Safety Assessment of Soy Proteins and Peptides as Used in Cosmetics

Transcription

1 Safety Assessment of Soy Proteins and Peptides as Used in Cosmetics Status: Draft Final Report for Panel Review Release Date: August 28, 2015 Panel Meeting Date: September 21-22, 2015 The 2015 Cosmetic Ingredient Review Expert Panel members are: Chair, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G. Marks, Jr., M.D., Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is Lillian J. Gill, D.P.A. This report was prepared by Christina Burnett, Senior Scientific Analyst/Writer, and Bart Heldreth, Ph.D., Chemist CIR. Cosmetic Ingredient Review 1620 L Street, NW, Suite 1200 Washington, DC ph fax cirinfo@cir-safety.org

2 Commitment & Credibility since 1976 Memorandum To: CIR Expert Panel Members and Liaisons From: Christina Burnett, Senior Scientific Writer/Analyst Date: August 28, 2015 Subject: Draft Final Safety Assessment of Soy Proteins and Peptides Enclosed is the draft Final Report of the Safety Assessment of Soy Proteins and Peptides as Used in Cosmetics. (It is identified as soypep092015rep in the pdf document.) At the June 2015 meeting, the Panel issued a tentative safety assessment on soy proteins and peptides with the conclusion that the 6 ingredients listed in the report are safe in cosmetics in the present practices of use and concentration. Since June, no new data have been received. The Council had no comments on the tentative report. The Panel should carefully review the abstract, discussion, and conclusion of this report and issue a Final Safety Assessment L Street NW, Suite 1200, Washington, DC (Main) (Fax) ( ) cirinfo@cir-safety.org (Website)

3 SAFETY ASSESSMENT FLOW CHART INGREDIENT/FAMILY Soy Peptides MEETING Sept 2015 Public Comment CIR Expert Panel Report Status Priority List INGREDIENT SLR Mar 2015 PRIORITY LIST Some of the ingredients in this report were originally part of the general hydrolyzed protein report that was announced May However, the Panel reorganized that report to group the hydrolyzed proteins by source. 60 day public comment period Draft Report DRAFT REPORT June 2015 Table Table IDA TR IDA Notice IDA 60 day public comment period Draft TR DRAFT TENTATIVE REPORT Table Table Tentative Report June 26, 2015 Issue TR 60 day Public comment period Draft FR DRAFT FINAL REPORT Sept 2015 Table Table Different Conclusion PUBLISH Final Report Issue FR

4 Soy Peptides History May 2012 Scientific Literature Reviews announced for Hydrolyzed Source Proteins. December The CIR Expert Panel combined the report with one on source amino acids and called it plant- and animal-derived amino acids and hydrolyzed proteins. The Panel requested additional data to support the safety of 75 plant- and animal-derived amino acids and hydrolyzed proteins. The additional data needed are: (1) method of manufacturing data for both plant and animal-derived amino acids and hydrolyzed proteins, especially for hydrolyzed wheat protein; and (2) composition and characterization specifications of plant and animal-derived amino acids and hydrolyzed proteins, including molecular structure and molecular weight ranges from several suppliers to determine if there is a consistency in cosmetic grade plant and animalderived hydrolyzed proteins, especially hydrolyzed wheat protein. March 2013 and Post Meeting The Expert Panel tabled further discussion on animaland plant-derived hydrolyzed proteins to allow CIR staff to reorganize the report. The staff decided to group the hydrolyzed proteins by source for separate evaluations by the Panel. Hydrolyzed wheat protein and hydrolyzed wheat gluten was the first report reviewed. The review of the other animal- and plant-derived hydrolyzed proteins will be performed sometime in the future. March 2015 Scientific Literature Review announced on soy peptides, including hydrolyzed soy protein and hydrolyzed soymilk protein. June The Panel issued a tentative safety assessment with the conclusion that the 6 soy-based ingredients are safe in cosmetics in the present practices of use and concentration. The Panel noted that soy proteins are known food allergens that can elicit Type I immediate hypersensitivity reactions when ingested by sensitized individuals. However, the Panel was not concerned that such reactions would be induced by dermal exposure, because these ingredients are water soluble, would not penetrate the skin, and have molecular weights that are well below that which would cause IgE-binding. The Panel determined that studies showing little-to-no irritation in ocular animal studies, no dermal irritation or sensitization in animals and human subjects, and no reported cases of Type I immediate hypersensitivity reactions from cosmetic use to support their conclusion for these ingredients.

5 Soy Peptides Data Profile September 2015 Writer, Christina Burnett In-Use Physical/Chemical Properties Method of Manufacturing Composition Genotoxicity Irritation/Sensitization - Nonhuman Irritation/Sensitization - Clinical Ocular/Mucosal Phototoxicity Case Studies Glycine Max (Soybean) Polypeptide X Glycine Soja (Soybean) Peptide X Glycine Soja (Soybean) Protein X X Hydrolyzed Soy Protein X X X X X X X X Hydrolyzed Soy Protein Extract Hydrolyzed Soymilk Protein X X X X X Soy - Generic X X indicates that data were available in the category for that ingredient.

6 SEARCH STRATEGY FOR SOY PEPTIDES (Performed by Christina Burnett) February 2015: SCIFINDER search for soy, soy peptides, or soy protein yielded 0hits. -search for soy proteins peptides polypeptides oligopeptides yielded 7815 hits. - Limit to adverse effect, including toxicity yielded 291 hits. - Further limit to proteins and allergens yielded 76 hits. Many of the references were dietary focused reports. February 2015: PubMed search for soy protein yielded 200 hits, 6 ordered and 2 downloaded. -search for soy protein dermal yielded 1 hit. -search for soy protein skin yielded 5 hits, 1 downloaded. -search for hydrolyzed soy yielded 1 hit. 6 references were ordered. Search updated July No new relevant references identified.

7 Soy Proteins and Peptides June 15-16, 2015 Belsito s Team DR. BELSITO:. Okay, so soy peptides? Okay. So, this is the first time we're looking at this. Six ingredients, function mainly as skin and hair conditioning agents, personal care products. Hydrolyzed soy has the largest number of uses at 862 and the highest reported maximum concentration at 3.5. And hydrolyzed soy protein and hydrolyzed soy milk protein were included in the draft report of plant and animal-derived amino acids and hydrolyzed proteins. And so this is our first look. DR. SNYDER: So, what about the title? This is more than just peptides. It's actually proteins, right? And before, in a previous report, we used the -- somebody's definition of peptide versus protein with a cutoff of 40 amino acids. If it was greater than 40 amino acids it was a protein. If it was less than 40 amino acids, it was a peptide. DR. BELSITO: Well, we had that concern because of IGE binding -- DR. BERGFELD: What -- MS. BURNETT: It was hydrolyzed wheat protein. DR. BELSITO: Yeah. DR. SNYDER: Which we also then referred to -- in the second paragraph, Introduction, here, which I wasn't really certain what the relevance of that paragraph was without it in the context of that sort of discussion about the size, because I -- so I just thought, this is more than just soy peptides, because (inaudible) soy proteins and peptides. So, we probably should change the title. And then in the report, some of the peptides we have listed are actually larger than some of the things we have designated as proteins, which was even further confusion to -- we had a soy peptide at 500 daltons and a hydrolyzed soy protein at 300 daltons. DR. BELSITO: Yes, which I think, you know, beg the issue as to whether we had any information on absorption, which we really don't. And then I also said since these contain phenylalanine, if they're absorbed, would that be an issue for the PKU patients? DR. LIEBLER: I very much doubt that. DR. BELSITO: Doubt they'd be absorbed or be an issue? DR. LIEBLER: Both. DR. BELSITO: But we don't have the data. DR. LIEBLER: Yes. I don't think we have ever seen data for peptides being very well absorbed. I mean, you know, if you're down to about 300 molecular weight, I guess it would be the lowest for these, right? That's about a dipeptide-tripeptide range. DR. KLAASEN: I mean, from the intestine, there is actually a specific transporter that transports di- and tripeptides. Otherwise, they're not absorbed because they're just too water soluble. You know, so absorption of amino acids for a long time was thought you had to get down to one amino acid before it would be actively transported into the blood. But it has been shown in the last couple of decades that there is another transporter that will transport -- it's called the peptide transporter -- that will transport some di- and tripeptides. But this transporter

8 doesn't exist in the skin, and so the likelihood of any of these peptides, even the smallest di- and tripeptides, aren't going to be absorbed by the skin to any significant amount. DR. BELSITO: So, we need to incorporate that somehow -- DR. KLAASEN: That could be. DR. BELSITO: -- into this. DR. SNYDER: So, going back to that second paragraph in the Introduction about the previous reports and the molecular size related to sensitization issues, why were you -- why is that in there? DR. BELSITO: Well, I guess the next issue then becomes: Could this become an issue like hydrolyzed wheat, because soy is a fairly common food allergen, particularly in atopic children. So -- and I think that even though we haven't seen an epidemic like we saw with hydrolyzed wheat in Japan, it doesn't mean that it couldn't occur. So, I mean, I think that when we are looking at these hydrolyzed proteins, we need to be very cognizant about -- particularly when we're looking at a source that's known to be a common food allergen. DR. SNYDER: But my issue is it's not the same protein, hydrolyzed protein, that's the issue. It's not the same hydrolyzed protein in soy that is in corn. DR. BELSITO: No, but I -- the -- DR. SNYDER: I mean, I understand. I mean, I can understand what you're saying. DR. BELSITO: Right. DR. SNYDER: But I just didn't know what -- these don't really have that much relevance to this report other than we're aware that certain hydrolyzed proteins can be allergens. MS. BURNETT: Well, I -- partially this used to be part of a bigger report that we chopped up. So, I think for giving a history, if someone wanted to know what happened to -- if they saw that report for the 20 default then said, well, what happened, you know, they reviewed one, what happened to the (inaudible). DR. BELSITO: Okay, so then, I -- MS. BURNETT: That was part of the reason why I put that there. DR. BELSITO: Okay. DR. SNYDER: So, I think you need to put it in the context that -- MS. BURNETT: Okay. DR. SNYDER: You know, because it just kind of stood there, because when I first read it, I was concerned that you were insinuating that there's -- DR. BELSITO: Well, it may not belong here in the introduction, but it may belong in the discussion. You know -- MS. BURNETT: That could -- that would -- DR. BELSITO: And also I don't -- do you talk about soy reactions in foods in this report? MS. BURNETT: No, I do not. I think I have noted that it is a known food allergen.

9 DR. BELSITO: Right. You may want to put a little paragraph about that and then -- you know, so there's some context, because in the discussion I think we should be cognizant that there was an issue with wheat and that soy is also a fairly common food allergen, and we have a concern that there could be an issue with soy. And it may be -- at least I personally would feel more comfortable putting that same type of molecular weight restriction on soy as we did on wheat. DR. ANSELL: Yes, I would be interested in seeing what the discussion looks like, but we certainly agree that putting the statement about wheat in the introduction just -- it doesn't follow. It's confusing and would need to be put within context as well. MS. BURNETT: By the way, how do we make a comment to this new system? DR. LIEBLER: Well, you hydrolyze -- 13's way smaller than the hydrolyzed wheats that we (inaudible) earlier. So, I mean, you could say it but it's unnecessary. The product as characterized -- the data provided is that it's -- that these are, what was that figure you have, figure 1 or 2, figure 1, you know, up to about 4800 max. But then you've got two sensitization studies that are negative at high-use concentrations. And the highest use concentration we have on the table is around, what 31/2 percent? And the sensitization studies are, like, 25 percent. DR. SNYDER: Yet hydrolyzed soy protein -- we had data neat and 35 percent, and it's only 300 daltons. So, that cutoff won't really work. I don't think it will be applicable here. DR. BELSITO: Okay. Well, we said less than 3500 daltons, right? DR. SNYDER: Correct. DR. BELSITO: But then if we're down to 300 daltons, aren't we worried about absorption? You're saying no because of the KOW. DR. LIEBLER: Because of that and the lack of the transporter that Curt mentioned. DR. BELSITO: Right, okay. Okay, then, you know, what you can do in the discussion is, you know, you talk about type 1 hypersensitivity on page 11 of the PDF, so that's already been brought in. And then in the discussion, we can simply point out that the molecular sizes are, you know, below, well below that where there were issues with hydrolyzed wheat protein, causing urticarial reactions. DR. LIEBLER: You know, with these proteins being able to interact with targets -- for example, IGE -- it's not just a length. I mean, it's the composition and sequence that confers specificity that enables these reactions to occur. I'm unfamiliar with soy allergies, so I don't know, you know, what about them would be instructive in considering their use in possible sensitization, but the data are negative; the size is such that if there were allergies, they wouldn't be able to proceed by the mechanism that we considered for the wheat, so, you know, what we have is -- I think we can simply say, in the discussion we were aware that soy can be a source of food allergies but there is no data to support -- to suggest there's a concern in terms in sensitization for skin. DR. BELSITO: Okay. Under "Use" on page 10 of the PDF you say in the second paragraph, "hydrolyzed soybean protein." However, that's not a name in the introduction. You have "hydrolyzed soy protein." So, should we be consistent? MS. BURNETT: Yes. DR. BELSITO: There are two times in that paragraph where you say -- well, once you say "hydrolyzed soybean protein." Since everyone is satisfied with the manufacturing methods, the impurities, the soy milk we have irritation but we don't have sensitization and we don't have method of manufacturer impurities and we have a compositional

10 breakdown in terms of protein and is everyone comfortable with the soy milk? Does anyone really know what soy milk is? I know you buy it in a store but I mean where does it come from? DR. LIEBLER: I found myself asking the same question because I don't really know. I assumed it's some kind of aqueous extract, but if you added a little information about what soy milk is to help us make the leap from the bean to the -- DR. KLAASSEN: They are little tiny -- DR. LIEBLER: You know, there's so much wisdom out there on the prairie Kurt it just boggles the mind at times. DR. BELSITO: I mean there's got to be information as to how they make soy milk, right? DR. KLAASSEN: Sure. DR. BELSITO: Just to comment as I started looking at these botanicals -- products and there's supplied as solutions in water and he was mentioning today about the fact that with the (inaudible) stem cell cultures and callous cultures they didn't need to add preservatives. When you look at hydrolyzed soy protein can contain parabens and quaturnium 15 the soy milk can contain DMDM hydantoin. When these ingredients are blended into a cosmetic product is it required on the label that they put what the preservatives are in the ingredient that they were using? Or is it only required that they put the preservatives that they've added? DR. ANSELL: The question is if you use an ingredient which has a preservative as an ingredient do you then have to disclose the preservative, no. Unless it's being used as a preservative, no. DR. BELSITO: It's being used as a preservative then what was supplied -- DR. ANSELL: Only in -- if it's used as a preservative within the product itself otherwise it's an incidental additive and nonfunctional. DR. BELSITO: I guess my point is as a clinician when I'm seeing someone who's allergic to quaternium 15 or any preservative I essentially should restrict all botanical products because I don't know what's in that botanical product that's been added to the ingredient. DR. ANSELL: I can't answer the clinical part. DR. SNYDER: So they are only required to designate the preservatives in the final formulation, not in the constituency -- compliance as the final formulation. DR. ANSELL: If it's nonfunctional. DR. BELSITO: It is functional but I guess -- DR. ANSELL: That's why they put it in. DR. GILL: They add it to preserve the final product. DR. ANSELL: Right. DR. GILL: But if it's a preservative for the component, the component use as an ingredient, they don't have to list it. DR. ANSELL: That's my understanding. DR. GILL: So it's in there Don.

11 DR. BELSITO: Theoretically if we were to for instance methylchlorothiazolinone -- preservative where we've actually put limits on the use they could exceed those limits. DR. GILL: Yes. DR. ANSELL: That's a different question. That question relates to safety. The question you asked related to the statutory obligation for ingredient disclosure. One would not in their safety assessment if they had a -- DR. BELSITO: Total concentration of methylchlorothiaaolinone preservative above the restriction they would not be able to use it. DR. ANSELL: They shouldn't use it. DR. GILL: Mm-hm, that's what the panel has advised. That is why the boilerplate language discusses it as it does. DR. LIEBLER: If I'm a manufacturer of an ingredient that I'm going to sell to companies that will make cosmetics and I have an ingredient that's some kind of extract and I add MCI to it, and then the company -- cosmetic company makes a product with that and they don't put in that but they do know that that's present in the ingredient that they bought from me. DR. BELSITO: They don't have to label it. DR. LIEBLER: They don't have to label it? DR. BELSITO: That's what I'm hearing. DR. LIEBLER: Perhaps at best then we could fall back on the guidance that this would be like stacking botanicals. DR. BELSITO: Yeah, what concerns me is in many cases we say "safe as used" for quaternium 15 looking at the concentration that's added to the final product and it might not be safe. I think that we need to start thinking when we are looking particularly at preservatives that are potential sensitizers and could be stacked in there. DR. ANSELL: I think you're mixing the disclosure obligation and the safety obligation. DR. BELSITO: I understand that. DR. ANSELL: If you're purchasing a botanical which contains MCI at concentration high enough that it would exceed the recommended use in the final product one would have to say that that is not conforming with the CIR recommendation. DR. BELSITO: I understand, but with the exception of MI I'm trying to run through my mind all the other preservatives we looked at. We've never set a concentration limit. We've simply said safe as used. MS. FIUME: I actually just looked at quaternium -- it's equal to.2 percent. DR. BELSITO: Okay, for that we have, but what about DMDM hydantoin? DR. ANSELL: Well it could be Dan's point -- is a concern that an ingredient may arise from -- multiple botanicals may have the same ingredient and that needs to be accounted for. I think with the preservative it would be identical. You need to know what your ingredients are. And you need to end up with an acceptable final formulation. DR. BELSITO: I just want to throw that out because I just wanted clarification for myself and I think as we come back and we look at preservatives we need to be aware of that issue at least.

12 DR. LIEBLER: We could perhaps make a slight addition to our botanicals boilerplate because we tend to think of the effect of sensitizers from botanicals and stacking them up in cosmetic products as being essentially what nature put there. If the supplier put it there to preserve their product that they then supplied for us in cosmetic ingredients it's the same issue it's just that the supplier put it there and not nature. DR. BELSITO: I mean just to point out -- I want clarification and now that I've gotten that I think as we look at preservatives we should just be aware of that. MS. FIUME: I'm sorry, Dr. Belsito, actually I looked at the abstract and it said it's used at one percent exclusion for DMDM again it says safe as used. There's no concentration limit in the conclusion for DMDM hydantoin. DR. BELSITO: I mean I'm just -- this has been an eye opener for me looking at this and I certainly will be cognizant as we go -- moved along and add preservatives and rereview preservatives. Getting back to soy so we're going to try and get some more information on exactly what soy milk is or soy milk protein. Probably doesn't differ much from soy protein and then are we going safe as used with this group of ingredients with a discussion that amino acids wouldn't be absorbed through the skin. DR. LIEBLER: Right, that's where I am. DR. KATZ: I have a question before you go safe as used. And this is regarding ocular toxicity. Particularly since some of these products are being used as mascaras or potentially in mascaras. Did you feel you had enough information for that? Because the concentration seems to be higher than what the information was for the tests that were done for ocular toxicity. DR. LIEBLER: Table 7. DR. BELSITO: Ocular irritant on nonhuman studies when tested neat and up to 35 percent for the hydrolyzed soy protein. DR. LIEBLER: About half the studies were in the epi-ocular het cam systems at 20/25/30 percent. DR. BELSITO: And you have hydrolyzed soy protein in and it says just need an ocular irritation study performed under it, it doesn't say what type of study. That's in vitro right? Ocular irritation studies. Non- human. MS. BURNETT: Maximum use concentration eye area products is 3.5 percent. DR. LIEBLER: You have a neat in rabbits. You have two in rabbits actually. One is neat -- one is 25 percent. DR. BELSITO: One is nonirritating. DR. LIEBLER: They are pretty good, they are pretty good, I think the data is strong there. Ocular irritant. There's no ocular toxicity here. DR. BELSITO: Safe as used. No caveats. Discuss the lack of absorption. DR. SNYDER: A little bit of expanded description of the story right. DR. BELSITO: Yeah, soy milk and the fact that the molecular size even though that the soy is a known food allergen -- the molecular size is below that would we be concerned about IGE binding and the types of issues that we would see with hydrolyzed wheat. DR. SNYDER: I think it's more simple than that. There is no data there to suggest that there are any issues.

13 DR. BELSITO: Yeah, there was no data with hydrolyzed wheat until there was an epidemic so I think it doesn't hurt. DR. SNYDER: But I mean between the three data sets we have there's not contact allergens in the compositions we saw right? DR. LIEBLER: We have test data with negative results. DR. SNYDER: Yeah, and we have test data. DR. LIEBLER: That's the point yeah. DR. SNYDER: I think that's -- my opinion is that's an unnecessary level of discussion for this, but it's not a hard and fast thing, but -- DR. BELSITO: I mean I would like to see it pointed out in the discussion that we at least considered it. DR. LIEBLER: Okay, yeah, we can. We considered it and we didn't have to make a supposition based on the molecular weights of the peptides. We have test data with negative results. DR. SNYDER: You didn't respond to my question about the titles. Are we going to leave it as -- DR. BELSITO: I don't know what are you suggesting, Paul? MS. BURNETT: I can confer with Bart if you have suggestions. Do you want to say protein and hydrolyzed -- or proteins and peptides? DR. SNYDER: I just thought -- it says peptides but there is more protein data and more protein than there are peptides. I thought it was a mis -- DR. LIEBLER: I would agree with Paul. I think we could basically say soy proteins. Because hydrolyzed proteins falls within that very reasonably and peptides gives the impression of more specific synthetic small -- a much more chemically defined -- DR. BELSITO: Safety of soy proteins? DR. LIEBLER: Yeah. DR. KLAASSEN: You wouldn't want to have proteins/peptides? DR. BELSITO: We could slash it. Whatever your pleasure is. DR. KLAASSEN: I don't think so. DR. SNYDER: Again I go back to -- the confounding thing to me was their were soy proteins listed that were smaller than the soy peptides that were listed, so that's what kind of precipitated -- DR. KLAASEN: When you have hydrolyzed soy proteins what you really have is soy peptides. DR. LIEBLER: Strictly speaking I agree you are correct Curtis, but peptides can phase the sense of more -- at least to me -- maybe that's because I spend too much time dealing with synthetic peptides. But, it conveys a sense of a much more chemically limited and defined group of products. DR. KLAASEN: That's why I was suggesting.

14 DR. BELSITO: Safety and assessment soy peptides and proteins as used in cosmetics. Would that make everyone happy? DR. KLAASEN: That would make it protein/peptides. DR. BELSITO: Okay. DR. LIEBLER: I don't care. We don't slash our titles. DR. BELSITO: Soy proteins and peptides or peptides and proteins. DR. LIEBLER: I voted for proteins. DR. BELSITO: We can have this discussion tomorrow. Change the title to sorry proteins and peptides or just -- Dan votes for proteins or peptides -- I forget what you voted for. DR. ANSELL: Proteins. I'm abstaining. That's right. My name is A-N-S-E-L-L. DR. KLAASEN: Speaking of abstaining maybe I should abstain because I own a very small farm and we grow soybeans. Maybe there's a conflict of interest since about 1/10th or 1/1,000,000 of the soybeans are ending up as cosmetics. DR. LIEBLER: Any itching, Kurt? DR. KLAASEN: No. DR. LIEBLER: They sold him one. Marks Team DR. MARKS: Shall we move on to soy protein -- Oh, soy peptides. I'm sorry. Yes. The first time we've seen these six ingredients, so Tom, Ron, Ron, the six ingredients okay? DR. SHANK: Okay. DR. MARKS: And what do we have as needs? What can we proceed with at 10 in the report? SPEAKER: I trying to find it wrapped up in this botanicals thing. DR. MARKS: And then we've got some more information Wave 2 besides what's in the May 22nd memo from Christina. DR. MARKS: So, Tom, you said tentative, safe. DR. SHANK: I agree. DR. HILL: How are you coming to that conclusion? Basically everything that soy protein -- there's no data. DR. SLAGA: Would you have the -- all of the preview review that -- amino acids, protein and then on top of that in here you have method of manufacturing, composition and impurities, and to me -- DR. HILL: I've only got method of manufacturing on two out of the -- they don't include genotoxicity.

15 DR. EISENMANN: Did you see Wave 2 soy protein? DR. HILL: I'm pretty sure I did. Let me just see. DR. MARKS: Glycine soy protein. DR. HILL: I thought it was duplicative. MS. BURNETT: Sometime the protein itself you have to hydrolyze? DR. HILL: Yes. I mean it -- if we pick up anything other than soy protein -- DR. MARKS: So for you Ron and Tom? DR. SLAGA: I thought we had sufficient from previous things that we had reviewed that relate to this, that works, and the data and read across through this, and I think we have sufficient. DR. MARKS: Yes. You'd want to capture that, Christina, obviously, in the discussion. Are there -- you are referring to the previous reports on amino acids? DR. HILL: How do you extend amino acids to peptides, that doesn't extend at all? As soon as you've got a dipeptide you no longer need to -- DR. SLAGA: Right. We have -- we reviewed a variable on proteins that were hydrolyzed, and had some peptides and deemed them pretty safe. These are used to -- a lot of these are used in food products. DR. HILL: I don't know what is and isn't used in food products because we don't have that information. And the reason I'm asking is because people do sensitize to soy, or soy allergies. So the circumstance could be entirely analogous to what we saw with wheat. DR. BERGFELD: We have something in the rack behind you, do we? DR. HILL: Yes, we do. DR. BERGFELD: I can see (inaudible) -- DR. HILL: But we lack method of manufacturing for everything except hydrolyzed soy protein, and glycine soja. DR. EISENMANN: Well, the one soy protein used have the molecular weight, 80 percent is 5,000 or less. DR. HILL: Yes. That's the hydrolyzed soy protein -- DR. EISENMANN: No. That's soy protein in the -- MS. BURNETT: Wave 2 DR. EISENMANN: Wave 2, just glycine soja, soybean protein under that trade name, and HPLC more than 80 percent of the protein is less than 5,000. DR. HILL: And so I thought -- DR. EISENMANN: The concentration is greater --

16 DR. HILL: Well no -- Okay, but only 80 percent is less than 5,000. So, that means 20 percent is above 5,000, so that -- DR. EISENMANN: But the material is used -- they sell it as a trade name mixture that contains 0.4 percent of this protein. DR. MARKS: I also, Ron, highlighted the type one, allergic reactions, and there were no reports. So, for me that was reassuring, that there's no reports of any reaction. Again, if we had had the wheat protein and had no reports of reactions we probably would have not even considered it. So I think there has to be, even though there can be a soy allergy, otherwise if there's no report on cosmetic, I would have though by now it would have surfaced, particularly when we were talking about 862 uses of the hydrolyzed soy protein, and 313 uses of the glycine. So for me I can read across and say the rest were safe. And of course the usual irritation sensitization studies we have wouldn't detect immediate reaction anyway. So I really rely on what's the experience of reports. DR. BERGFELD: We'd have to use that in a discussion. DR. MARKS: Yes. That's fine. Because you mentioned, Christina, in here, the wheat protein and the molecular weight of being less than or equal to 35 under Dalton, so if that, Ron, would make you feel more comfortable, we could put that in the discussion as to why we arrived at the safe conclusion. DR. HILL: Yes. DR. MARKS: And then obviously, tomorrow when we ask -- when Wilma ask for a discussion we can bring up that issue, and we can see what the experience of Belsito team, but -- DR. HILL: Is this our motion or theirs? DR. MARKS: It really doesn't matter, we can -- DR. HILL: I know, but I'm asking. DR. SLAGA: It's ours. DR. MARKS: It's ours. DR. HILL: Okay. DR. MARKS: And I can move, and then if you want me to say, in the -- well, it will come up in the discussion that Wilma asked for, and then when we come to the vote, you can vote. DR. HILL: Okay. Fine. DR. MARKS: So, you at least I'll move tomorrow that we move -- we issue a tentative report with a safe conclusion, and no restrictions. DR. BERGFELD: And the idea of moving into this end ones, to move the data for Wave 2? DR. MARKS: Oh. Yes. Absolutely! I can. DR. HILL: I did have a question about Table 1, and what I -- my question I wrote here was, if it's a simple name from soja to max, why did the definitions appear to differ? MS. BURNETT: I'm sorry?

17 DR. HILL: Okay. Let me just find Table 1. If you look at glycine max, we are on page 15. MS. BURNETT: Right. DR. HILL: And then right below it, glycine soja. Okay. The definitions are different but I guess one is probably polypeptide and one is peptide. MS. BURNETT: That I tied to peptide -- DR. HILL: Pardon? MS. BURNETT: One is poly, one is specifically you di- and tri-. DR. HILL: Okay. That's what -- DR. MARKS: Any more comments? DR. HILL: That everything is going to go to max, away from soja. What do you think? DR. EISENMANN: My guess is they are going to just put max in the definition, is the first step. DR. HILL: Okay. Yes. DR. EISENMANN: Rather than change the name. MS. BURNETT: Are we going to get a finished -- like an official monograph change? DR. EISENMANN: I don't know when that's going to happen, so I really don't know. MS. BURNETT: Okay. Because I know this, the whole name came from me, now I'm noticing the two different names, and I asked Bart, and then Bart asked Joanne. And we never got -- officially got -- DR. EISENMANN: It has been my preference, I mean, I've been trying to get them to change, and not put in duplicate names for years, on that. DR. HILL: Okay. And I have another general comment, a sort of question, philosophical question. Let me say -- I've got to find the right one. Let me try -- I'm still getting used to the computer. Yes. It's right before it says, note ingredients with the name glycine soja. The primary focus of the safety assessment of these soy peptide ingredients as used in cosmetics is on the potential for irritation and sensitization from dermal exposure. When we say dermal, do we mean automatically, or do we mean automatically, mucosal membranes, nasal, ocular, lips, genital? Is that always automatically implied, is that what we are saying? MS. BURNETT: I believe so. DR. HILL: Because we have -- yes, we have -- I don't know if this is all the ingredient, but I'm seeing now, we have some sort of boilerplate language, where it's, okay, we eat this things, and so we all have a potential that's dermal exposure, and so our only word is irritation and sensitization, which I will again say, eating is not the same thing as exposing the skin because of our digestive tract and first pass metabolism, but that's my question is. When we say dermal exposure are we always including all potential risks? DR. MARKS: Read that sentence again? DR. HILL: It's that, primary focus -- Well, let's go back a sentence.

18 DR. MARKS: Well, that's okay. Primary focus, I mean, you're questioning dermal, we could in there, we could substitute in the uses as documented in this report. Then that covers it. DR. HILL: Yes. Because this seems to be now, some sort of a boilerplate, and maybe I let this go as a boilerplate, but I've just -- DR. MARKS: No. I hear you. I mean we talked about the -- DR. EISENMANN: So, would you be more comfortable saying the focus is not systemic toxicity. I mean, some way -- in some way to -- well -- DR. HILL: You can get systemic toxicity from non- oil exposure and, again, in some cases our digestive tract take things out that wouldn't be taken out when we apply them to the skin. I'm not worried about that with this set of ingredients, but we just seem to be -- we seem to have this boilerplate where, okay, we eat the stuff and so anything that we do on the skin, it's not likely to provide exposure, and therefore we only have to worry about irritation and sensitization, which I think is crap. So, what -- and so I don't like the implication of that in general, it may be true for any given category of ingredients, but we are writing off inhalation exposure, we are writing mucous membrane exposure if we just say dermal. And I'm not comfortable with that implication when it isn't true, in some cases. DR. MARKS: If we delete that sentence would it change the meaning significantly, because then once we delete it, you know, we are focused on all those other things you mention. I would say we consider all the others, so that if we delete the sentence then does that resolve it? MS. BURNETT: The previous sentence says, thus the systemic toxicity potential of soy peptides, via oral exposure is not addressed further in this report. DR. HILL: If we'd just take it out and it makes sense, I'd be happy. But we are using it as some sort of a boilerplate now, so that's why I'm bringing it up now, because to my attention. MS. BURNETT: It's also in, I think, the citrus report, and the previous reports I've worked on. DR. MARKS: Ron Shank, now it's up to you to just delete that sentence. DR. DEWAN: Maybe you could just -- it's been there and used in many reports. It's not going to be one. DR. DEWAN: We've put it in a number of other -- that's okay going forward within, but I -- MS. BURNETT: Delete that? DR. MARKS: I mean the focus doesn't mean that we aren't considering the others, it's just we are looking at that with a more -- DR. HILL: I agree with you, but I don't like to read between the lines, the implication is just writing off everything else without even thinking about it, which is the way reads to me. DR. MARKS: Ron Shank, that's what it says? DR. SHANK: I know it's not what it says, but that's what it implies, implications matter. DR. SLAGA: That's reading the lines.

19 DR. HILL: Implications matter. And sometimes they matter a lot. DR. MARKS: So, what I would suggest again, it sounds like Ron Shank and Tom, you are quietly leaving it as stated. DR. SHANK: I would -- DR. MARKS: So why don't, Ron, you bring that up tomorrow and you'll get the input of the other team, and then we can proceed from there. Then obviously, your concern about that wording is documented now in our minutes, and they'll be documented tomorrow. Full Panel Meeting DR. BERGFELD: Moving on to the next ingredient. Dr. Marks, the soy ingredients, peptides. DR. MARKS: So, this is the first time we've seen this draft report on the soy peptides. There were six ingredients. Our team felt that these ingredients were safe, so we move a tentative report with a safe conclusion be issued. DR. BELSITO: Second, second. DR. BERGFELD: Any discussion points or comments? Don? DR. BELSITO: Yes. So, you know, of course with hydrolyzed soy we were concerned about the same issue we were with hydrolyzed wheat, but the data clearly shows that this is the size of these particles, is all below a size that would bind IG receptors, and the data for contact, or the carrier was clear, so we didn't have that concern. DR. BERGFELD: Did you have a concern of -- DR. BELSITO: But should be brought into discussion. DR. BERGFELD: Thank you. Into the discussion it will go. Anything else? DR. BELSITO: Yes. There was a comment by Paul or Dan, I can't remember, that the title should be changed to soy proteins and peptides, since they are not all peptides. DR. LIEBLER: Between Curt and Paul and myself, we had three different titles. They all had proteins or peptides. One had a slash in it. I voted against the slash, I thought soy proteins might be better, simply because peptides connote selective production of very specific sequences, and that's not what these ingredients really are there. They are hydrolysis products or soy proteins, and so that's why I suggest soy proteins, but my colleagues may want to say something. DR. BERGFELD: Paul? Curt? DR. KLAASEN: In your mass effect technology that you are doing, what are you measuring? DR. LIEBLER: We are measuring peptides. But that doesn't have anything to do with this report. (Laughter) Follow the way. DR. KLAASEN: I don't -- I like, better to have proteins and peptides, but I'm not going to fight over it because --

20 DR. LIEBLER: I'm good with proteins and peptides. If it's just peptides, it makes it sounds like there is, you know, a limited range of synthetic or highly-selected sequences, and that's not what I wanted to convey. But, you know, I'll get over it. Okay? DR. BERGFELD: Ron Hill? DR. HILL: Yes. I mean, the concern that I raised yesterday, and I just want to make sure we briefly discuss again, is that from the work on wheat, we got very great information about what it takes to get that kind of sensitization, and the relationship between dietary consumption and sensitization, versus the cosmetic. And I guess the peptides that might be produced, and then enzymatic hydrolysis will not necessarily be the same as the peptides we would produce when we digest soy protein in our gut. So it's well to keep that in mind, and make statements accordingly. I flagged a couple in the document, to make sure that we were clear on that point. DR. BERGFELD: Thank you. Any other comments? I'm going to move the question. All those in favor of safe? Unanimous. Okay. (Motion passed unanimously)

21 Safety Assessment of Soy Proteins and Peptides as Used in Cosmetics Status: Draft Final Report for Panel Review Release Date: August 28, 2015 Panel Meeting Date: September 21-22, 2015 The 2015 Cosmetic Ingredient Review Expert Panel members are: Chair, Wilma F. Bergfeld, M.D., F.A.C.P.; Donald V. Belsito, M.D.; Ronald A. Hill, Ph.D.; Curtis D. Klaassen, Ph.D.; Daniel C. Liebler, Ph.D.; James G. Marks, Jr., M.D., Ronald C. Shank, Ph.D.; Thomas J. Slaga, Ph.D.; and Paul W. Snyder, D.V.M., Ph.D. The CIR Director is Lillian J. Gill, D.P.A. This report was prepared by Christina Burnett, Senior Scientific Analyst/Writer, and Bart Heldreth, Ph.D., Chemist CIR. Cosmetic Ingredient Review 1620 L Street, NW, Suite 1200 Washington, DC ph fax cirinfo@cir-safety.org

22 ABSTRACT The Cosmetic Ingredient Review (CIR) Expert Panel (Panel) reviewed the safety of soy proteins and peptides, which function in cosmetics primarily as hair conditioning agents and skin conditioning agents-miscellaneous. The Panel considered relevant data related to these ingredients. The Panel concluded that soy proteins and peptides are safe in cosmetics in the present practices of use and concentration described in this safety assessment. INTRODUCTION Soy protein and peptide ingredients function mainly as skin and hair conditioning agents in personal care products. 1 This report assesses the safety of the following 6 soy ingredients: Glycine Max (Soybean) Polypeptide Glycine Soja (Soybean) Peptide Glycine Soja (Soybean) Protein Hydrolyzed Soy Protein Hydrolyzed Soy Protein Extract Hydrolyzed Soymilk Protein The safety of several hydrolyzed proteins as used in cosmetics has previously been reviewed by the Panel. The Panel concluded that hydrolyzed collagen and hydrolyzed corn protein are safe for use in cosmetics. 2-6 Additionally, the Panel concluded that hydrolyzed wheat gluten and hydrolyzed wheat protein are safe for use in cosmetics when formulated to restrict peptides to a weight-average MW of 3500 Da or less. 7 Soy proteins and peptides are used as food. The U.S. Food and Drug Administration (FDA) determined that the use of peptones as direct food substances is generally recognized as safe (GRAS) and that soybean protein is GRAS for substances migrating to food from paper and paperboard products. Because daily exposure from food use would result in much larger systemic exposures than from use in cosmetic products, the systemic toxicity potential of soy peptide ingredients via oral exposure is not addressed further in this report. The primary focus of the safety assessment is the review of safety based on topical exposure Note: Ingredients with the name glycine soja (soybean) are undergoing a name change to glycine max (soybean). Both names refer to the same plant species. 8 This change represents the more commonly accepted nomenclature. CHEMISTRY Definition The definitions of the soy peptide ingredients included in this report are provided in Table 1. The soy protein and peptide derivatives form a broad category of materials which are prepared by extraction from soy and partial hydrolysis to yield cosmetic ingredients. Soy proteins and peptides can also be separated on the basis of molecular size. By removing oil at lower temperatures, soy protein isolate is obtained, and is widely used in the food industry Whole aqueous extractable soybean proteins can be separated into storage globulin and whey fractions by acidification. The FDA defines the term protein to mean any α-amino acid polymer with a specific defined sequence that is greater than 40 amino acids in size. 12 The FDA considers a peptide to be any polymer composed of 40 or fewer amino acids; however, these definitions of protein and peptide are not necessarily adhered to in the naming of cosmetic ingredients. The acid-precipitable fraction of whole aqueous extractable soybean proteins includes the major soybean storage proteins. The remaining part consists of the minor globulin, γ-conglycinin, and contaminating proteins, including whey proteins. Whey proteins are composed of lipoxygenase (102 kda), bamylase (61.7 kda), lectin (33 kda), and Kunitz trypsin inhibitors (20 kda). The proportion represented by these whey proteins in the acidprecipitated globulins is unknown. Soy protein isolate is a mixture of various proteins, and the main ingredients are classified into four protein categories according to their sedimentation coefficients 2S, 7S, 11S, and 15S which sediment at different gravitational forces when the solution is subjected to a centrifugal field. Among these proteins, 7S (β-conglycinin) and 11S (glycinin) represent 80%-90% of all soybean protein, and the ratio 7S/11S has been reported to be about depending on varieties. The 7S globulin consists of three subunits α (ca 67 kda), α' (ca 71 kda) and β (ca 50 kda). The 11S globulin is a hexamer, and is made up of five different subunits, each of which consists of an acidic subunit A (ca 35 kda) and a basic subunit B (ca 20 kda), linked by a disulfide bond. The 11S globulin was found to dissociate into 2S, 3S or 7S forms in solutions of various ph values and ionic strengths. Amino acid compositions of β-conglycinin and glycinin have been analyzed, but the three dimensional structures are not well established in spite of many efforts.

23 Chemical and Physical Properties Available information concerning chemical and physical properties, including molecular weights, for soy protein and soy peptide ingredients is presented in Table 2. Soy Peptide Dipeptide- or tripeptide-rich forms of soy peptide were described to have a mean molecular weight of around 500 Da. 13 Hydrolyzed Soy Protein A histogram showing the approximate distribution of molecular weights for a hydrolyzed soy protein product from one supplier is shown in Figure The figure shows that approximately 35% of the molecular weight distribution falls between 490 and 1030 Da. One source has indicated the average molecular weight is 300 Da 15 ; however, other sources have reported the molecular weight of their hydrolyzed soy protein product to be approximately Da Hydrolyzed Soymilk Protein A supplier has reported the molecular weight of their hydrolyzed soymilk protein product to be approximately Da. 18 Method of Manufacturing Glycine Soja (Soybean) Protein Glycine soja (soybean) protein can be prepared from defatted low-heat soybean meal. 10 A dispersion of soy flour is prepared by adding distilled water (1: l5, w/v) with final protein concentration of 3.1% (w/w). Then, 2 mol/l NaOH is used to adjust the dispersion to ph 8.5. The dispersion is stirred for 1 h at room temperature and then centrifuged (10000 g, 20 min). The supernatant is adjusted to ph 4.5 with 2 mol/l HCl and centrifuged (10000 g, 20 min). The obtained sediment is resuspended with distilled water (1: 5, v/v) and adjusted to ph 7.0 with 2 mol/l NaOH. Then it is dialyzed against deionized water and freeze-dried. A supplier has indicated that a glycine soja (soybean) protein product is manufactured by adding water to the seed of Glycine max Merrill (Leguminosae), extracting, and then filtering. 19 The solvent 1,3-butylene glycol is then added to the filtrate. More than 80% of the resulting protein product has a molecular weight less than 5000 Da. Hydrolyzed Soy Protein A manufacturing flow chart from a hydrolyzed soy protein supplier is shown in Figure 2. The preparation of hydrolysates can be afforded via acid and enzyme. 10 The above glycine soja (soybean) protein dispersion (4% w/v) is adjusted to ph 2.0 with 1 mol/l HCl, and incubated at 37 C for 30 min. Then, an enzyme (such as pepsin) is added to each part at an enzyme to substrate ratio of 0.3% (w/w) to start the enzymatic hydrolysis reaction. Each fraction is incubated at 37 º C ( min) and the enzyme is deactivated by adjusting the ph to 7.0 with 2 mol/l NaOH. A supplier has reported that hydrolyzed soy protein is produced from isolated soy proteins that are hydrolyzed with a protease enzyme for 2 hours. 20 The enzyme is inactivated by heat once the target molecular weight is achieved. The resultant solution may then be concentrated. Another supplier reported that hydrolyzed soy protein (MW = 300 Da) may be prepared by both alkaline and enzyme hydrolysis. 15 These processes occur for several hours until the desired molecular weight is reached. The final product is a 25% water solution of hydrolyzed soy protein. In a hydrolyzed soy protein product of 8.5%, the supplier states that enzymatic hydrolysis yields a material with 87% of the proteins with a molecular weight less than 2000 Da, 11% of the proteins with a molecular weight between 2000 and 5000 Da, and 2% of the proteins with a molecular weight greater than 5000 Da. 21 Hydrolyzed Soymilk Protein While method of manufacturing for hydrolyzed soymilk protein was not discovered in the published literature nor was information provided by industry, soymilk is reported to be made through water extraction of whole soybeans. 22

24 Composition and Impurities A supplier has stated that hydrolyzed soy protein and hydrolyzed soymilk protein do not contain fragrance ingredients designated as sensitizing by the European Union s cosmetics regulations Glycine Soja (Soybean) Protein A supplier states that glycine soja (soybean) protein product (>80% has MW less than 5000 Da) is comprised of 0.4% protein, 30% butylene glycol, and 69.6% water. 19 Soy Peptide The amino acid composition of a soy peptide sample is presented in Table 3. The analysis found that soy peptide is rich in aspartic acid (12.6%) and glutamic acid (22.1%). 13 Hydrolyzed Soy Protein A supplier states that hydrolyzed soy protein (MW = 300 Da) has heavy metals, arsenic, and iron at levels < 10 ppm, 1 ppm, and 10 ppm, respectively. 15 Another supplier states that hydrolyzed soy protein (87% has MW < 2000 Da) did not have detectable levels of alkaloids, pesticides, or aflatoxins. 21 No traces of arsenic or mercury were detected. Other metals were reported as cadmium (2 ppb), chromium (57 ppb), cobalt (15 ppb), nickel (0.7 ppm), lead (60 ppb), and iron (< 10 ppm). The compositional breakdown of products containing 25% and 35% hydrolyzed soy protein included 74.6% or 64.6% water, respectively, and 0.2% methylparaben, and 0.2% quaternium-15, each. 23,24 Hydrolyzed Soymilk Protein The compositional breakdown of a product containing 21.5% hydrolyzed soymilk protein included 76.6% water, 1.2% phenoxyethanol, and 0.7% DMDM hydantoin. 25 USE Cosmetic The safety of the cosmetic ingredients included in this safety assessment is evaluated on the basis of the expected use in cosmetics. The Panel utilizes data received from the FDA and the cosmetics industry in determining the expected cosmetic use. The data received from the FDA are those it collects from manufacturers on the use of individual ingredients in cosmetics by cosmetic product category in its Voluntary Cosmetic Registration Program (VCRP), and those from the cosmetic industry are submitted in response to a survey of the maximum reported use concentrations by category conducted by the Personal Care Products Council (Council). According to the 2015 VCRP data, hydrolyzed soy protein has the most reported uses in cosmetic products of the ingredients listed in this safety assessment, with a total of 862; about half of the uses are in non-coloring hair products (Table 4). Glycine soja (soybean) protein has the second greatest number of overall uses reported, with a total of 313; a third of those are used in leave-on skin care products and another third are used in hair dyes and colors. The results of the concentration of use survey conducted in 2014 by the Council indicate hydrolyzed soy protein has the highest reported maximum concentrations of use; the maximum concentration used is 3.5% in mascara. Glycine soja (soybean) protein is used at up to 0.9% in eye lotion. No use concentrations were reported for the remaining 4 ingredients. Based on the VCRP data and the results of the Council s concentration of use survey, glycine soja (soybean) peptide and hydrolyzed soy protein extract are not in use. As mentioned previously, some of these ingredients are used in products that are used near the eye. Additionally, some of these ingredients may be used in products that can be incidentally ingested or come into contact with mucous membranes. For example, hydrolyzed soy protein is used in bath soaps and detergents at up to 1.5%. Additionally, some of these ingredients were reported to be used in hair sprays and body and hand sprays and could possibly be inhaled. For example, glycine soja (soybean) protein was reported to be used in body and hand sprays at a maximum concentration of 0.07%. In practice, 95% to 99% of the droplets/particles released from cosmetic sprays have aerodynamic equivalent diameters >10 µm, with propellant sprays yielding a greater fraction of droplets/particles below 10 µm compared with pump sprays Therefore, most droplets/particles incidentally inhaled from cosmetic sprays would be deposited in the nasopharyngeal and bronchial regions and would not be respirable (i.e., they would not enter the lungs) to any appreciable amount. 28,29 The soy peptide ingredients in this report are not restricted from use in any way under the rules governing cosmetic products in the European Union. 26