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- 1. The Synthesis ofNPFF Receptor Ligand A dual-acting compound with opioid agonist & neuropeptide FF antagonist
- 2. Introduction Caroline Morris Hernando, MS Universityof Mississippi Undergraduate Junior B.S in Pharmaceutical Sciences, pursuing Pharm.D
- 3. Table of Contents 1.Background 2. Projected Solution 3. Methodology 4. Conclusion 5. Personal Development 6. Acknowledgements 7. Questions
- 4. Background • Chronic painaffects 116 million Americans per year 1 • Opioid drugs are the primary prescribed pain- killers 1O’Reilly, Kevin B. American Medical News. 2011
- 5. Background • Opioid drugshave several side effects: - digestive problems2 - addiction3 - tolerance3 As the dosage increases, the side effects increase until the point that the side effects outweigh the benefits 2Chan, Lingtak-Neander. Nutrition Issues in Gastroenterology. 2008 3Stuckert, Jeffery. Psych Central. 2011
- 6. Projected Solution • Previousresearch has discovered that the Neuropeptide FF antagonist receptors can prevent morphine-induced tolerance • The use of a dual-acting compound with Opioid & NPFF receptors looks promising: -opioid drug use without tolerance
- 7. Projected Solution 1. VBJ192works - warm-water immersion test
- 8. Projected Solution 2. Blood-BrainBarrier 3. Structure-Activity Relationship (SAR)
- 9. Projected Solution • Amidinefunctional group can penetrate the blood- brain barrier • Similarities to VBJ192
- 10. Methodology 1. Synthesis ofBenzyltriphenylphosphonium salt A: 1 mole, B: 1 mole, C: 97% yield A B C
- 11. Methodology 2. Wittig Reaction AB A: 34.62mmol, LDA: 38.06mmol, 1-benzylpiperidin-4-one: 6.7mL, B: 42.2% yield
- 12. Methodology 3. Bromination A B A:5.58mmol, Br2: 8.37mmol, NaOH: 21.63mmol, B: 80.7% yield
- 13. Methodology 4. Suzuki Coupling AB C A: 5.23mmol, B: 6.28mmol, Pd(PPh3)4: 0.1205g, K2CO3: 13.09mmol, C: 52.1% yield
- 14. Methodology 5. Garigipati Reaction AB A: 0.275mmol, NH4Cl: 0.495mmol, Al(CH3)3: 0.458mmol, B: 56% yield
- 15. Methodology 1. Filtration 2.Column Chromatography
- 16. Methodology • Because thissynthesis required multiple steps, after each step, verification had to take place in before starting the next reaction -in order to determine it was actually the desired product -make sure there were no impurities
- 17. Methodology Thin Layer Chromatography(TLC) - used to determine Rf value Rf value = distance product traveled distance solvent traveled = 6.25 cm 8.00 cm = 0.781
- 18. Methodology TLC is alsoused to make comparisons: -the compound is the same across, verifying that the different tubes from column chromatography contain the same compound.
- 19. Methodology -Bromination was successful!-Suzuki was successful! Mass Spectrometry (MS) -verifies the mass of the compound
- 20. Conclusion • We successfullymodified VBJ192 into an amidine derivative • NMR verified • Pharmacological evaluation will be performed on this compound
- 21. Personal Development 1. Honedresearch/lab skills 2. Delved into Medicinal Chemistry 3. Application of prior information 4. Patience
- 22. Acknowledgements Dr. Christopher McCurdy Dr.Coco Kapanda Kelsey Luecke Dr. Velvet Journigan Dr. Stephen Cutler COBRE Grant P20GM104932
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Editor's Notes
- #5 http://www.amednews.com/article/20111017/profession/310179937/2/ http://www.iom.edu/activities/publichealth/painresearch/pain-care-improvements-recommended-in-iom-report.aspx ***So what’s the big deal? We have Americans in pain, and we have opioids to fight their pain…
- #6 2. http://www.medicine.virginia.edu/clinical/departments/medicine/divisions/digestive-health/nutrition-support-team/nutrition-articles/ChanArticle.pdf --opioid receptors are also found in the gastrointestinal tract; causes issues with the normal motility and secretory functions within the tract 3. http://psychcentral.com/lib/opioid-dependence-and-withdrawal/0008507 --Opioid addiction is a disease of the brain. Repeated use of an opioid leading to opioid dependence causes long-term changes in both the structure (the architecture of the brain) and the way the brain functions (the biochemistry of the brain). Many use it outside of the prescription directions. 4.SAME AS 3 Tolerance: need more of the opioid to get the desired effect the GI issues become worse with the increase in dosage – making the side effects worse than the pain-relieving benefits Opioid drugs, such as morphine, have to be increased overtime to continue delivering an adequate amount of pain relief
- #7 -- previous reseach shows that we do have a possible solution. If we act on the NPFF system in coorelation with the opioid system, then tolerance will not develop. No tolerance means no increased dosages. No increased dosages means less addiction and less harm to the GI tract.
- #8 --Previous research within our own lab shows that compound VBJ192 works due to testing performed on mice. However, it only works when injected directly into the brain. Noone wants to inject shots into their brain. Need to find a drug that will work orally. That’s where the BBB comes into play. The BBB is very picky. Does not work if something is 1.
- #9 That’s where the BBB comes into play. The BBB is very picky. Does not work if something is too highly charged, too large, not lipid soluble.. Also, if it is too basic. AND, we know that 5 Nitrogens within a structure is the max number of nitrogens that can enter the BBB. Therefore, we would like to use SAR. Once we change different parts, we can try & enter the BBB. SAR will allow us to determine what part is necessary to have the desired function As well as help us change structures to one that will pass BBB more efficiently.
- #10 For my compound, we used SAR and changed from guanidine to aminidine. WHY? amidine functional group has been shown to be able to penetrate the Blood-Brain Barrier (BBB) from peripheral administration when it has been substituted for the guanidine functional group. Hoping the rest of the compound will work as VBJ192 did, but be able to penetrate BBB so we can make oral drugs.
- #11 Bromobenzene was added to triphenylphosphine, was dissolved in toluene, and refluxed at toluene’s BP overnight to yield the benzyltriphenylphosphonuim salt, which Is needed to proceed with the Wittig Reaction. 97% yield
- #12 Used the salt made, combined it with benzylpiperidone & refluxed overnight in a solution of LDA. LDA was used in order to produce the YLIDE. (+ and - charge next to each other) It was very important, at the step, to perform the reflux under argon, due to LDA’s reactivity with our air. LDA will take a Hydrogen, which destroys LDA’s ability, which stops the reaction from taking place. Produced our desired compound, 1-benzyl-4-benzylidenepiperidine, and by product triphosphoniumoxide. After separating the two we proceeded.. 42.2% yield
- #13 To bromination. This was a cool step. The bromine was a dark orange color with fuming orange vapors. --let it stir, heated & added NaOH and MeOH. Did purification to just have desired product without any starting materials. 80.7% yield
- #14 We took our brominated product and reacted it with cynaopheynlboronic acid and tetrakis(triphenylphosphine) palladium. The palladium is air sensitive (used argon) as well as light sensitive. (Also, side note – makes a really pretty neon color show when put under UV light for TLC). So, we had to wrap the flask in foil to prevent the reactants from being exposed to light as it refluxed over night. This reaction added the phenyl group with the cyano to our compound. 52.1% yield
- #15 Then, came the last part of the synthesis. Converting the cyano group into amindine. THEORETICAL YIELD:
- #16 Gravity filtration in order to separate the different compounds of the mixture. Used this to separate the desired product from byproduct in Wittig reaction. Chromotography to separate the compound for purity: use different mobile phases based of polarity that allowed for separation. We didn’t use pressure. Very time-consuming.
- #17 So, we made the compound, purified it, then verified it was the actual desired product in a purified form.
- #19 Different vials from Chromography. Used a sample of what we new was the compound (from previous column)
- #20 -verifies the mass of the compound -shows that there aren’t detectable impurities because of single peak
- #21 . If the in vitro assays are in close agreement with VBJ-192, MCM1 will be investigated in vivo for antinociceptive activity through peripheral administration in Core D of the COBRE program, followed by investigation at Torrey Pines for acute antinociceptive tolerance.
- #22 --learned to be patient because chemistry doesn’t work on a planned schedule --you will mess up; it’s a part of the practice. Ex. Dropped stir bar into chemical waste bucket.