Tonix Pharmaceuticals Holding Corp. · NASDAQ:TNXP
Tonix Pharmaceuticals — Technical Presentation (January 2023)
40 slides from Tonix Pharmaceuticals's technical presentation, Jan 31, 2023
Slide 1: Primary vs Secondary Sex Hormones and Migraine
Primary vs Secondary Sex Hormones and Migraine 1
Slide 2: Disclosures
Disclosures 2 Professor David C Yeomans, Director of Pain Research, Stanford Medical School • SiteOne Therapeutics – Founder • NewBio – Founder • Tonix Pharmaceuticals - Consultant • Nalu Medical – SAB chair • Cytonics – SAB chair • Circuit Therapeutics – Consultant • Endo Pharmaceuticals - Consulta
Slide 3: Sex Hormones
Sex Hormones 3 Sex hormones are critical regulators of sex - related characteristics and behaviors Primary: • Progesterone • Estrogens • Androgens Secondary: • Prolactin • Oxytocin
Slide 4: Estrogens and Migraine
The “Estrogen withdrawal hypothesis”, developed by Somerville and colleagues in 1972, postulates that attacks of menstrual migraine are triggered by the decrease in estrogen levels preceding menstruation. Hypothesized pathology: • A drop in estrogen may cause an increased sensitivity to prostaglandi
Slide 5: Estrogens are generally ineffective in migraine prevention RCTs
However, estrogens are generally ineffective in in migraine prevention RCTs 5 RCT (35 pts): Percutaneous Estradiol reduced migraine frequency by 22% compared to placebo but increased migraines as soon as drug stopped (MacGregor et al., 2006) RCT (27 pts): Percutaneous Estradiol no significant effect
Slide 6: Progesterone is analgesic in the trigeminal system
Progesterone is analgesic in the trigeminal system Progesterone Receptors are co - Expressed with Nav1.7 in trigeminal ganglia neurons Progesterone decreases sodium currents in mouse trigeminal ganglia neurons Bi et al., 2020 6
Slide 7: However, Progesterone is minimally effective
However, Progesterone is minimally effective NO RCTs Warhurst et al., 2017 7
Slide 8: Testosterone
Testosterone 8 No RCT but pilot study showing strong effects of testosterone implant on 27 female migraineurs with symptoms of androgen insufficiency . Compared to pre - treatment, testosterone produced a significant decrease in migraine severity with 74 % reporting severity scores of zero (Glaser e
Slide 9: Prolactin receptors on female, but not male mouse dural afferents
Prolactin receptors on female, but not male mouse dural afferents 9 Avona et al., 2021
Slide 10: Oxytocin is a polypeptide hormone/neurotransmitter
Oxytocin is a 9 amino acid polypeptide hormone/neurotransmitter which is made in the hypothalamus and secreted both into the systemic circulation and into certain CNS sites 10
Slide 11: Pregnancy increases oxytocin and decreases migraine frequency
Pregnancy increases oxytocin and decreases migraine frequency 90 80 70 60 50 40 30 20 10 0 Migraine Remission Rate (% ) Adapted from Hoshiyama, 2012 Kuwabara et al., 1987 11
Slide 12: Breast feeding releases oxytocin and prevents migraines
Breast feeding releases oxytocin and prevents migraines 9 8 7 6 5 4 3 2 Baseline Bottle Fed Feeding Breast Fed Plasma Oxytocin (pg/mL) 0 20 40 60 80 100 1 2 6 Weeks Post - Partum Migraine Recurrence Rate (%) Grewen et al, 2010 12 Bottle Fed Breast Fed Hoshiyama, 2012
Slide 13: Oxytocin receptor expression on trigeminal ganglia neurons
Oxytocin receptor expression on trigeminal ganglia neurons 13
Slide 14: Oxytocin receptors co-express CGRP in human trigeminal ganglia neurons
CGRP 14 OTR CGRP + OTR only Oxytocin receptors co - express CGRP in human trigeminal ganglia neurons
Slide 15: Oxytocin Inhibits Firing of Rat Trigeminal CGRP Neurons
Oxytocin Inhibits Firing of Rat Trigeminal (TG) CGRP Neurons Oxytocin Receptors Co - Localize with CGRP in Trigeminal Ganglia Neurons Oxytocin Receptors = red CGRP = green Oxytocin Inhibits Electrically Evoked Activity of Trigeminal Ganglia Neurons 10 μM Oxytocin Control Wash 15
Slide 16: Factors varying over the menstrual cycle control oxytocin activity
Multiple factors that vary over the menstrual cycle have been shown to control oxytocin activity 16
Slide 17: 1. IL-6 is low during menstrual migraine
1. IL - 6 is low during menstrual migraine 17
Slide 18: The OTR gene promoter has response elements for IL-6
18 The OTR gene promoter has multiple response elements for the inflammatory cytokine IL - 6 Nucletide Sequence (with cleavage Sites) 1 - 250 - 750 - 1000 - 403 - 500 - 722 - 909 - 1203 - 1250 hOTR gene Promoter Consensus Response Elements Transcription start site NF - IL6 - 181 NF - IL6 - 777 NF -
Slide 19: Oxytocin Preferentially Transported Through Trigeminal System After Nasal Delivery
Trigeminal nucleus oxytocin immunofluorescence I II III IV V Autoradiograms of trigeminal ganglia Oxytocin Is Preferentially Transported Throughout the Trigeminal System After Nasal Delivery (not systemically distributed) Oxytocin tissue levels (nM) after intranasal administration* TRIGEMINAL NERVE
Slide 20: Inflammation increases trigeminal Oxytocin receptor expression
Inflammation increases trigeminal Oxytocin receptor expression *# *# *# *# *# * * * * * Baseline 20
Slide 21: Ibuprofen blocks analgesic efficacy of nasal oxytocin after incision
0 35 30 25 20 15 10 5 1 2 3 Pain threshold (g) Saline + Ibuprofen After incision, the analgesic efficacy of nasal oxytocin is blocked by ibuprofen – which blocks IL - 6 production Pre - incision Post - incision Oxytocin Oxytocin + Ibuprofen 21 Post - treatment
Slide 22: IL-6 induces upregulation of oxytocin receptors in human ganglia
15 10 5 0 20 OTR Area of OTR+ve staining ( m 2 ) ٓٓٓٓ In vitro treatment of human ganglia with IL - 6 induces upregulation of oxytocin receptors Control 22 IL - 6 treated
Slide 23: Nasal delivery of oxytocin to the trigeminal system of humans
Nasal delivery of oxytocin to the trigeminal system of humans 23
Slide 24: Oxytocin in human TG after nasal application
13 N - Oxytocin in human TG after nasal application 24
Slide 25: Pilot Clinical Study: Nasal Oxytocin Reduces Pain In Chronic Migraineurs
Pilot Clinical Study: Nasal Oxytocin Reduces Pain In Chronic Migraineurs * = p< 0.05 compared to placebo Z - test of 2 proportions 20 10 0 30 40 50 60 0.5 Hrs 1.0 Hrs 2.0 Hrs Time after dosing 4.0 Hrs 24 Hrs Placebo OT Overall OT - NSAID OT + NSAID * 25 p = 0.0195 p = 0.0326 * * p = 0.0503 p = 0.019
Slide 26: 2. Estrogen is low during menstrual migraine
2. Estrogen is low during menstrual migraine 26
Slide 27: Plasma OT level is modulated by exogenous estrogen
27 Plasma OT level is modulated by exogenous estrogen Chiodera et al., 1991 Plasma oxytocin levels following oral estrogen (1 mg) Hours following estrogen administration
Slide 28: Increase in brain OXTR induced by estrogen
Increase in brain OXTR induced by estrogen 28
Slide 29: OT more potent inhibiting TG excitability in females
OT is more potent in inhibiting TG excitability in females, but with estrogen pretreatment, male potency is similar to female - 10 40 30 20 10 0 0 200 800 1000 400 600 OT dose (nM) Hyperpolarizatin (mv) male estogen ( - ) male estrogen (+) female 29
Slide 30: 3. Magnesium ion plasma level is low during menstrual migraine
3. Magnesium ion plasma level is low during menstrual migraine 30
Slide 31: Serum Mg++ is lower in migraineurs
Attack Assarzadegan et al., 2016 31 0 1 0.5 2.5 2 1.5 Control Interictal Serum Mg++ (mg/DL) * Serum Mg++ is lower in migraineurs
Slide 32: Reduced basal and attack-dependent serum Mg++ in migraineurs
Reduced basal and attack - dependent serum Mg++ in migraineurs Sarchielli et al., 1991 0.6 0.5 0.4 32 0.7 0.8 1 0.9 Interictal Attack • Odds of migraine is increased 35.3 times when serum levels of magnesium reached below the normal level.
Slide 33: Binding site for Mg2+ between OT and OTR
Binding site for Mg 2+ between OT and OTR 33
Slide 34: Mg2+ enhanced OT-driven desensitization of rat TG
Addition of Mg 2+ enhanced OT - driven desensitization of rat TG 34
Slide 35: Mg2+ enhanced OT-driven craniofacial analgesia
Addition of Mg 2+ enhanced OT - driven craniofacial analgesia 35
Slide 36: Mg2+ enhanced OT-driven increase of rheobase of human DRG
0XY + 0.5mM Mg2+ 0XY + 1.75mM Mg2+ Washout Baseline 20mV 100ms ∆ Rheobase Addition of Mg 2+ enhanced OT - driven increase of rheobase of human 36 DRG
Slide 37: Mg2+ enhanced OT-driven decrease in AP firing in human DRG
Baseline 37 0XY + 0.5mM Mg2+ 0XY + 1.75mM Mg2+ Addition of Mg 2+ enhanced OT - driven decrease in AP following in human DRG
Slide 38: Summary: Sex hormone modulation of migraine
Summary: Sex hormone modulation of migraine 38 • Evidence for Direct modulation of TG excitability by primary sex hormones in not strong • Evidence for extended sex hormones (Prolactin, oxytocin) robust • Oxytocin effects are modulated by several endogenous factors that vary over the menstrual cycle
Slide 39: Alternatively, hugging, massage, sex, looking at your dog could help prevent migraines
Alternatively, hugging, massage, sex, looking at your dog could help prevent migraines 39
Slide 40: Thank you!
Thank you! 40
Tonix Pharmaceuticals Holding — Technical Presentation (January 2023) on 6ix