Department of drug Sciences University of Pavia

COMPARATIVE STUDY OF MUCOADHESIVE PROPERTIES OF PRODUCTS FOR ORAL APPLICATION

Contract number: S/ 16/ 12-1-0

Product tested: Aftamed gel cod 1510200 - 2016/09 Emoform Aftagel cod 33942A - 2017 / 12 Wetafte Gel cod 011300- 2018/11 Gum Afta Clear cod 1985 - 2017 / 07 Buccagel Afte cod T23 - 2018/07 Alovex Gel - cod 262541 - 2018/ 09 Anaftin cod 20 '1541 - 2018/07

Client: Cosmetica l talia Servizi s.r.L

Sponsor:

Via Accademia 33 20131 Milano BIO PLAX ITALIA s.r.L Via F.gadi, 7 20144 ~:Wano

Aim of the study

Test supervisionato da COSMETICA ITALIA SERVIZI Via Accademia 33 - 20131 Milano

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The aim of the study was to evaluate "in vitro" mucoadhesive properties of gel formulations for oral

application with different 9uali-t1uantitative composition.

Experimental protocol !11 vitro mucoadhcsive test consists o f the evaluation of mucoadh esive properties by using the

detachement strength measmemcn t of a mucin tablet from the sample sm facc to be tested. Mucin tablet

was obtained by direct compression of the mucin powder, bovine derived- 11 ty pe (Sigma Aldrich, Italin) .

The powder, appropriately weigthed, was trnn ferred in an aluminum matri.x and subjected to a load of

10 tons; the load was maintained for 40 seconds. A hydraulic press was used. The tablet o f mucin had a

diameter of 18 mm, a thickness of 2 mm and a surface area o f 254 mm2.

T he measurements were performed by using a tcnsimeter model Shim adzu AUTOG lv \ PH AGS-D500,

equipped with a loaJ cell o f ·10 g. T he mea urem cat were performed maiJJ tlitining an anaJy- i · tcmpc1:aturc of about 34°C ± 0.5 °C, in order to simulate the physiological conditions of oral cavity. r\ ccording to

protocol:

Department of Drug Sciences - Admin istrative headquarter - Viale Taramelli 12 - 27100 PAVIA Tel. +39-0382-987358-7362 Fax +39-0382-422975 e-mail : emdip07@unipv.it website: http://chifar. unipv.eu/site/home.l1tml Section of Pharmaceutical Chemistry and Technology - Viale Taramelli 12 -27100 PAVIA Section of Pharmacology - Viale Taramelli 14 - 27100 PAVIA

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✓ 5 g o f the sample were distributed homogeneously into a special housing with a diam eter o f 35

mm cm and a thickness of 5 mm and kept 5 minutes to reach temperature,

✓ th e tablet, allocated to the load cell, was suitably hydrated witl1 a mucin solution at 10% (pH

3.70);

✓ i\ ti m e corresponded to 30 seconds was re9uired so that the ·upport does no t sink further into

the sample, modi fying the sta rting analysis conditions, and necessary to ensure possible

interactions between tab let and gel fo rmulation.

T he "pre-loading" was approximately 18 mN. T he force of mucoadhesion was measured by setting speed

of 10 mm/minute. T he m easurem ent was considered concluded when the po lymer was visibly separa ted

from the m ucin tablet. Each sample was analysed in triplicate and resul ts were repor ted as mean and

standard deviation.

Specifically, the maximwn adhesion fo rce (Fmax) and the work o f adhesion (m.J) with respect to the

contact area were evaluated for each sample.

T he comparison was made by:

G el Carbopol 934 2% (positive control)

Artificial saliva solution (negative control)

Product composition

Aftamed gel cod 1510200 - 2016/09 Composition: hyaluronic acid (sodium salt) with high molecular weight (240 mg/ 100g).

Emoform Aftagel cod 33942A - 2017 / 12 Composition: aqua, PVP, taurine, xanthan gum, zinc gluconate, P EG-40 hydrogenated castor oil, sodium saccharin. flavom.

Wetafte Gel cod 011300 - 2018/11 Composition: hyaluronic acid with low molecular weight.

Gum Afta Clear cod 1985 - 2017 /07 Composition: sorbitol, aqua, propanediolo, propylene glycol, polycarbophil (Noveon r\Al USP), Mucosave FG (maltodextrin, Opuntia fi cus-indica Seed Oil, Olea Europeae f ,eaf Lxtract), sodium hydroxide, tauri.ne, PVP Q~uviskol K90), preservative Geogard Ultra (gluconolactone, sodium benzoatc, calcium gluconate), xi.lirol, sodium hyaluro nate, symrclief (bisabolol , 7.ingiber root extract), sucralose, flavours, stevia rebaudiana extract, CI91 40, Cl42090.

Buccagel Afte cod T23 - 2018/07 Composition: triesters peroxidizecl of glycerol (92.70%), silicon dioxide, sodium saccarinate, flavours.

Alovex Gel - cod 262541- 2018/09 Composition: aqua, PVP, maltodexrrin, PEG-40 hydrogenated castor oil, xanthan giun, potassium sorbate, sodium bcnzoate, sodium hyaluronate, bcnzalkonium chloride, flavours, propylene glycol, sodium salt EDTr\, sodium saccharin, D ipotassium glycyrrhizate, r\ loe Barbadensis (Aloe) Leaf Juice.

Ao aftin cod 201541- 2018/07 Composition: aqua, PVP, mal todxtrin, PEG-40 hydrogenated castor oil, xanthan gum, potassium sorbate, sodium benzoate, sodium hyaluronate, benzalkonium chloride, flayours, propylene glycol, sodium salt F:DTA, sodium saccharin , dipotassium glycyrrhizatc, aloe barbadensis (Aloe) leaf juice.

Department of Drug Sciences - Administrative headquarter - Viale Taramelli 12 - 27100 PAVIA Tel. +39-0382-987358-7362 Fax +39-0382-422975 e-mail: emdip07@unipv.it website: http://ch ifar. un ipv. eu/site/home. html Section of Pharmaceutical Chemistry and Technology -Viale Taramelli 12 -27100 PAVIA Section of Pharmacology -Viale Taramelli 14 -27100 PAVIA

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Investigators The cxperimentor is Dr. Priscilla Capra, PhD, degree in Pharmacy. The Study Director is Prof. Paola

Perugini, PhD, Pharmacist, Professor of Cosmetic products at the Deparunent o f Drug Sciences at the

University of Pavia. T he study was carried out at the Department of Drug Sciences, University of Pavia,

Via Taramelli 12, 27100 Pavia, Italy.

Publication

The results of the study may be published only after the written consent of the customer.

11 the information concerning the product in study provided by the customer and not previously

published are co nsidered confidential and it shall remain of only client property. T he investigator agrees

to use this informatio n o nly in relation to the study and no t for any other purpose without a written

consent of the customer. The investigator knows that study data will be used by the sponsor for the

development of the study product and therefore they may be disclosed to other investigators or

Government Agencies. ft is also known that there is an obligation on the part of the inves tigator to

provide the full results of the test and all data collected during the study.

Results and discussion

Evaluation of mucoadhesive properties In Figure 1 results obtained by mucoadhesivc test of gel formulations arc reported. Table 1 shows the

means and corresponded standard deviations o f parameters useful for a comparative assessm ent of the

products: wo rk of mucoadhesion (Lm expressed in mN*mm or mJ) , maximum deformation at

detachment D (mm), maximum force for unit area (a) expressed as mN / cm2.

Jn particular, in Figure 1 re ults corresponded to the work of mucoadhcsion are reported. The work of

mucoadhesion represents the work required to separate two surfaces in contact: it represents the work

that must be achieved to counteract adhesive forces . \ s it is possible to observe in the graph, for almost

all samples the work of mucoadhesion is calculated less than 100 m * mm.Only the Aftamcd sample

profile is significant different: it measures a mucoadhesion work o f about 500 nu'\J+mm (± 54), fifth times

higher compared to the otl1er samples.

T he higher value is to be attributed not only to adhesive forces, but also to cohesive ones. More

considerations can not bl'. made because a detaikJ ljUalitativl'. composition of thl'. sample is not reported.

foi nally, Wctaftc and Anafti.n products, although not comparable to A framed values, arc characterized by

a work o f mucoadhcsion higher than 100 mN, oth erwise from the o tl1er samples.

11 Department of Drug Sciences - Administrative headquarter - Viale Taramelli 12 -27100 PAVIA Tel. +39-0382-987358-7362 Fax +39-0382-422975 e-mail : emdip07@unipv.it website: http://ch ifar.unipv.eu/site/home.html Section of Pharmaceutical Chemistry and Technology - Viale Taramelli 12 -27100 PAVIA Section of Pharmacology - Viale Taramell i 14 -27100 PAVIA

700,0

600,0

I E 500,0 E * 400,0 z E E

300,0 I _, 200,0

100,0 I I :,:

0,0 ~·· ,{v ~" "'~ ;{v+ ~" ~ X

o" t.y'?' ~ "?-(j ~ ..,..0 ~<,; o" c> 0 (.,c.; ':vo ':<."'- ':<."'-"?' ~~ ~~ ':<."'-"" "' "?' 0~ ~ <o-0 '?' '?' c.,O

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Figure 1 - \v'ork o f mucoadhesion (mN*mm). measured for each sample. ·111e analyses were made in triplicate.

In conclu~ion, in Table 1 the significant parameter of the mucoadhesion test for each sample arc

reported.

Table 1 - Mean values and standard deviations of results.

SAMPLE

:\ Fr.-\ CLE:\R Ui\f \VET .\FTE BUCC.-\GEL .AN. FT1N .\LO\'EX -\.FT ·\ GET . :\FT:\ MED CONTROL+ CONTROL -

*Deformation at detachment II J

Reference

Lm (mm'1'mN or mj)

1 13.7 ± 2.9 200.7 ± 73 '1 06.2 ± 16.2 269.6 ± 44.5 89.9 ± 13.5 8 1.S ± :u 532.6 ± 53.9 65.8 ± 6.0 3A ± 0A

o (mN/cm2) D* (mm)

15.0 ± 0.4 55 ± 0.4 10.3 ± 0. 28.8 ± 9.6 17.9 ± 1.0 4.8 ± 0.2 17.9 ± 0.7 15.5±05 11J.4 ± 0.3 6.7 ± 1.3 R.9 ± 0.5 7.7 ± 0.8 10.7 ± 2.2 28.2 ± 0.3 12.6 ± 1.6 4.9 ± 1.2 0.6 ± 0. l 3 .8 ± 0A

f11 I f. I lagers trom and K. Edsman. Interpretation of mucoadhesive properties o f polymer gel preparations using

a tensile strenght 111.c thod. Journal of Pharmacy an f Pharmacology, 2UlH, 53: 1589- 1599.

Department of Drug Sciences -Administrative headquarter- Viale Taramelli 12 -27100 PAVIA Tel. +39-0382-987358-7362 Fax +39-0382-422975 e-mail : emdip07@unipv.it website: http://chifar.unipv.eu/site/home. html Section of Pharmaceutical Chemistry and Technology -Viale Taramelli 12 -27100 PAVIA Section of Pharmacology - Via le Taramelli 14 -27100 PAVIA

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Conclusions

The study had the aim to assess "in vitro" mucoadhesive properties of different gel formulations for oral application.

From results obtained by applying the protocol above described, it can be concluded that:

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The sampies behave differently, in accordance with their qualitative composition.

The Anaftin and t\ftamed gel formulations are characterized by a work of mucoadhesion of 270 and 533 mm*mN, respectively, demonstrating significant mucoadhesive properties. This behavior was also confirmed by the strain values at detachment (mm).

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Date 16th June 2016 Prof. Paola Perugini, Study Director

Department of Drug Sciences - Administrative headquarter - Viale Taramelli 12 -27100 PAVIA Tel. +39-0382-987358-7362 Fax +39-0382-422975 e-mail: emdip07@unipv.it website: http://chifar.unipv.eu/site/home.html Section of Pharmaceutical Chemistry and Technology - Viale Taramelli 12 -27100 PAVIA Section of Pharmacology -Viale Taramelli 14 - 27100 PAVIA