[Logo: VIVACY
`LABORATORIES]
`
`Opposition to the patent
`EP2 349 203 B1 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`ANNEX 7A
`
`RA1336402
`
`Version A
`
`Page 1 of6
`
`1) Objective - Aim
`
`These tests were conducted by Vivacy within the framework of of a project. They are reused as part of the
`argument provided in the notice of opposition filed against the issuing of patent EP 2 349 203.
`
`2) Protocol - Method
`
`a. Manufacture of gels
`
`Cross-linked hyaluronic acid gels
`
`Gels containing cross-linked hyaluronic acid are obtained according to the operating method described in the
`patent application WO 2009/071697 in the name of the requester from sodium hyaluronate fibers (NaHa) and
`butanediol diglycidyl ether (BODE). The final gels have a hyaluronic acid concentration of around 20 mg/g.
`
`The conditions for cross-linking are as follows: 50.5°C - 2h 45min. The cross-linking ratio is defined as the
`relationship between the number of moles of cross-linking agent introduced into the reaction medium and the
`number of moles of disaccharide motifs introduced into the reaction medium.
`
`Several cross-linking ratios were deployed: X=0.06, X=0.07 and X=0.12.
`
`Several molecular weights of hyaluronicacid were deployed: 1 MDa and 3 MDa.
`
`Non-cross-linked hyaluronic acid gels
`
`High-quality injectable sodium hyaluronate (NaHa) fibers are weighed in a receptacle. An aqueous phosphate
`buffer solution is added, the whole is homogenized for around 1 hour with a spatula, at ambient temperature and
`under atmospheric pressure of 900 mmHg. The final gels have a hyaluronic acid concentration of around 20
`mg/g.
`
`Just one molecular weight of hyaluronic acid was deployed: 3 MDa.
`
`Lidocaine
`
`Lidocaine is prepared in a phosphate buffer solution at pH levels close to 7.
`
`Additional compounds (magnesium ascorbyl phosphate, sucrose octasulfate)
`
`The additional compounds are solubilized in a phosphate buffer solution before being incorporated in hyaluronic
`acid gels.
`
`Sterilization
`
`The formulations thus obtained are packaged in syringes which are sterilized by autoclaving in steam (T=121 °C,
`10 min).
`
`RDDC004 version A
`
`© Reproduction and distribution prohibited without authorization by VIVACY®
`
`ALL 2084
`PROLLENIUM V. ALLERGAN
`IPR2019-01505 et al.
`
`

`

`[Logo: VIVACY
`LABORATORIES]
`
`Opposition to the patent
`EP2 349 203 B1 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`b. Measurement of rheological properties
`
`ANNEX 7A
`
`RA1336402
`Version A
`
`Page 2 of 6
`
`The elastic G' components of formulations containing cross-linked hyaluronic acid before and after sterilization
`by steam autoclaving were measured on a TA Instruments AR 2000ex rheometer, oscillating at 25°C, the values
`for the elastic G' component being noted at a frequency of 1 Hz.
`
`The viscous components were also measured.
`
`The viscosity ri of formulations containing non-cross-linked hyaluronic acid is measured on a TA Instruments AR
`2000ex rheometer, under imposed stress at 25°C. The viscosity value is noted at a stress of 0.02 s·1.
`
`3) Results
`
`The results below illustrate the influence of lidocaine in the presence of mannitol on the degradation during heat
`sterilization of the rheological properties of a hyaluronic acid gel with a weight-average molecular weight of 3
`MOa with a cross-linking ratio X=0.12 at a concentration of 20 mg/g.
`
`At 0.1 % lidocaine there is no detrimental effect, mannitol succeeds in compensating for the effects of lidocaine.
`Conversely from 0.3% upwards the beneficial effects of mannitol are destroyed by lidocaine.
`
`Sterilization
`121 °C-10 min
`
`3.5% Mannitol 1 %
`Lidocaine
`3.5% Mannitol 0.6%
`Lidocaine
`3.5% Mannitol 0.3%
`Lidocaine
`3.5% Mannitol 0.1%
`Lidocaine
`3.5% Mannitol
`8.5 g/1 NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G' (1 Hz)
`G" (1Hz)
`G' (1 Hz)
`G" (1Hz)
`252
`31.8
`141
`22.4
`
`Tan o
`0.1265
`
`Tan o
`0.1593
`
`Variation
`G' (1 Hz)
`-44%
`
`258
`
`252
`
`256
`
`254
`221
`
`32.3
`
`31.4
`
`32.0
`
`32.0
`27.3
`
`0.1252
`
`151
`
`0.1246
`
`175
`
`23.2
`
`25.4
`
`0.1535
`
`-41%
`
`0.1450
`
`-31 %
`
`0.1256
`
`205
`
`27.25
`
`0.1329
`
`-20%
`
`0.1256
`0.1233
`Table 1
`
`197
`140
`
`27.4
`21 .1
`
`0.1391
`0.1511
`
`-22%
`-37%
`
`RDDC004 version A
`
`©Reproduction and distribution prohibited without authorization by VIVACY®
`
`

`

`[Logo: VIVACY
`LABORATORIES)
`
`Opposition to the patent
`EP2 349 203 81 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`ANNEX 7A
`
`RA1336402
`
`Version A
`
`Page 3 of 6
`
`The results below illustrate the influence of lidocaine in the presence of mannitol on the degradation during heat
`sterilization of the rheological properties of a hyaluronic acid gel with a weight-average molecular weight of 1
`MDa at a concentration of 20 mg/g, with a cross-linking ratio X= 0.07.
`
`Whatever the concentrations of lidocaine tested, the beneficial effects of mannitol are destroyed by lidocaine.
`
`Sterilization
`121 °C-10min
`
`3.5% Mannitol 0.6%
`Lidocaine
`3.5% Mannitol 0.3%
`Lidocaine
`3.5% Mannitol
`8.5 g/I NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G' (1 Hz) G" (1Hz)
`G' (1Hz) G" (1 Hz)
`437
`55.8
`35.4
`
`Tan o
`0. 1276
`
`288
`
`Tan o
`0.1248
`
`Variation
`G'(1 Hz)
`-34%
`
`437
`
`459
`422
`
`: 56.2
`
`0.1276
`
`315
`
`155.2
`51.8
`
`0.1202
`0.1227
`Table II
`
`373
`286
`
`41.4
`
`47.3
`33.3
`
`0.1307
`
`-28%
`
`0.1268
`0.1166
`
`-19%
`-32%
`
`The results below illustrate the influence of lidocaine in the presence of mannitol on the degradation during heat
`sterilization of the rheological properties of a hyaluronic acid gel with a weight-average molecular weight of 3
`MDa at a concentration of 20 mg/g with a cross-linking ratio X=0.06.
`
`Whatever the concentrations of lidocaine tested, the presence of lidocaine destroys the effects of mannitol.
`
`Sterilization
`121 °C-10 min
`
`3.5% Mannitol 0.3%
`Lidocaine
`3.5% Mannitol
`8.5 g/I NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G' (1 Hz)
`G" (1Hz)
`G' (1 Hz)
`G" (1 Hz)
`149
`99
`
`Tana
`0.1785
`
`26.6
`
`Tan o
`0.2278
`
`Variation
`G' (1 Hz)
`-34%
`
`22.6
`
`147
`157
`
`26.3
`25.5
`
`0.1794
`0.1628
`Table Ill
`
`129
`114
`
`25.1
`22.4
`
`0.1955
`0.1968
`
`-12%
`-27%
`
`RDDC004 version A
`
`0Reproduction and distribut ion prohibited w it hout authorization by VIVACY®
`
`

`

`[Logo: VIVACY
`LABORATORIES]
`
`Opposition to the patent
`EP2 349 203 B1 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`ANNEX 7A
`
`RA1336402
`
`Version A
`
`Page 4 of 6
`
`The results below illustrate the influence of lidocaine in the presence of mannitol and of SOS on the degradation
`during heat sterilization of the rheological properties of hyaluronic acid gels with a weight-average molecular
`weight of 3 MDa with a cross-linking ratio X=0.12 at a concentration of 20mg/g.
`
`In the presence of SOS the effects of mannitol are destroyed by lidocaine at a concentration of 0.3%,
`
`Sterilization
`121 °C- 10 min
`
`0. 1 % Sucrose
`octasulfate 3.5%
`Mannitol 0.3%
`Lidocaine
`0.1 % Sucrose
`octasulfate 3.5%
`Mannitol
`8.5 g/1 NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G'(1Hz)
`G" (1 Hz)
`G' (1 Hz) G" (1 Hz)
`256
`31 .55
`187
`25.88
`
`Tan o
`0.1233
`
`Tan o
`0.1389
`
`Variation
`G'(1Hz)
`-27%
`
`254
`
`31.31
`
`0.1234
`
`197
`
`26.6
`
`0.1350
`
`-22%
`
`221
`
`27.3
`
`0.1233
`Table IV
`
`140
`
`21.1
`
`0.1511
`
`-37%
`
`The results below illustrate the influence of lidocaine in the presence of mannitol and of MAP at a concentration
`of 0.3 mg/g, on the degradation during heat sterilization of the rheological properties of hyaluronic acid gels with
`a weight-average molecular weight of 3 MDa with a cross-linking ratio X=0.12 at a concentration of 20 mg/g.
`
`In the presence of MAP, the effects of mannitol are destroyed by lidocaine at a concentration of 0.3%.
`
`Sterilization
`121°C-10 min
`
`0.03% Magnesium
`ascorbyl phosphate
`3.5% Mannitol 0.3%
`Lidocaine
`0.03% Magnesium
`ascorbyl phosphate
`3.5% Mannitol
`8.5 g/I NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G' (1Hz)
`G" (1Hz)
`G' (1 Hz) G" (1Hz)
`264
`38.39
`187
`31.93
`
`Tan o
`0.1454
`
`Tan o
`0.1708
`
`Variation
`G'(1 Hz)
`-30%
`
`270
`
`38.6
`
`0.1431
`
`209
`
`33.63
`
`0.1607
`
`-23%
`
`221
`
`27.3
`
`0.1233
`Table V
`
`140
`
`21 .1
`
`0.1511
`
`-37%
`
`RDDC004 version A
`
`©Reproduction and distribution prohibited without authorization by VIVACY®
`
`

`

`(Logo: VIVACY
`LABORATORIES]
`
`Opposition to the patent
`EP2 349 203 8 1 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`ANNEX 7A
`
`RA1336402
`
`Version A
`
`Page 5 of6
`
`The results below illustrate the influence of lidocaine in the presence of mannitol and of MAP at a concentration
`of 0. 7 mg/g, on the degradation during heat sterilization of the rheological properties ofhyaluronic acid gels with
`a weight-average molecular weight of 3 MDa with a cross-linking ratio X=0.12 at a concentration of 20mg/g.
`
`In the presence of MAP, the effects of mannitol are destroyed by lidocaine at a concentration of 0.3%.
`
`Sterilization 121°c -
`10 min
`
`0.07%
`Magnesiumascorbyl
`phosphate 3.5%
`Mannitol
`0.03%Lidocaine
`0.07%
`Magnesiumascorbyl
`phosphate 3.5%
`Mannitol
`8.5 g/1 NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`G'
`G"
`G'
`G"
`(1Hz)
`(1Hz)
`(1 Hz)
`(1Hz)
`264
`41 .00
`190
`32.35
`
`Tan o
`
`0.1464
`
`Tan o
`
`Variation
`G' (1 Hz)
`
`0.1705
`
`-28%
`
`266
`
`38.03
`
`0.1430
`
`208
`
`33.90
`
`0.1628
`
`-22%
`
`221
`
`27.3
`
`0.1233
`Table VI
`
`140
`
`21 .1
`
`0.1511
`
`-37%
`
`The results below illustrate the influence of lidocaine in the presence of mannitol on the degradation during heat
`sterilization of the rheological properties of a hyaluronic acid gel with a weight-average molecular weight of 3
`MDa at a concentration of 20 mg/g, non cross-linked.
`
`Whatever the concentrations of lidocaine tested, the detrimental effects of this substance on the viscosity of the
`formulations lead to the systematic procurement of formulations with properties that are more degraded than
`formulations without lidocaine.
`
`Sterilization
`121°c -10 min
`
`3.5% Mannitol 1.0%
`Lidocaine
`3.5% Mannitol 0.6%
`Lidocaine
`3.5% Mannitol 0.3%
`Lidocaine
`3.5% Mannitol 0.1%
`Lidocaine
`3.5% Mannitol
`8.5 g/1 NaCl
`
`Rheological characteristics measured at 25°C
`Before sterilization
`After sterilization
`rJ (Pa.s)
`f) (Pa.s)
`1708
`
`20
`
`1670
`
`1696
`
`1660
`
`1673
`1407
`
`27
`
`59
`
`215
`
`700
`479
`
`Table VII
`
`Variation
`f)
`-99%
`
`-98%
`
`-96%
`
`-87%
`
`-58%
`-66%
`
`RDDC004 version A
`
`©Reproduction and distribution prohibited without authorization by VIVACY®
`
`

`

`[Logo: VIVACY
`LABORATORIES)
`
`Opposition to the patent
`EP2 349 203 B1 (Anteis):
`
`TESTS CONDUCTED WITHIN THE FRAMEWORK OF
`ANOTHER PROJECT
`
`ANNEX 7A
`
`RA1336402
`
`Version A
`
`Page 6 of 6
`
`4) Conclusions
`
`The presence of lidocaine (from 0.3% upwards for cross-linked gels andfrom 0.1% for non cross-linked gels)
`destroys the effects of mannitol on the rheological properties of the gel after sterilization.
`
`No formulation containing 0.3% lidocaine displays less degradation of rheological properties in comparison
`with formulations that only contain polyol.
`
`The addition of compounds such as vitamin C derivatives or sucrose derivatives does not change the effect of
`lidocaine on the rheological properties of a gel after sterilization.
`
`5) Validations
`
`Drafted by:
`Name:
`'="[il'='le~gcc:ib':'::le~)...,....,. __________ _
`On:
`_06_._01_/_20'--1_4 _ _____ ___ _
`Checked by:
`Name:
`,,:.V.:...;IT:..:..A..:.:L=-=L:...:.Y _ _ _________ _
`On:
`"'[il-'-"le=gc..:ib:..:..lec.,] ________ ___ _
`
`Post:
`Signature:
`
`"'[il=le""g""ib""le:..L] _ _ _______ _
`L.:[S::..:i.,_gn:..:..a=.:t::::u.:..:re:.il _ ________ _
`
`Post:
`Signature:
`
`"'[ilc..::le:...agc..::ib:.;,,:le:...1.)-,--________ _
`""IS~i..,_gn-'-a""t"'u""rec...] _________ _
`
`RDDC004 version A
`
`©Reproduction and distribution prohibited without authorization by VIVACY®
`
`Certified translation from French into English.
`I, Anke SEYBERTH, sworn and qualified translator for French, English and German, accredited by the French Court of Nimes, certify
`that the target language text is an accurate and complete translation of the attached document, translation registered under no.
`PP-202008-041-1 on behalf of ACSTraduction. Issued in Grenoble on August 14, 2020.
`
`

`

`v9 Y. LY. l).~.Y
`
`1) Objectlf - But
`
`Opposition a l'encontre du brevet
`EP2 349 203 B1 (Anteis) :
`ESSA/S REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`ANNEXE7A
`
`RA1336402
`Version A
`
`Page 1 sur 6
`
`Ces essais ont ete realises par VIVACY dans le cadre d'un projet. 11s sont reutilises dans le cadre de
`!'argumentation fournie dans le memolre d'opposition depose a l'encontre de la delivrance du brevet
`EP 2 349 203.
`
`2) Protocole - Methode
`
`a. Fabrication des gels
`
`Gels d'acide hyaluronique reticule
`
`Les gels comprenant de l'aclde hyaluronique reticule sont obtenus selon le mode operatoire decrit
`dans la demande de brevet WO 2009/071697 au nom de la demanderesse a partir de fibres de
`hyaluronate de sodium {NaHA) et d'ether de butanedloldlglycidyle (BODE). Les gels finaux ont une
`concentration en acide hyaluronlque d'environ 20 mg/g.
`
`Les conditions de reticulation sent les suivantes : so,s•c - 2h45. Le taux de reticulation est defini
`comme le rapport du nombre de moles de reticulant introduites dans le milieu reactionnel sur le
`nombre de moles de motif disacharidique introduites dans le milieu reactionnel.
`
`Plusleurs taux de reticulations ont ete mis en ceuvre : X=0,06, X=0,07 et X=0,12.
`
`Plusieurs masses moleculaires d'acide hyaluronique ont ete mises en ceuvre: 1 MDa et 3 MDa.
`
`Gels d'acide hyaluronique non reticule
`
`Des fibres de hyaluronate de sodium (NaHA) de qualite injectable sont pesees dans un recipient. Une
`solution aqueuse de tampon phosphate est ajoutee, le tout est homogeneise pendant environ 1
`heure a la spatule, a temperature ambiante et sous une pression atmospherique de 900 mmHg. Les
`gels finaux ont une concentration en acide hyaluronique d'environ 20 mg/g.
`
`Une seule masse moleculaire d'aclde hyaluronique a ete mise en ceuvre : 3 MDa.
`
`Lidoca'ine
`
`La lidoca'ine est preparee dans une solution de tampon phosphate a des pH proche de 7.
`
`Composes additionnels (ascorbyl phosphate de magnesium, sucrose octasulfate)
`
`Les composes additionnels sont solubilises dans une solution de tampon phosphate avant leur
`incorporation dans les gels d'acide hyaluronique.
`
`Sterilisation
`
`RDDC004 version A
`
`©Reproduction et diffusion lnterdltes sans aulortsallon de VNACY®
`
`

`

`\ly V..I.V./:>,~.Y
`
`Opposition a l'encontre du brevet
`EP2 349 203 Bl (Anteis) :
`ESSA/S REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`RA1336402
`Version A
`
`Page 2 SLIT 6
`
`b. Mesure des proprletes rheologiques
`
`Les composantes elastiques G' des compositions comprenant de l'acide hyaluronique reticule avant
`et apres sterilisation par autoclavage a la vapeur ont ete mesurees sur rheometre TA Instrument AR
`2000 Ex, en oscillation a 25°C, les valeurs de la composante elastique G' etant relevees a une
`frequence de 1 Hz.
`
`Les composantes visqueuses ont egalement ete mesurees.
`
`La viscosite ri des compositions comprenant de l'acide hyaluronique non reticule est mesuree sur
`rheometre TA Instruments AR 2000 Ex, en contrainte imposee a 2s•c. La valeur de viscosite est
`relevee a une contrainte de 0,02 s-1
`
`•
`
`3) Resultats
`
`Les resultats ci-dessous a illustrent !'influence de la lidoca'ine en presence de mannitol sur !'alteration
`lors de la sterilisation a la chaleur des proprietes rheologiques d'un gel d'acide hyaluronique de
`masse moleculaire moyenne en poids de 3 MDa avec un taux de reticulation X=0,12 a une
`concentration de 20 mg/g.
`A 0,1 % de lidoca'ine ii n'y a pas d'effet deletere, le mannitol parvient a compenser _les effets de la
`lidoca'ine. En revanche des 0,3 % les effets benefiques du mannitol sont detruits par la lidoca"ine.
`
`Sterilisation
`121 ·c -10 min
`
`3,5% Mannitol
`1 % LidocaTne
`3,5% Mannitol
`0,6% lidoca'ine
`3,5% Mannitol
`0,3% Lidoca'ine
`3,5% Mannitol
`0,1% Lidoca'ine
`3,5% Mannitol
`
`8,5 g/1 NaCl
`
`Caracteristiques rheologiques mesurees a 25°C
`Avant sterilisation
`Apres sterilisation
`G' (1Hz) G" (1Hz) Tan 6
`G' (1Hz) G" (lHz) Tan 6
`252
`31,8
`0,1265
`141
`22,4
`0,1593
`
`Variation
`G' (1Hz)
`-44%
`
`258
`
`252
`
`256
`
`254
`
`221
`
`32,3
`
`31,4
`
`32,0
`
`32,0
`
`27,3
`
`0,1252
`
`151
`
`23,2
`
`0,1535
`
`-41%
`
`0,1246
`
`175
`
`25,4
`
`0,1450
`
`-31%
`
`0,1256
`
`205
`
`27,25
`
`0,1329
`
`-20%
`
`0,1256
`
`197
`
`0,1233
`
`140
`
`27,4
`
`21,1
`
`0,1391
`
`-22%
`
`0,1511
`
`-37%
`
`Tableau I
`
`RDDC004 version A
`
`<i:lReproductlon et dillusion lnlerdiles sans autorisation de VrvACY®
`
`

`

`'vy' VLV.~.~.Y
`
`Opposition a l'encontre du brevet
`EP2 349 203 B1 (Antels) :
`ESSAJS REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`RA1336402
`Version A
`
`Page 3 SU( 6
`
`Les resultats ci-dessous illustrent !'influence de la lidoca'ine en presence de mannitol sur !'alteration
`lors de la sterilisation a la chaleur des proprietes rheologiques d'un gel d'acide hyaluronique de
`masse moleculaire moyenne en poids de 1 MDa a une concentration de 20 mg/g, avec un taux de
`reticulation de K= 0,07.
`
`Quelles que soient les concentrations de lidoca'ine testees, les effets benefiques du mannitol sont
`detruits par la lidoca'ine.
`
`Sterilisation
`121 ·c -10 min
`
`3,5% Mannitol
`0,6% Lidoca'ine
`3,5% Mannitol
`0,3% Lidoca'ine
`3,5% Mannitol
`
`8,5 g/1 NaCl
`
`Caracteristiques rheologiques mesurees a 25°C
`Avant sterilisation
`Apres sterilisation
`G' (1Hz) G" (1Hz) Tan<'i
`G' (1Hz) G" (1Hz) Tan 6
`437
`55,8
`0,1276
`288
`35,4
`0,1248
`
`Variation
`G' (lHz)
`-34%
`
`437
`
`459
`
`422
`
`56,2
`
`0,1276
`
`315
`
`55,2
`
`0,1202
`
`373
`
`41,4
`
`47,3
`
`0,1307
`
`-28%
`
`0,1268
`
`-19%
`
`51,8
`
`0,1227
`
`286
`
`33,3
`
`0,1166
`
`·32%
`
`Tableau II
`
`Les resultats ci-dessous illustrent !'influence de la lidoca'ine en presence de mannitol sur !'alteration
`lors de la sterilisation a la chaleur des proprietes rheologiques d' un gel d'acide hyaluronique de
`masse moleculaire moyenne en poids de 3 MDa a une concentration de 20 mg/g avec un taux de
`reticulation X=0,06.
`
`Quelles que soient les concentrations de lidoca'ine testees, la presence de lidoca'ine detrult les effets
`du mannitol.
`
`Caracteristiques rheoiogiques mesurees a 25°C
`Avant sterilisation
`Apres sterilisation
`G' (lHz) G" (lHz) Tan 6
`G' (lHz) G" (lHz) Tan 6
`149
`26,6
`0,1785
`99
`22,6
`0,2278
`
`Variation
`G' (1Hz)
`-34%
`
`0,1955
`
`-12%
`
`Sterilisation
`121 ·c - 10 min
`
`3,5% Mannitol
`0,3% Lidocai'ne
`3,5% Mannitol
`
`8,5 g/1 NaCl
`
`147
`
`157
`
`26,3
`
`25,5
`
`0,1794
`
`129
`
`0,1628
`
`114
`
`25,1
`
`22,4
`
`Tableau Ill
`
`0,1968
`
`-27%
`
`-
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`
`traducuon Asse(~'
`
`

`

`vy VJ OY/),c;, y
`
`Opposition a l'encontre du brevet
`EP2 349 203 Bl (Anteis) :
`ESSAIS REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`RA1336402
`Version A
`
`Page 4 sur 6
`
`Les resultats ci-dessous illustrent !'influence de la lidoca"ine en presence de rnannitol et de SOS sur
`!'alteration lors de la sterilisation a la chaleur des proprietes rheologiques de gels d'acide
`hyaluronique de masse rnoleculaire rnoyenne en poids de 3 MDa avec un taux de reticulation X=0,12
`a une concentration de 20 rng/g.
`En presence de sos les effets du mannitol sont detruits par la lidoca'ine a 0,3%.
`Caracteristiques rheologiques mesurees a 25"C
`Avant sterilisation
`Apres sterilisation
`G' (lHz) G" (lHz) Tan 6
`G' {lHz) G" (lHz) Tan 6
`Sucrose 256
`31.55
`0.1233
`187
`25.88
`0.1389
`
`Variation
`G' (lHz)
`-27%
`
`31.31
`
`0.1234
`
`197
`
`26.6
`
`0.1350
`
`-22%
`
`27,3
`
`0,1233
`
`140
`
`21,1
`
`0,1511
`
`-37%
`
`Tableau IV
`
`Sterilisation
`121 ·c-10 min
`
`0,1%
`octasulfate
`3,5% Mannitol
`0,3% Lidoca'ine
`0,1%
`Sucrose 254
`octasulfate
`3,5% Mannitol
`8,5 g/1 NaCl
`
`221
`
`Les resultats ci-dessous illustrent !'influence de la lidoca'ine en presence de mannitol et de MAP a
`une concentration de 0,3 mg/g, sur l'alteratlon lors de la sterilisation a la chaleur des proprietes
`rheologiques de gels d'acide hyaluronique de masse moleculaire moyenne en poids de 3 MDa avec
`un taux de reticulation X=0,12 a une concentration de 20 mg/g.
`En presence de MAP, les effets du mannltol sont detruits par la lidoca'ine a 0,3%.
`Caracteristiques rheologiques mesurees a 25"C
`Avant sterilisation
`Apres sterilisation
`G' {lHz) G" (lHz) Tan 6
`G' (lHz) G" (lHz) Tan 6
`Ascorbyl 264
`38.39
`0.1454
`187
`31.93
`0.1708
`de
`
`Variation
`G' (lHz)
`-30%
`
`Sterilisation
`121 ·c-10 min
`
`0,03%
`phosphate
`magnesium
`3,5% Mannitol
`0,3% Lidoca'ine
`0,03%
`Ascorbyl 270
`phosphate
`de
`magnesium
`3,5% Mannitol
`8,5 g/1 NaCt
`
`221
`
`38.6
`
`0.1431
`
`209
`
`33.63
`
`0.1607
`
`-23%
`
`27,3
`
`0,1233
`
`140
`
`21,1
`
`Tableau V
`
`0,151 1
`. ~7%--~
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`

`vJ V. .I.Yl~.~.Y
`
`Opposition a l'encontre du brevet
`EP2 349 203 B1 (Anteis) :
`ESSA/S REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`RA1336402
`Version A
`
`Page 5 sur 6
`
`Sterilisation
`121 °C - 10 min
`
`Tan o
`
`0.1464
`
`Variation
`G' (lHz)
`
`Tan o
`
`0 .1705
`
`- 28%
`
`Les resultats ci-dessous illustrent !'Influence de la lidoca"ine en presence de mannitol et de MAP a
`une concentration de 0, 7 mg/g, sur !'alteration lors de la sterilisation a la chaleur des proprietes
`rheologiques de gels d'acide hyaluronlque de masse moleculalre moyenne en poids de 3 MDa avec
`un taux de reticulation X=0,12 a une concentration de 20 mg/g.
`En presence de MAP, les effets du mannltol sont detruits par la lidoca"ine 0,3%.
`Caracteristiaues rheolooiaues mesurees a 25°C
`Avant sterilisation
`Apres sterilisation
`G'
`G"
`G'
`G"
`(1Hz)
`(lHz)
`(lHz)
`(lHz)
`0,07% Ascorbyl 264
`190
`32 .35
`41.00
`phosphate
`de
`magnesium
`3,5% Mannitol
`O 3% Lidoca'ine
`0,07% Ascorbyl 266
`phosphate
`de
`magnesium
`3 5% Mannltol
`8,5 g/1 NaCl
`
`38.03
`
`0.1430
`
`208
`
`33.90
`
`0.1628
`
`·22%
`
`221
`
`27,3
`
`0,1233
`
`140
`
`21,1
`
`0,1511
`
`-37%
`
`Tableau VI
`
`Les resultats ci-dessous illustrent !'influence de la lidoca'ine en presence de mannitol sur !'alteration
`lors de la sterilisation a la chaleur des proprietes rheologiques d'un gel d'aclde hyaluronique de
`masse moleculaire moyenne en poids de 3 MDa a une concentration de 20 mg/g, non reticule.
`
`Quelles que soient les concentrations en lidoca"ine, les effets deleteres de celle-ci sur la viscosite des
`compositions conduisent a obtenlr systematiquement des compostions ayant des proprietes plus
`degradees que des compositions sans lldoca'ine.
`Caracteristiques rheologiques mesurees a 25°C
`Avant sterilisation
`Apres sterilisation
`11 (Pa.s)
`·,, (Pa.s)
`1708
`20
`
`Sterilisation
`121 •c - 10 min
`
`3,5% Mannitol
`1,0% Lidoca'ine
`3,5% Mannitol
`0,6% Lidoca'ine
`3,5% Mannitol
`0,3% Lidoca"ine
`3,5% Mannitol
`0,1% Udoca'ine
`3,5% Mannitol
`
`8,5 g/1 NaCl
`
`1670
`
`1696
`
`1660
`
`1673
`
`1407
`
`27
`
`59
`
`215
`
`700
`
`479
`
`Tableau VII
`
`Variation
`Tf
`-99%
`
`-98%
`
`-96%
`
`-87%
`
`-58%
`
`-66%
`
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`

`

`\ly V.tY.f>.~.Y
`
`4) Conclusions
`
`Opposition a l'encontre du brevet
`EP2 349 203 B1 (Anteis) :
`ESSA/S REALISES DANS LE CADRE D'UN AUTRE
`PROJET
`
`RA1336402
`Version A
`
`Page 6 sur6
`
`La presence de lldoca"ine (a partir de 0,3% pour les gels reticules et a partir de 0,1% pour les gels non
`reticules) detruit Jes effets du mannitol sur les proprietes rheologiques du gel a pres sterilisation.
`Aucune formulation comprenant de la lidoca'ine a 0,3 % ne presente une moindre degradation des
`proprietes rheologiques en comparaison a des compositions ne comprenant que le polyol.
`
`L'addition de composes de type derive de vita mine C au derive de sucrose ne modifie pas l'effet de la
`lidoca"inesur les proprietes rheologiques d' un gel apres sterilisation.
`
`5) Validations
`
`Redige par:
`5or· J OCfEHP6
`
`Norn :
`
`Le:
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`Ve rifie par :
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`RODC004 version A
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`©Reproduction et diffusion interdites sans autorisation de VIVACY®
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`