Mediators of Inflammation
`
`223-228 2000
`
`Effect of ricinoleic acid in acute and
`subchronic experimental models of
`inflammation
`
`Celme Vieiral Stefano EvangeIista2
`Rocco CiriIIo1 Annalisa Lippi1 Carlo Alberto Maggi1
`and Stefano Manzini2
`
`and 2Preclinical
`Departments of 1Pharmacology
`Development Menarini Ricerche spa Pomezia
`Roma and Firenze Italy
`
`CACorrespondthg Author
`Tel 39 055 5680519
`Fax 39 055 5680510
`E-mail sevangelista@menarini-ricerche .i
`
`CV was
`de Riberào Preto
`
`visiting scientist
`U.S.P Brazil
`
`from Faculdade
`
`de Medicine
`
`Research Paper
`
`OBSERVATIONAL studies indicate that topical application
`of ricinoleic add RA the main component of castor
`oil exerts remarkable analgesic and anti-inflamma
`has
`tory effects Pharmacological
`characterization
`shown similarities between the effects of RA and those
`interaction of this
`of capsaidn suggesting
`potential
`drug on sensory neuropeptide-mediated
`neurogenic
`inflammation The aim of this study was to assess RA
`anti-inflammatory activities in comparison with cap
`saicin in several models of acute and subchronic
`inflammation The acute inflammation was inducedby
`intradermal injection of carrageenan in the mouse or
`by histamine in the guinea-pig eyelid In either
`experiment the extent of the oedema thickness was
`measured Subchronic oedema was induced by com
`plete Freunds adjuvant
`injection in the ventral right
`paw of mice Tissue substance SP was measured in
`the carrageenan experiments by radioimmunoassay
`RIA It was found that the acute topical application of
`RA 0.9 mg/mouse or capsaidn 0.09 mg/mouse sig
`nificantly increased the mouse paw oedema induced
`by carrageenan while an 8-thy repeated topical
`treatment with the same doses of both compounds
`marked inhibition of carrageenan
`resulted in
`induced paw oedema matched by
`reduction in SP
`tissue levels Similar effects were found against hista
`mine-induced eyelid oedema in guinea-pigs after acute
`or repeated application of RA or capsaicin RA and
`capsaidn given for 13 weeks reduced the established
`oedema induced by Freunds adjuvant
`subchronic
`model of inflammation particularly if given by the
`intradermal route Either in mouse paw or in guinea-
`pig eyelid capsaicin but not RA by itself produced
`slight hyperemia and activation of
`behavioural
`response e.g scratching of the eyelids On the basis
`new
`results RA may be seen as
`of the present
`non-pungent anti-inflammatory agent
`capsaidn-like
`suitable for peripheral application
`
`Key words Cap saicin Carrageenan Castor oil
`SP
`tion Ricinoleic acid Substance
`
`Inflamma
`
`Introduction
`
`Ricinoleic acid RA
`12-hydroxy9-octadece-
`the main component of castor oil
`noic acid is
`accounting for about 90% of
`the total Preliminary
`laboratories indicated that RA
`experiments in our
`known for its laxative properties could have
`pro-
`or anti-inflammatory action following acute or repeti
`tive local application respectively These activities
`closely resemble those previously described for cap
`vanilloids
`saicin or capsaicinoid
`called
`analogues
`On the other
`olvanil
`resiniferatoxin scutigeral.2
`hand part of the RA chemical structure has been used
`basis for the development of novel capsaicin-like
`as
`The cloning of the vanilloid receptor
`compounds.5
`termed YR1 is very recent6 and it has
`
`ion channel
`
`ISSN 0962-9351 print/ISSN 1466-1861 online/00/050223-06
`001 10.1080/09629350020025737
`
`been shown that YR1 is activated by capsaicin
`and
`number of other capsaicin-like drugs vanilloid recep
`through the
`tor agonists Vanilloids specifically act
`activation and then the desensitization of
`
`subset of
`
`primary afferent nerves
`involved in the genesis of
`neurogenic inflammation.7 Notably their acute pro-
`is due to the stimulation of the
`inflammatory effect
`from peripheral
`release of sensory neuropeptides
`axons of these nerves In subsequent
`phase there is
`desensitization of these fibres and if the dose and the
`duration of exposure to the agonists are appropriate
`depletion of
`the neuropeptide
`these
`content of
`nerves occurs.8 Desensitization andor neuropeptide
`determines
`an anti-inflammatory
`effect
`depletion
`blocking the induction of neurogenic inflammation
`by endogenous or exogenous inflammatory stimuli.7
`
`2000 Taylor
`
`Francis Ltd
`
`223
`
`FAMY CARE - EXHIBIT 1019-0001
`
`

`

`Vieira et al
`
`that we found that RA is
`Noteworthy
`is the fact
`devoid of the pungent properties typical of capsaicin
`and that
`it does not exert any hyperalgesic effect
`towards heat and chemical nociceptive stimuli Vieira
`These similarities and the
`ref
`et cii submitted
`differences lack of pungent properties with capsai
`render RA
`promising analgesic and anti-inflam
`cm
`local application
`Long-term
`matory
`via its
`agent
`topical capsaicin application in rats has been shown
`to produce
`reversible impairment of
`the primary
`afferent
`fibres in the skin.0
`In view of the above we have assessed the effect of
`acute or repetitive RA topical
`in several
`treatment
`models of both acute and subchronic inflammation
`
`Materials and methods
`
`Animals
`
`Male albino Dunkin-Hartley guinea-pigs 250-350
`from Charles River Calco Italy and male Swiss mice
`2025
`from Harlan Nossan Correzzana Milan
`Italy were used The animals were fed with
`standani
`diet and water ad libitum The experiments were
`performed in respect of the Frinciples of Laboratory
`Animal Care NIH publication no 85-23 revised
`1985 and in accordance with the Italian Health
`the care and use of experi
`
`Ministry guidelines for
`mental animals
`
`Oedema induction in the paw of mice
`
`Paw oedema
`
`i.d
`was
`induced
`by intradermal
`right paw of 300 ig/0.04 ml of
`in the
`injection
`carrageenan The thickness of the paw was measured
`in mm using micro-calibrator
`In acute experiments
`the paw thickness was measured before and
`and
`injection In chronic
`after the carrageenan
`experiments the progression of oedema was assessed
`on the first day and the eighth day each hour during
`the 6-h period following the first or second carragee
`nan injection respectively Single or repeated
`days
`RA
`of vehicle peanut oil
`applications
`topical
`0.09 mg/mouse
`0.9 mg/mouse
`or
`at
`capsaicin
`in preliminary experiments were
`chosen
`doses
`applied on the ventral surface of the right paw The
`first and the last application were made 30 min before
`the injection of carrageenan
`In other experiments the repeated topical applica
`days of vehicle peanut oil RA 0.9 mg/
`tion
`mouse or capsaicin 0.09 mg/mouse or the repeated
`days i.d administration of RA potassium salt
`on
`0.03 mg/mouse or capsaicin 0.003 mg/mouse
`the ventral surface of the right paw was tested in the
`adjuvant 30 il
`of complete Freunds
`presence
`injected in the hind-paw on the first day The paw
`week for
`weeks
`volume was monitored once
`as
`described above
`
`224
`
`Mediators of Inflammation
`
`Vol
`
`2000
`
`Eyelid oedema induction in guinea-pigs
`
`oedema was
`induced
`by i.d injection
`of
`Eyelid
`10 ig/0 ml of histamine in the right superior eyelid
`of animals under
`short anaesthesia by diethyl ether
`The thickness of the eyelid was measured in mm using
`ophthalmic micro-callipers Dixey UK In acute
`experiments the eyelid thickness was measured
`and 6h after the histamine
`before and
`injection or maximal applicable doses of RA 100 mg/
`guinea-pig and capsaicin 10 mg/guinea-pig
`In subchronic experiments the animals were
`or 22 days with vehicle peanut
`treated topically for
`oil RA 0.9 mg/guinea-pig
`or capsaicin 0.09 mg/
`guinea-pig and the progression of the oedema was
`assessed on the first day during
`following the first
`histamine injection and then the eighth and the 22nd
`the second
`day each hour
`6-h period after
`for
`injection of histamine
`
`Radioimmunoassay RIA
`
`at
`
`For the tissue peptide measurements the paws were
`The
`rapidly removed and weighed
`samples were
`acid at 95C for
`extracted with ml of
`acetic
`15 min They were then homogenized and centrifuged
`l0000g for 3Omin at 4C the supernatants were
`freeze-dried and stored at 20C until their content of
`substance P-like immunoreactivity SP-LI was deter
`mined by RIA Lyophilized samples were reconstituted
`in lOml of 50 mM phosphate buffer pH 7.4
`containing 0.3% bovine serum albumin and 10 mM
`ethylene diamine tetraacetic acid EDTA The RIA
`assay for SP-LI was based on scintillation proximity
`as previously described The
`assay technology
`incubation mixture was composed of 100 pd of the
`sample diluted 130 100 pd
`of
`reconstituted
`251-labelled tracer about 6000 cpm 100 pd of diluted
`antiserum 1100 000 for SP and 50 pd of scintillation
`reagent After an overnight
`proximity assay protein
`incubation at room temperature under gentle agi
`tation the samples were counted in
`3-scintillation
`counter 2200CA Canberra-PackardUSA
`
`Chemicals
`
`RA 99% pure Sigma was diluted with peanut oil
`Sigma its soluble potassium salt MEN 11938 or
`from
`potassium
`-1 2-hydroxy- 9-octadecenoate
`Menarini Ricerche
`spa
`Chemistry Department
`Pomezia Italy was used for i.d treatment dissolved in
`saline Capsaicin Serva was dissolved in 10% etha
`nol 10% tween 80 and 80% saline
`solution and
`applied locally in peanut oil Histamine Sigma was
`type II Sigma in
`dissolved in saline carrageenan
`sterile phosphate buffer solution PBS Sigma Com
`plete Freunds adjuvant was purchased from Sigma
`Bolton Hunter SP and scintillation proximity
`
`FAMY CARE - EXHIBIT 1019-0002
`
`

`

`Anti-inflammatory activity of ricinoleic acid
`
`1.2
`
`0.8
`
`0.6
`
`0.4
`
`Time
`
`Carrageenan-induced oedema of mouse paw Effects
`FIG
`of 8-day treatment with ricinoleic acid RA 0.9 mg/mouse or
`capsaicin Cps 0.09 mg/mouse Data are mean
`standard
`error SE
`68 for each group
`0.01
`versus vehicle group
`
`0.001
`
`of treatment capsaicin 0.09 mg/mouse produced
`reaction that disappeared within
`slight hyperemic
`12 from treatment On the eighth day the carragee
`nan-induced oedema was virtually abolished in RA or
`Thus the mice
`capsaicin pre-treated animals Fig
`were unable to produce any oedema in response to
`carrageenan
`
`Effect of topical and i.d treatment with RA or
`capsaicin on Freunds adjuvant-induced paw
`oedema in mice
`
`The intraplantar
`
`treatment
`
`injection of Freund adjuvant in one
`inflam
`of the hind-paws leads to localized unilateral
`the paw oedema induced
`mation.As shown inTable
`weeks after
`by Freunds adjuvant peaked at the first
`the induction and then declined Topical
`with RA for
`days was effective
`in reducing the
`oedema formation at week observation time On the
`second and third weeks after the oedema induction
`RA did not affect
`the increase in thickness of the paws
`as compared with the vehicle-treated group Capsai
`induce significant
`cin in the applied dose did not
`inhibition in the oedema evoked by Freund adjuvant
`Table
`Conversely the i.d administration of the potassium
`salt of RA MEN 11938 was able to exert marked
`that lasted for weeks Table
`anti-inflammatory effect
`The values of reduction in oedema formation were
`68 67 and 85% of control at
`and
`weeks after
`Freunds adjuvant
`injection Similarly i.d capsaicin
`oedema
`formation the sig
`affected
`weeks the
`nificance of the effect
`lasting only for
`oedema was reduced by 50 57 and 51% at
`and
`weeks after its induction Table
`The i.d injection
`of capsaicin 0.003 mg/mouse but not RA 0.03 mg/
`mouse produced nociceptive reactions during the
`first 23 days of treatment
`
`significantly
`
`Mediators of Inflammation
`
`Vol
`
`2000
`
`225
`
`assay reagents were obtained from Amersham The
`rabbit anti-SP serum RAS 7451 cross-reactivity
`5%
`with SP 7-11 0.01% with neurokininA NKA was
`from Peninsula Laboratories
`
`Statistical analysis
`
`The data are expressed as means
`standard error of the
`mean SEM The
`significance between
`statistical
`groups was assessed using one-way analysis of variance
`followed by Bonferronis test The Mann-Whitney
`test was used for the in vitro experiments
`
`Results
`
`Effects of acute and subchronic topical
`treatment with RA and capsaicin on
`carrageenan-induced paw oedema in mice
`
`its
`
`still
`
`paw oedema induced
`As shown in Fig
`by
`carrageenan was markedly increased
`by the topical
`of RA 0.9 mg/mouse
`or
`capsaicin
`application
`0.09 mg/mouse The enhancement by RA or capsai
`oedema
`reached
`of the carrageenan-induced
`cm
`maximum between the second and fourth hours and
`then decreased
`remained sig
`time but
`over
`the sixth hour
`as compared with the
`nificant
`at
`No enhancement of
`vehicle-treated paws Fig
`oedema was detected with the
`carrageenan-induced
`or with
`solution at the same pH of
`vehicle
`100mg of RA
`Topical administration of
`vehicle RA up to 10 mg/b
`pd or capsaicin
`up to
`pd did not produceper se any oedema in the
`mg/b
`paws of the mice
`The treatment with RA 0.9 mg/mouse for
`days
`result per se in any oedematous effect or
`did not
`hyperemic reaction Conversely during the first days
`
`Vehicle
`-U- RA
`0-- Cps
`
`1.6
`
`F-
`
`1.2
`
`JO.8
`
`Time
`Effects of vehicle ricinoleic acid RA 0.9 mg/mouse
`FIG
`or capsaicin Cps 0.09 mg/mouse on carrageenan-induced
`paw oedema expressed in mm Ricinoleic acid capsaicin
`and
`the vehicle were applied topically 30 mm before carrageenan
`standard error SE
`68 for
`injection Data are mean
`each group
`0.001 versus vehicle group
`0.01
`
`FAMY CARE - EXHIBIT 1019-0003
`
`

`

`Vieira et al
`
`in mice Animals were treated by local application of ricinoleic acid RA
`Paw oedema induced
`by Freunds adjuvant
`Table
`days with MEN 11938 30 pg/mouse
`900 pg/mouse or capsaicin 90 pg/mouse for
`days or intradermally administered for
`the potassium salt of RA or capsaicin
`pg/mouse The paw oedema was measured once weekafterthe
`injection of Freunds
`adjuvant
`
`Local application for
`Vehicle
`RA 900pgmouse
`Capsaicin 90pgmouse
`
`days
`
`Intradermal
`
`treatment
`
`for
`
`days
`
`Vehicle
`MEN 11938 30pgmouse
`Capsaicin 3pgmouse
`
`First week
`
`Second week
`
`Third week
`
`1.39
`0.10
`0.82 0.10
`0.87
`0.13
`
`1.19
`0.09
`0.38 0.10
`0.69 0.09
`
`0.79
`
`0.61
`
`0.70
`
`1.20
`
`0.40
`
`0.61
`
`0.06
`
`0.13
`
`0.08
`
`0.12
`0.13
`0.08
`
`0.60
`
`0.40
`
`0.48
`
`0.63
`
`0.09
`
`0.31
`
`0.10
`
`0.07
`
`0.09
`
`0.10
`0.13
`0.16
`
`Data are mean
`
`standard error SE
`
`810 for each group
`
`0.05
`
`0.01 versus respective
`
`vehicle group
`
`Effect of topical application of RA or capsaicin
`on guinea-pig eyelid
`
`The topical administration of capsaicin
`resulted in
`local hyperemic response and activation of behav
`including scratching of the
`ioural defensive responses
`eyelid Neither hyperemia nor aversive behaviour was
`observed following RA topical
`treatment
`eyelid oedema with the peak at
`following applica
`tion with high doses of RA 100 mg/guinea-pig 0.23
`0.01 mm or capsaicin
`10 mg/guinea-pig
`0.35
`0.005 mm alone was observed
`
`slight
`
`Effects of acute and subchronic topical
`treatment with RA and capsaicin on eyelid
`oedema induced by histamine in guinea-pig
`
`The injection of histamine in the right eyelid rapidly
`an oedema
`the maximum
`induced
`reached
`that
`During id injection
`thickness at the first hour Fig
`
`Vehicle a- RA
`
`Cps
`
`Time
`
`oedema of guinea-pig eyelid
`FIG
`Histamine-induced
`Effect of vehicle ricinoleic acid RA 0.9 mg/guinea-pig
`and
`capsaicin Cps 0.09 mg/guinea-pig
`acid capsai
`and the
`vehicle were administered
`30 mm
`topically
`before histamine injection Data are mean
`standard error
`SE
`58 for each group
`0.05
`0.01
`0.001 versus vehicle group
`
`Ricinoleic
`
`cm
`
`226
`
`Mediators of Inflammation
`
`Vol
`
`2000
`
`for
`
`effect
`
`the animals showed aversive response behaviour but
`no scratching response on the eyelid was usually
`oedema was
`observed As shown in Fig
`the
`by the topical application of RA 0.9 mg/
`increased
`guinea-pig and capsaicin 0.09 mg/guinea-pig
`The topical application of RA 0.9 mg/guinea-pig
`result per se in any oedematous
`days did not
`reaction
`or hyperemic
`Instead
`capsaicin
`produced hyperemic and pain
`0.09 mg/guinea-pig
`reactions that disappeared 12
`after the treat
`ful
`ment The
`these
`intensity of
`gradually
`responses
`starting from 23 days and was almost
`decreased
`abolished at the end of the treatment As shown in Fig
`the eyelid oedema
`induced
`by histamine was
`markedly reduced after
`days of
`treatment
`topical
`with RA 0.9mg/guinea-pig or capsaicin 0.O9mg/
`similar profile of oedema reduction was
`guinea-pig
`treatment with RA
`also found after
`weeks of topical
`
`0.09 mg/guinea-
`0.9 mg/guinea-pig
`or
`capsaicin
`pig the values of eyelid oedema at
`the peak first
`hour being 1.37
`0.05 and 0.85
`0.06 0.70
`
`Vehicle ci--- RA
`
`Cps
`
`1.5
`
`105
`
`Time
`
`Histamine-induced oedema of guinea-pig eyelid
`FIG
`Effect of 8-day treatment with ricinoleic acid RA 0.9 mg/
`guinea-pig or capsaicin Cps 0.09 mg/guinea-pig
`Data are
`standard error SE
`58 for each group
`0.05
`mean
`0.001 versus vehicle group
`0.01
`
`FAMY CARE - EXHIBIT 1019-0004
`
`

`

`0.05 mm for
`groups respectively
`
`vehicle-
`
`RA-
`
`and
`
`capsaicin-treated
`
`ceptive reactions observed during the first days only
`
`Anti-inflammatory activity of ricinoleic acid
`
`disap
`i.d injection
`application or
`after capsaicin
`peared on the second or third day of treatment
`known that repeated exposure to capsaicin results in
`desensitization of chemosensitive sensory affer
`ents8 depletion of sensory peptides such as
`and
`tissue reactions upon chemical
`loss of local
`flare reaction induced
`irritation.20 Desensitization of
`
`is
`
`It
`
`i.d
`
`by
`
`prolonged local exposure to capsaicin
`and histamine has been
`described in
`bradykinin
`human skin.2 The anti-inflammatory effect of RA and
`found in our experimental conditions was
`capsaicin
`than histamine
`more marked towards carrageenan
`different mechanisms of action of these inflammatory
`agents can be responsible for these differences in
`desensitization Histamine has been
`described to
`
`Effect of repeated treatment of RA and
`capsaicin on mouse paw SP levels
`
`The levels of SP were not different
`in carrageenan
`0.28 and
`inflamed and non-inflamed paws 1.30
`0.34 pmol/g tissue The repeated
`days local
`1.09
`administration of RA at doses able to produce marked
`anti-inflammatory effects see above reduced the SP
`content by 58% in the inflamed paw of mice 0.54
`0.05 as compared with the
`0.12 pmol/g tissue
`inflamed paw treated with vehicle Repeated topical
`capsaicin administration did not significantly reduce
`SP paw levels 0.73
`0.14 pmol/g tissue
`
`Discussion
`
`administered for
`
`matory effect
`
`this study is that RA topically
`The main result of
`days exerts marked anti-inflam
`in several models of inflammation The
`effect was comparable with that
`anti-inflammatory
`10-fold lower dose of capsaicin
`obtained with
`RA when administered acutely produced an addi
`tive inflammatory effect on carrageenan- or histamine-
`induced oedema in mice or guinea-pigs respectively
`variance with capsaicin acute topical RA
`but at
`administration did not produce any nociceptive or
`hyperemic reaction Capsaicin is known to induce
`neurogenic inflammation e.g vasodilatation protein
`extravasation and oedema through an acute release of
`and
`such as SP neurokinin
`peptides
`vasoactive
`peptide CGRP from sensory
`calcitonin gene-related
`nerve terminals.2 -14
`is noteworthy that capsaicin
`induced oedema or plasma protein extravasation is
`almost completely absent
`in genetically tachykinin
`deficient mice.5
`release oftachykinins from sensory
`leading to an activation of
`their
`terminal nerves
`receptors present on the skin surface may explain the
`additive inflammatory effect of RA in our
`acute
`experimentally induced oedemas
`Although carrageenan
`injection leads to the genera
`inflammatory mediators the
`tion of
`of
`oedema
`of paw
`thickness
`noticeable
`reduction
`induced by the NK receptor antagonist CP 96.345 in
`rats6 and the almost complete disappearance
`of paw
`oedema in mice induced by the other NK receptor
`antagonist CP 122721 Vieira et cii unpublished
`the involvement of SP in this
`observations
`support
`inflammatory oedema Furthermore in NK receptor
`oedema was sig
`knockout mice the carrageenan
`nificantly reduced.7
`As concerns the anti-inflammatory
`resulting from an 8-day treatment
`desensitization
`action on peripheral nerve endings already described
`in part such
`for capsaicin may explain at
`least
`In keeping with the above the noci
`phenomenon
`
`It
`
`range
`
`effect of RA
`
`excite small diameter afferent neurones and evoke the
`release of neuropeptides from vasoactive local nerve
`is interesting to note that paw oedema
`endings.22 It
`induced by histamine was abolished in capsaicin
`denervated rats.23 Our
`findings in the guinea-pig
`the histamine-induced oedema
`eyelid show that
`also dependent on other mediators
`On the other hand carrageenan
`paw oedema was
`blunted by the local application of RA or capsaicin and
`reduction of SP tissue
`it was matched by
`significant
`levels at least after the local application of RA
`In contrast to what happens in acute inflammation
`in subchronic
`inflammation induced by Freund
`adjuvant RA was barely more effective than capsaicin
`in reducing oedema formation Freunds adjuvant
`relatively stable inflammation that
`able to produce
`was decreased partially by local application of RA and
`was markedly affected by i.d injection of both RA and
`capsaicin The more effective
`counteraction of the
`oedema induced by i.d versus topical administration
`of both compounds is probably due to
`stronger
`desensitization induced by this route On the other
`hand Freunds adjuvant-induced chronically inflamed
`tissue in the rat has been reported to be associated
`with changes in SP content
`in nerves supplying the
`inflamed paws24 and systemic capsaicin administra
`tion reduced the oedema
`formation concomitantly
`with the reduction of sp in nerve tissues.25
`the data presented should be
`The
`relevance
`of
`associated with the observation that RA does not
`possess the pungent and painful effect of capsaicin
`but maintains its anti-inflammatory activities Topical
`has shown therapeutic
`cutaneous
`disorders
`of
`
`capsaicin
`
`treatment
`
`potential
`such
`
`is
`
`is
`
`in the
`
`as post
`herpetic neuralgia painful diabetic neuropathy pruri
`tus psoriasis post-mastectomy pain syndrome vulvar
`vestibulitis 26-29 but
`to be limited
`appears
`its utility
`properties RA has
`primarily by its
`irritant
`new capsaicin-like
`be
`substance
`to
`potential
`endowed with anti-inflammatory
`effects on several
`models of inflammation without the pungent charac
`teristics of capsaicin
`
`the
`
`Mediators of Inflammation
`
`Vol
`
`2000
`
`227
`
`FAMY CARE - EXHIBIT 1019-0005
`
`

`

`an in vivo
`Immunol
`
`to bradykinin
`
`Lundberg
`
`Tyers MR The non-peptide
`16 Birch PJ Hanison SM Hayes AG Rogers
`-CP-96345 produces antinociceptive
`and
`NK-1 receptor antagonist
`1992105508-10
`anti-oedema effects in the rat BrJ Pharmacol
`HoultJR Brain SD Neurokinin-1
`17 CaoT PinterEAl-RashedS Gerard
`receptor agonists are involved in mediating neutrophil
`accumulation in
`the inflamed but not normal cutaneous microvasculature
`knockout mice
`neurokinin -1
`study using
`receptor
`20001645424-9
`18 Jancsô
`Direct evidence for neurogenic
`Jancsô-GâborA
`Szolcsányi
`by denervation
`and by pre-treatment
`and
`inflammation
`its prevention
`with capsaicin BrJ Pharmacol Chemother 196731138-51
`19 JesselTM Iversen LL Cuello AC Capsaicin induced depletion of SP from
`primary sensory neurons Brain Res 1978152183-8
`20 Gamse
`Holzer
`Lembeck
`Decrease of SP in primary afferent
`neurons and impairment of neurogenic plasma extravasation by capsai
`cm BrJ Pharmacol
`198068207-13
`21 Crimi
`Polosa
`Maccarone
`Palermo MistrettaA Effect of topical
`application with capsaicin
`and
`on skin responses
`histamine in man Gun Etp Allergy 199222933-9
`22 SariaA Martling C-RYan
`Theodorsson-Norheim
`Gamse
`JM Release
`from capsaicin-sensitive
`of multiple tachykinins
`sensory
`nerves in the lung by bradykinin
`histamine dimethylphenyl
`pipcr
`nerve stimulation Am Rev Respir Dis 1988137
`azinum and
`13 305
`23 Amaim Schuligoi
`Lanz
`Doimerer
`edema in
`Histamine-induced
`CGRP receptor
`the rat paw
`effect of capsaicin denervation
`1995279227-31
`antagonist EurJ Pharmacol
`24 Donnerer
`and transport
`Increased content
`Stein
`of
`Schuligoi
`and CGRP in sensory nerves iimervating inflamed tissue
`substance
`evidence for
`function
`in viva
`of nerve growth
`factor
`regulatory
`Neuroscience 199249693-8
`25 Colpaert FC DonnererJ
`Effect of capsaicin on inflammation
`Lembeck
`and on the substance
`content of nervous tissues in rats with adjuvant
`Life Sc 1983321827-34
`26 Bernstein JE Parish LC RapaportM Rosenbaum MM Rocni HH Jr
`topically applied capsaicin on moderate and severe psoriasis
`vulgaris JAm AcadDermatol 198615504-7
`27 Bernstein JE Korman NJ Bickers DR Dahl
`of chronic
`capsaicin treatment
`Dermatol 198921265-70
`28 Carter RB Topical capsaicin in the treatment of cutaneous disorders
`Drug Dcv Res 199122109-23
`29 Watson CPN Evans RJ The post-mastectomy pain syndrome and topical
`randomised trial Pain 199251375-9
`capsaicin
`
`vagal
`
`and
`
`arthritis
`
`Effect of
`
`post-herpetic
`
`Milikan LE Topical
`Am Acad
`
`neuralgia
`
`Received
`
`accepted
`
`August 2000
`September 2000
`
`Vieira et al
`
`We would like to thank Dr it Terraeeiano for the
`ACKNOWLEDGEMFThff
`and synthesis of MEN 11938 Mrs
`Mylander
`development
`the English text and Ms
`Azzurrini for her secretarial assistance
`and Dr
`Olivein from the Faculty of Medical Sciences
`of Ribeirao Preto USP Bnzil
`and CNPq Brazil
`
`for revision of
`
`References
`
`StewartJJ Gaginella TS Olsen WA Bass
`on motility and water and electrolyte
`transport
`tract.JPharmacol Etp Ther 1975192458-67
`mechanisms of
`Dny
`Neuropharmacological
`199244611-5
`substances Bloc/acm
`Pharmacol
`The vanifloid capsaicin receptor receptor
`Szàllasi
`differences en Pharmacol
`199425223-43
`Szabo
`et at
`triprenyl phenol of fungal
`non-pungent
`stimulates nit dorsal ganglion neurons via intenction
`origin scutigeral
`at vanifloid receptors BrJPharmacol
`19991261351-8
`Klopman
`activity relationship of
`Li J-Y Quantitative
`structure-agonist
`Comp Alt MOI Des 19959283-94
`capsaicin analogues
`Caterina MJ Shumaker MA TominagaM RosenTA LevineJD Julis
`heat-activated ion channel
`capsaicin receptor
`in the pain pathway
`Nature 1997389816-24
`Maggi CA Mcli
`The sensory-efferent
`function
`sensory neurons Gen Pharmacol
`1988191-43
`Holzer
`targets mechanisms
`Capsaicin
`neurons
`Pharmacol
`
`Szàllasi
`
`Biro
`
`cellular
`
`for
`
`thin sensory
`
`selectivity
`
`143 20
`
`Inhibitory
`
`actions of laxatives
`
`in the
`
`gastrointestinal
`
`capsaicin and
`
`related
`
`type and species
`
`The
`
`of capsaicin-sensitiw
`
`of
`
`and
`action
`199143
`Rev
`
`Vieira
`
`Evangelista
`
`Cirillo RTerraciano
`
`Santicioli
`
`long-term
`
`LippiA Maggi CA Manzini
`capsaicin-like compound
`acid
`Antinociceptive
`activity of ricinoleic
`2000407109-16
`devoid of pungent properties Eur Pharmacol
`10 McMahon SB Lewin
`Bloom SR The consequences of
`in the rat Pain 199144301-10
`topical capsaicin application
`CriscuoliM Maggi CA Depolarisation
`evoked co
`11 Lippi
`from enteric nerves in the guinea-pig proximal
`release of
`tachykinins
`bergs Arc Pharmacol 1998357245-51
`colon Naunyn-Schniede
`12 Jancsô
`The role of sensory nerve endings
`Jancsô-GâborA Szolcsányi
`inflammation induced in human skin and in eye and paw
`in neurogenic
`-41
`19683232
`of the rat BrJ Pharmacol Chemother
`Lundberg JM Anggard
`13 Lundblad
`Increased vascular
`Saria
`in rat nasal mucosa induced
`and stimulation
`by SP
`of
`Acta Otolaryngol 198396
`neurons
`
`permeability
`
`trigeminal
`
`capsaicin-sensitive
`47 9-84
`14 SariaA LundbergJM Skofitsch
`Lembeck
`in
`Vascular protein leakage
`various tissues induced by S1 capsaicin bradykinin serotonin histamine
`Arch Pharmacol
`and by antigen challenge Naunyn-Schmiedebergs
`-8
`1983324212
`15 Cao QY Mantyh
`Carison EJ Gillespie A-M Epstein CJ BasbaumAl
`are required to experience moderate to
`Primary afferent
`tachykinins
`intense pain Nature 1998392390-4
`
`228
`
`Mediators of Inflammation
`
`Vol
`
`2000
`
`FAMY CARE - EXHIBIT 1019-0006
`
`