agonist at δ-OR and an antagonist at κ-OR, while the latter was a partial agonist at δ-OR and κ-OR in the GTPS assay. The enantiomer of 1R,5R,9S-16, (+)-1S,5S,9R-16 was unusual, it had good affinity for the μ-OR (Ki=26.5nM) and was an efficacious μ-antagonist (Ke=29.1nM). Molecular dynamics simulations of the μ-OR were carried out with the 1R,5R,9S-16 μ-agonist and the previously synthesized (1R,5R,9S)-(-)-5-(9-hydroxy-5-(3-hydroxyphenyl-2-phenylethyl)-2-azabicyclo[3.3.1]nonane (1R,5R,9S-(-)-NIH 11289) to provide a structural basis for the observed high affinities and efficacies. The critical roles of both the 9β-OH and the p-nitro group are elucidated, with the latter forming direct, persistent hydrogen bonds with residues deep in the binding cavity, and the former interacting with specific residues via highly structured water bridges.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid29263870,

title = {Heroin-HIV-1 (H2) vaccine: induction of dual immunologic effects with a heroin hapten-conjugate and an HIV-1 envelope V2 peptide with liposomal lipid A as an adjuvant},

author = {Oscar B Torres and Gary R Matyas and Mangala Rao and Kristina K Peachman and Rashmi Jalah and Zoltan Beck and Nelson L Michael and Kenner C Rice and Arthur E Jacobson and Carl R Alving},

url = {https://pubmed.ncbi.nlm.nih.gov/29263870/},

doi = {10.1038/s41541-017-0013-9},

issn = {2059-0105},

year  = {2017},

date = {2017-01-01},

urldate = {2017-01-01},

journal = {NPJ Vaccines},

volume = {2},

pages = {13},

abstract = {A synthetic heroin analog (MorHap) and a synthetic 42 amino acid V2 loop peptide from A/E strain of HIV-1 gp120 envelope protein that was previously used in a successful phase III vaccine trial were constructed as antigens together with liposomes containing monophosphoryl lipid A as an adjuvant, to explore the feasibility of producing a dual use vaccine both for treatment of heroin addiction and prevention of HIV-1 infection among injection drug users. The V2 peptide was tethered by a palmitoyl fatty acyl tail embedded in the liposomal lipid bilayer, and the heroin analog was conjugated to tetanus toxoid as a carrier protein that was mixed with the adjuvant. Upon comparison of a linear V2 peptide with a cyclic peptide, differences were found in the secondary configurations by circular dichroism, with the tethered cyclic peptide (palm-cyclic peptide) entirely in a random coil, and the tethered linear V2 peptide (palm-linear V2 peptide) entirely in a beta-sheet. Upon immunization of mice, palm-cyclic peptide induced anti-cyclic peptide endpoint titers >10 and was considered to be a better immunogen overall than palm-linear V2 peptide for inducing antibodies to gp120 and gp70-V1V2. The antibodies also inhibited the binding of V2 peptide to the HIV-1 αβ integrin receptor. Antibody titers to MorHap, even with the presence of injected cyclic peptide, were very high, and resulted in inhibition of the hyper-locomotion and antinociception effects of injected heroin. From these initial experiments, we conclude that with a potent adjuvant and mostly synthetic constituents, a vaccine directed to heroin and HIV-1 (H2 vaccine) could be a feasible objective.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid26677020,

title = {A simple nonradioactive method for the determination of the binding affinities of antibodies induced by hapten bioconjugates for drugs of abuse},

author = {Oscar B Torres and Joshua F G Antoline and Fuying Li and Rashmi Jalah and Arthur E Jacobson and Kenner C Rice and Carl R Alving and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/26677020/},

doi = {10.1007/s00216-015-9223-z},

issn = {1618-2650},

year  = {2016},

date = {2016-02-01},

urldate = {2016-02-01},

journal = {Anal Bioanal Chem},

volume = {408},

number = {4},

pages = {1191--1204},

abstract = {The accurate analytical measurement of binding affinities of polyclonal antibody in sera to heroin, 6-acetylmorphine (6-AM), and morphine has been a challenging task. A simple nonradioactive method that uses deuterium-labeled drug tracers and equilibrium dialysis (ED) combined with ultra performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) to measure the apparent dissociation constant (K d) of antibodies to 6-AM and morphine is described. The method can readily detect antibodies with K d in the low nanomolar range. Since heroin is rapidly degraded in sera, esterase inhibitors were included in the assay, greatly reducing heroin hydrolysis. MS/MS detection directly measured the heroin in the assay after overnight ED, thereby allowing the quantitation of % bound heroin in lieu of K d as an alternative measurement to assess heroin binding to polyclonal antibody sera. This is the first report that utilizes a solution-based assay to quantify heroin-antibody binding without being confounded by the presence of 6-AM and morphine and to measure K d of polyclonal antibody to 6-AM. Hapten surrogates 6-AcMorHap, 6-PrOxyHap, MorHap, DiAmHap, and DiPrOxyHap coupled to tetanus toxoid (TT) were used to generate high affinity antibodies to heroin, 6-AM, and morphine. In comparison to competition ED-UPLC/MS/MS which gave K d values in the nanomolar range, the commonly used competition enzyme-linked immunosorbent assay (ELISA) measured the 50% inhibition concentration (IC50) values in the micromolar range. Despite the differences in K d and IC50 values, similar trends in affinities of hapten antibodies to heroin, 6-AM, and morphine were observed by both methods. Competition ED-UPLC/MS/MS revealed that among the five TT-hapten bioconjugates, TT-6-AcMorHap and TT-6-PrOxyHap induced antibodies that bound heroin, 6-AM, and morphine. In contrast, TT-MorHap induced antibodies that poorly bound heroin, while TT-DiAmHap and TT-DiPrOxyHap induced antibodies either did not bind or poorly bound to heroin, 6-AM, and morphine. This simple and nonradioactive method can be extended to other platforms, such as oxycodone, cocaine, nicotine, and methamphetamine for the selection of the lead hapten design during substance abuse vaccine development.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Selfridge2015,

title = {Structure-Activity Relationships of (+)-Naltrexone-Inspired Toll-like Receptor 4 (TLR4) Antagonists.},

author = {Brandon R Selfridge and Xiaohui Wang and Yingning Zhang and Hang Yin and Peter M Grace and Linda R Watkins and Arthur E Jacobson and Kenner C Rice},

url = {https://www.ncbi.nlm.nih.gov/pubmed/26010811},

doi = {10.1021/acs.jmedchem.5b00426},

issn = {1520-4804 (Electronic); 0022-2623 (Linking)},

year  = {2015},

date = {2015-06-25},

journal = {J Med Chem},

volume = {58},

number = {12},

pages = {5038--5052},

abstract = {Activation of Toll-like receptors has been linked to neuropathic pain and opioid dependence. (+)-Naltrexone acts as a Toll-like receptor 4 (TLR4) antagonist and has been shown to reverse neuropathic pain in rat studies. We designed and synthesized compounds based on (+)-naltrexone and (+)-noroxymorphone and evaluated their TLR4 antagonist activities by their effects on inhibiting lipopolysaccharide (LPS) induced TLR4 downstream nitric oxide (NO) production in microglia BV-2 cells. Alteration of the N-substituent in (+)-noroxymorphone gave us a potent TLR4 antagonist. The most promising analog, (+)-N-phenethylnoroxymorphone ((4S,4aR,7aS,12bR)-4a,9-dihydroxy-3-phenethyl-2,3,4,4a,5,6-hexahydro-1H-4,12-meth anobenzofuro[3,2-e]isoquinolin-7(7aH)-one, 1j) showed approximately 75 times better TLR-4 antagonist activity than (+)-naltrexone, and the ratio of its cell viability IC50, a measure of its toxicity, to TLR-4 antagonist activity (140 muM/1.4 muM) was among the best of the new analogs. This compound (1j) was active in vivo; it significantly increased and prolonged morphine analgesia.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{pmid25970207,

title = {Efficacy, but not antibody titer or affinity, of a heroin hapten conjugate vaccine correlates with increasing hapten densities on tetanus toxoid, but not on CRM_{197} carriers},

author = {Rashmi Jalah and Oscar B Torres and Alexander V Mayorov and Fuying Li and Joshua F G Antoline and Arthur E Jacobson and Kenner C Rice and Jeffrey R Deschamps and Zoltan Beck and Carl R Alving and Gary R Matyas},

url = {https://pubmed.ncbi.nlm.nih.gov/25970207/},

doi = {10.1021/acs.bioconjchem.5b00085},

issn = {1520-4812},

year  = {2015},

date = {2015-06-01},

urldate = {2015-06-01},

journal = {Bioconjug Chem},

volume = {26},

number = {6},

pages = {1041--1053},

abstract = {Vaccines against drugs of abuse have induced antibodies in animals that blocked the biological effects of the drug by sequestering the drug in the blood and preventing it from crossing the blood-brain barrier. Drugs of abuse are too small to induce antibodies and, therefore, require conjugation of drug hapten analogs to a carrier protein. The efficacy of these conjugate vaccines depends on several factors including hapten design, coupling strategy, hapten density, carrier protein selection, and vaccine adjuvant. Previously, we have shown that 1 (MorHap), a heroin/morphine hapten, conjugated to tetanus toxoid (TT) and mixed with liposomes containing monophosphoryl lipid A [L(MPLA)] as adjuvant, partially blocked the antinociceptive effects of heroin in mice. Herein, we extended those findings, demonstrating greatly improved vaccine induced antinociceptive effects up to 3% mean maximal potential effect (%MPE). This was obtained by evaluating the effects of vaccine efficacy of hapten 1 vaccine conjugates with varying hapten densities using two different commonly used carrier proteins, TT and cross-reactive material 197 (CRM197). Immunization of mice with these conjugates mixed with L(MPLA) induced very high anti-1 IgG peak levels of 400-1500 μg/mL that bound to both heroin and its metabolites, 6-acetylmorphine and morphine. Except for the lowest hapten density for each carrier, the antibody titers and affinity were independent of hapten density. The TT carrier based vaccines induced long-lived inhibition of heroin-induced antinociception that correlated with increasing hapten density. The best formulation contained TT with the highest hapten density of ≥30 haptens/TT molecule and induced %MPE of approximately 3% after heroin challenge. In contrast, the best formulation using CRM197 was with intermediate 1 densities (10-15 haptens/CRM197 molecule), but the %MPE was approximately 13%. In addition, the chemical synthesis of 1, the optimization of the conjugation method, and the methods for the accurate quantification of hapten density are described. },

keywords = {},

pubstate = {published},

tppubtype = {article}

}

@article{Suzuki2015,

title = {Chiral resolution and absolute configuration of the enantiomers of the psychoactive },

author = {Masaki Suzuki and Jeffrey R Deschamps and Arthur E Jacobson and Kenner C Rice},

url = {https://www.ncbi.nlm.nih.gov/pubmed/25727807},

doi = {10.1002/chir.22423},

issn = {1520-636X (Electronic); 0899-0042 (Linking)},

year  = {2015},

date = {2015-04-01},

journal = {Chirality},

volume = {27},

number = {4},

pages = {287--293},

address = {Drug Design and Synthesis Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland.},

abstract = {Illicit rac-MDPV (3,4-methylenedioxypyrovalerone), manufactured in clandestine labs, has become widely abused for its cocaine-like stimulant properties. It has recently been found as one of the toxic materials in the so-called "bath salts," producing, among other effects, psychosis and tachycardia in humans when introduced by any of the several routes of administration (e.g., intravenous, oral, etc.). The considerable toxicity of this "designer drug" probably resides in one of the enantiomers of the racemate. In order to obtain a sufficient amount of the enantiomers of rac-MDPV to determine their activity, we improved the known synthesis of rac-MDPV and found chemical resolving agents, (+)- and (-)-2'-bromotetranilic acid, that gave the MDPV enantiomers in >96% enantiomeric excess as determined by (1) H nuclear magnetic resonance and chiral high-performance liquid chromatography. The absolute stereochemistry of these enantiomers was determined by single-crystal X-ray diffraction studies.},

keywords = {},

pubstate = {published},

tppubtype = {article}

}