What are Copper Peptides?

We have had many questions about various assertions coming from people at Procyte Corporation.

1. Copper peptides are complexes of copper ions and small pieces of proteins called peptides. For skin regeneration, the biologically active copper (+2) ion is most studied.

2. All peptides avidly bind copper (II) with binding affinities of about pK=9. This means that for each billion copper ions only one copper ion would not be bound to a peptide. Also, since copper (II) often binds to two peptides and this further raises the binding affinity by about 5 orders of magnitude, the amount of unbound copper (II) is immeasurably small and of no biological consequence. The knowledge that peptides and copper avidly bind to each other is over 100 years old and known by all educated chemists and biochemists.

3. Forming the complexes is extremely easy. A solution of any common copper salt, such as copper chloride or copper sulfate, is dissolved in water. The copper solution is then mixed with at least equimolar peptides and the pH slowly raised by the addition of a base such as sodium hydroxide. As the pH is raised above 5, the formation of copper complexes is virtually instantaneous. If copper salts existed, the copper (II) would immediately form a white precipitate of copper hydroxide.

4. My background and qualifications for making these statements is at www.skinbiology.com/copperpeptideregeneration.html.

Strange Science at Procyte Corporation

We have had many questions about statements and emails coming out of ProCyte Corporation about copper peptides such as the following:

This includes material such as the following:

"Not All Copper Ingredients Yield the Same Results"

For example, copper salts and random amino acid sequences in a formulation are not true, complexed copper peptides, but just plain copper salts and peptides or proteins mixed together (i.e. hydrolyzed soy protein and copper chloride). It takes a special process and the correct amino acid chain to formulate the copper peptide most closely resembling one of the body's own natural copper carrier systems.

ProCyte's GHK copper and AHK copper, which are protected by ProCyte's patents, are the only true copper peptides on the market. GHK is one of the body's natural carriers of copper. Using advanced pharmaceutical processes, these specific amino acid chains bind the copper and protect it so that it can get to where it needs to go and then be recognized by the body as a natural transporter and release the copper. Copper salts are generally irritating and not easily utilized by the body.

ProCyte has over 15 years of research and development behind the GHK copper peptide complex used in Neova Therapy products. The data supporting the improved wound healing outcomes as well as the dramatic improvement in fine lines and wrinkles are specific to studies using GHK:Cu copper.

Protect your investment by purchasing the following brands, which are the only brands that utilize the GHK:Cu copper: Neova, Complex Cu3, Tricomin, Graftcyte, and Iamin (Comment - It is spelled "Iamin" - get a Greek dictionary)."

1. As stated in the opening section, it is impossible to mix an aqueous mixture of the copper ions and equimolar peptides and not form copper peptides as the pH is raised toward neutrality. If the management at Procyte has found a method to do this, they should have it published in chemical journals. This would overturn 100 years of chemical research on the interactions of copper ions with peptides. The entire scientific world would be amazed.

2. There are no advanced pharmaceutical methods needed to for copper chelates. I prepared the first copper complexes of GHK in 1970 and needed no special processes.

3. Technology advances. Propellor planes have been replaced by jets. GHK will be replaced in time by improved complexes for the same uses.

This email was recently sent to Skin Biology by the recipient

Subject:
Re: Our new order
From:
"Robin Carmichael" <rcarmichael@procyte.com>

To:...............

CC:
"Jack Clifford" <jclifford@procyte.com>

Ok.  I will get the order going and will note the new freight forwarder.

Regarding Skin Biology, please note this is NOT a real copper peptide product. (Comment - This is not true - See above) It is a imitator and is not the patented GHK Copper that uses  the body's own natural carrier for copper.  Dr. Pickart was one of the early founders of ProCyte, but he has not been with the company or involved in the technological development of the GHK Cu or any product for over 10 years!!!!!!! (Comment - I have been working on developing an improved technology for copper peptides for the last 10 years, on copper peptides for 34 years, and on aging reversal methods for 40 years)

I do not know a good analogy for you, but basically, ProCyte makes the GHK copper peptides under pharmaceutical control as these require very specific and controlled processing.  Due to the rapid success of ProCyte's patented copper peptide technology, we are now seeing a number of lame imposters (Comment - It is spelled "impostors", get an English dictionary) using various copper substances or salts starting to creep up in the market place.(Comment - so much time has passed that many key patents at Procyte will begin to expire in 2004).

To briefly summarize the differences between copper products:
It is important to note that not all copper peptide products are the same!  For example, copper salts and random amino acids in a formulation are not true, complexed copper peptides, but just plain copper salts and proteins mixed together - rather like sugar and water - do not necessarily make caramel!!!  It takes a special process and the correct amino acid chain to formulate the copper peptide most closely resembling the body's own natural copper carrier system (Comment - All peptides avidly chelate ionic copper (+2), maybe she should stick to making caramels).

ProCyte's GHK:cu copper and AHK:cu copper which are protected by ProCyte's patents are the only true copper peptides on the market.  GHK is the body's natural carrier of copper.  Using advanced pharmaceutical processes, these amino acid chains are complexed such that they bind the copper and protect it so that it can get to where it needs to go and then be recognized by the body as natural transport and release the copper.  Copper salts are generally irritating and not easily utilized by the body.

True GHK Copper that still has the peptide bound to it as the protective carrier is BLUE!!!!  Copper salts + random amino acid products are green- not blue, so it is easy to tell an imposter (Comment - It is spelled "impostors", get an English dictionary).

So remember - copper is vital to skin health and tissue repair, but you need the proper copper peptide complex - GHK:cu - and formulation to allow the copper to be of value.

Hope this helps!!!!

(Comment - Ms. Carmichael should go back to school and learn some advanced chemistry and biochemistry. Or perhaps she should publish these strange ideas in a chemical journal. This would overturn 100 years of chemical research on copper ions and peptides. All peptides avidly chelate ionic copper (+2))

Taken from a manuscript I wrote:

Background

Certain types of copper peptide complexes possess both tissue protective and repair properties. Most information on these effects is based on a human copper peptide complex, glycyl-l-histidyl-l-lysine:copper (II) or GHK-CU.

Research by Dr. Loren Pickart found, between 1983 and 1989, that GHK-CU had protective and regenerative actions on several organ systems including skin, hair follicles, bone, gastric mucosa and intestinal linings. These types of copper complexes are increasingly used in cosmetic skin and hair care products, and after dermatological skin renewal procedures, such as, chemical peels, laser resurfacing, and dermabrasion, to improve post-treatment skin recovery.

The first generation copper peptides invented (circa 1993 to 1988) by Dr. Pickart (known as “Pickart’s 1st Generation Copper Peptides”) performed well in many controlled tests; however, these products failed in FDA clinical trials on the healing of very difficult-to-heal human wounds. This created the need for improved copper peptides.

The flaws in GHK-CU for commercial use are as follows:

1.1 Fragility of the GHK molecule

GHK is very sensitive to enzymatic breakdown.

GHK was discovered in 1973 by Dr. Loren Pickart and determined that the factor was a tripeptide containing glycine, histidine and lysine. (Pickart, A tripeptide in human plasma that increases the survival of hepatocytes and the growth of hepatoma cells. Ph.D. Thesis in Biochemistry, University of California, San Francisco, 1973;  Pickart and Thaler, Nature New Biol 1973 May 16;243(124):85-7,  Tripeptide in human serum which prolongs survival of normal liver cells and stimulates growth in neoplastic liver. )

At this time, only microgram amounts of the peptide were available for further analysis and isolation was very slow and tedious. In 1974, Pickart arranged with Dr. David Schlesinger at the Chemistry Department of Harvard University to determine the actual sequence of amino acids by controlled enzymatic breakdown of the tripeptide. Schlesinger’s group was considered the best for such types of analysis.

Schlesinger had great difficulties in the structure determination of GHK. The procedure used a timed breakdown of GHK by carboxypeptidase to determine the sequence of amino acids in the tripeptide. However, the first two attempts to determine the structure failed because of the extreme sensitivity of the tripeptide to breakdown by carboxypeptidase. Because of the slow rate of tripeptide isolation, this delayed to structural determination by two years. (Growth modulating serum tripeptide is glycyl-histidyl-lysine. Schlesinger, Pickart, Thaler, Experientia 1977, 33(3):324-5)

1.2. Breakdown of the GHK molecule in serum and wound fluid

Carboxypeptidase and other proteolytic enzymes exist in blood plasma, serum and in wound fluids. Studies of radioactive GHK-CU at the Virginia Mason Research Center in Seattle between 1980 and 1985 found that the molecule was rapidly degraded in murine or human serum at 37 degrees centigrade. Localized injection found that  was cleared from the dermis within about a minute.

This fragility and rapid breakdown of GHK and other simple copper peptide complexes is the major problem in developing such types of products for clinical and cosmetic use. In the human body, the  complex can be generated constantly. However, when used as a single dose therapy, its fragility leads to rapid breakdown, clearance from the dermis, and a loss of effectiveness.

1.3 Chelation to copper does not protect GHK from breakdown

It was considered possible that when GHK was chelated to copper that it would become resistant to breakdown. However, this does not appear to be the case.

In human plasma and wound areas, GHK is likely to exist as a mixture of GHK and GHK-CU. GHK has high binding affinity for copper (II) - pK=16.2 - very close to the affinity for copper (II) of albumin. GHK effectively competes with albumin for copper. However, under physiological conditions only about 5% to 20% of GHK molecules would be expected to exist as GHK-CU complexes with copper (II).

Furthermore, despite GHK’s high binding affinity for copper (II), proton magnetic resonance measurement indicated that GHK-CU continually exchanges copper with other GHK molecules. This means that naked GHK is periodically exposed to enzymes that can degrade the molecule. (Kwa E, Bor-Sheng L, Rose N, Weinstein B, Pickart L., PMR studies of Cu(II) and Zn(II) interaction with glycyl-l-histidyl-l-lysine and related peptides, Peptides 1983, 8, 805-808; Elizabeth Kwa, Ph.D Thesis, Department of Chemistry, Univ. of Washington, 1983)

1.4 Rapid removal from sites of injection

As mentioned above, localized injection found that GHK-CU was cleared from the dermis within about a minute. There is a need for a longer acting skin regenerative copper peptide.

1.5 GHK-CU does not adhere to skin or tissues

GHK-CU does not adhere to skin or tissues. Aqueous solutions of molecule just run off intact skin, wounds, and tissues. If the adherence to tissues could be increased, this would, in all likelihood, enhance the persistence and activity of the molecule.

The reason for this lack of adherence is that GHK-CU is a highly charged molecule. This means that it will avidly form hydrogen bonds to water, thusly creating a “shell-of-hydration” in water. Such types of bonding can be very strong, up to 300 Kilocalories per mole. This shell of water molecules will reduce interaction with skin and tissues.

The GHK-CU molecule is very hygroscopic, that is, it will pick up water from the air and turn into a watery syrup. The determination of the 3-dimensional structure of GHK-CU was complicated by this hygroscopic behavior. (Pickart, Freedman, Loker, Peisach, Perkins, Stenkamp, Weinstein, Nature 288, 1980, 715).

2.0 Design of more stable  synthetic analogs

A variety of chemical modifications to GHK have produced bioactive copper complexes with enhanced breakdown resistance.

Pickart invented a series of analogs of GHK-CU that possessed increased resistance to enyzymatic proteolysis. These analogs included GHK-n-ocytyl ester, GHK-amide, and GHK-benzyl ester. When complexed to copper 2, these analogs had healing activities comparable to GHK-CU. (US Patent 4,665,054, filed Feb. 8, 1985, US Patent 4,877,770, filed Oct. 31, 1989)

Dalpozzi et al prepared a partially retro-inverso analogue of GHK was synthesized, in which the -CONH- bond between histidine and lysine was modified as -NHCO-. The new peptide analogue showed approximately a ten-fold increase in stability versus the parent peptide. (Dalpozzo A, Kanai K, Kereszturi G, Calabrese G., H-Gly-His psi (NHCO)Lys-OH, partially modified retro-inverso analogue of the growth factor glycyl-L-histidyl-L-lysine with enhanced enzymatic stability, Int J Pept Protein Res 1993 Jun;41(6):561-6))

Conato et al synthetically modified GHK with breakdown resistant groups. Histidine was replaced with either a synthetic amino acid, L-spinacine, or L-1,2,3,4-tetrahydro-isoquinoline-3-carboxylic acid. The compounds were investigated with potentiometry, solution calorimetry, UV-VIS spectrophotometry, circular dichroism and electron paramagnetic resonance spectroscopies. All the ligands formed copper complexes having different stoichiometries and stabilities. After 3 hours in serum, GHK was degraded but the synthetic compounds showed no significant degradation. (Conato et al, Copper complexes of glycyl-histidyl-lysine and two of its synthetic analogs: chemical behavior and biological activity, Biochim Biophys Acta 1526, 199-210, 2001).

2.1 Problems with synthetic GHK-CU Analogs

The problem with this classical organic chemistry approach is that each new chemical becomes, in FDA regulatory terms, a new chemical entity. This increases the possibility of undesirable side effects and much slower regulatory approvals. This would require extensive safety testing.

Also, an analysis of the possible copper-binding conformations suggests that some of these complexes may lose GHK-CU’s anti-inflammatory activity.

3.0 Pickart 2nd Generation Copper Peptides

To design improved tissue regenerative copper peptides, Pickart set the following goals that the improved copper peptides should:

3.1 Be resistant to breakdown by enzymes in serums and wound fluids.

3.2. Have a high adherence to skin and tissues.

3.3 Use peptides with long records of safety in human use.

3.4 Not require FDA testing as new products.

3.5. Demonstrate skin regenerative activity in animal models and humans.

3.6 Be more effective than GHK-CU on skin regeneration.

In veterinary studies, creams made from these new copper complexes produced rapid and scar-free healing in dogs after spaying operations, and in young horses after leg-straightening operations. This allowed the dogs to be returned to their owners in four days instead of the usual five, while the foals were returned in five days instead of seven.

In humans, four small, placebo-controlled studies were directed by the world-reknown dermatologist Howard Maibach, who reported that creams containing the new copper peptides produced faster skin healing after skin injuries induced by tape stripping, acetone burns (removal of skin lipids), 24-hour detergent irritation, and nickel allergy inflammation.

4.0 At Skin Biology, I feel that we have attained all the goals in items 3.1 to 3.6 but further testing is still needed. My goal is to develop the ultimate method of using copper peptides for skin remodeling and regeneration.

4.1 Over the past five years, we have been filing patent applications and developing this technology into useful products.

5.0 Projected future uses of copper peptides

GHK-Copper remains the best molecule for internal medical treatments. I predict that, in time, the newer breakdown resistant, highly adhesive copper peptides under development at Skin Biology should prove better for cosmetic and superficial uses such as post-procedure dermatological healing, and development of scarless surgical procedures.

It is possible that GHK-copper could be used clinically to protect and speed repair of damaged organs. H. Paul Ehrlich found that intra-muscular injection of  into the thigh muscle of rabbits raised circulating wound macrophages in the blood and accelerated the healing of distant wounds in the rabbit ear. Patients might be pre-treated with GHK- Copper before surgery to enhance post-surgical repair. Based on rabbit models, a dosage of 30 milligrams of  should suffice. The molecule is also very beneficial on kidney organ culture. Thus,  might be infused into patients with kidney failure to exert its tissue protective and repair actions.

The newer, second generation copper peptides produced at Skin Biology appear to be very useful for post-procedure recovery after skin peels, dermabrasion and laser resurfacing. The combination of hydroxy acids and these peptides slowly, over a period of several months, reduces old scars and skin lesions. This method is economical and avoids the complications that often occur after chemical peels or laser treatments. In experimental studies, the use of such types of copper peptides after surgical procedures often results in scarless or nearly scarless healing.

Why we list our ingredients the way we do:

1. Some confusion has arisen as to te way we list product ingredients.

2. FDA regulations require that we list the ingredients in the form that we purchase them.

3. For example, for CP Serum we list:

Items 90 and 130 - CP Serum
Purified water, hydroxyethylcellulose, copper-peptides (hydrolyzed soy protein plus copper chloride), allantoin, polysorbate-20, diazolinydinyl urea, methylparaben, propylparaben, vitamin E TPGS, bronopol.

4. We purchase copper salts and soy peptides and form the chelate as we mix them together in water. This is a trivial process that any high school chemistry student could do.

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For the slow, making copper peptides by the numbers

	Making GHK-copper (II)	Making Skin Biology's peptide-copper (II) complexes	If there are no peptides present
Step 1	Dissolve a copper salt such as copper chloride in water. This produced a slightly acidic solution.	Dissolve a copper salt such as copper chloride in water. This produced a slightly acidic solution.	Dissolve a copper salt such as copper chloride in water. This produced a slightly acidic solution.
Step 2	Dissolve into the solution at least equimolar peptide	Dissolve in at least equimolar peptide	No peptide is added
Step 3	Raise the pH of the solution to 5.5 to 7.5 as desired for final pH	Raise the pH of the solution to 5.5 to 7.5 as desired for final pH	Raise the pH of the solution to 5.5 to 7.5 as desired for final pH
Step 4	The GHK-copper (II) complex is formed	The soy peptide-copper (II) complex is formed	A white gelatinous precipitate of copper (II) hydroxide forms. Unchelated copper (II) will not stay in solution at physiological pHs.

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Dr. Loren Pickart has spent 40 years working on ways to keep you younger and is the discoverer of the tissue remodeling copper-peptides. He and his wife, Charlene, worked to start ProCyte Corporation in 1985 to develop his first generation of GHK-Copper products and Skin Biology in 1994 to create a second generation of more effective, skin remodeling copper peptides.

Products based on GHK-Copper are marketed by AdviCare, American Crew, Amuchina (Europe), Atelier Esthetique, Bard Medical, BioPharm (Middle East), Creative Nail Design, Johnson & Johnson, Schering AG, Neutrogena, Osmotics, ProCyte, Sigmacon Medical Products (Canada) and Tanox Biosystems (Asia). These include products for Cosmetic Skin Renewal: Neutrogena Visibly Firm Night Cream®, Neutrogena Visibly Firm Eye Cream®, Neutrogena Visibly Firm Moisture Makeup®, Neutrogena Visibly Firm Eye Treatment Concealer®, Neutrogena Visibly Face Lotion®, Neutrogena Visibly Firm Body Lotion®, Visibly Firm Face Lotion SPF 20®, Blue Copper Firming Elasticity Repair®, Climate Extreme Body Repair with Copper Peptide®, Simple Solutions Pure Copper Skincare products (Pure Copper Night Renewal®, Pure Copper Morning Dew®, Ultra Copper Firming Serum®, Pure Copper Eye Repair®), Blue Razor Aftershave®, Neova® Eye Therapy, Neova® Night Therapy Creme®, Neova® Day Therapy, Neova® Body Therapy Lotion®, Neova® Cuticle Therapy, Neova® Antioxidant Therapy Serum with GHK Copper Peptide Complex(TM),  Neova® Cuticle Therapy,  Neova® Therapy Cleansing Bar, Neova® Therapy Mattifying Serum, Neova® Therapy Copper Moisture Mask®, Neova® After Shave Therapy and Neova® Body Scrub,  for Veterinary Wound Healing: Iamin-Vet Skin Care Gel® and Iamin-Vet Wound Cleanser®. for Wound Healing: Iamin Gel Wound Dressing®, Iamin Impregnated Gauze Dressing®, Iamin Wet Dressing (copper-saline)®, Iamin-2 Hydrating Gel®, and Iamin Wound Cleanser®; for Hair Transplantation and improving the success of hair transplants: GraftCyte® Advanced Hair Restoration Technology: GraftCyte® Concentrated Spray, GraftCyte® Moist Dressings, GraftCyte® Hydrating Mist, GraftCyte® Post-Surgical Shampoo and Conditioner, and GraftCyte® Head Start Single Patient Pack; for Stimulation of Hair Growth and Hair Vitality: American Crew Revitalize Daily Shampoo®, American Crew Revitalize Spray Solution®, American Crew Revitalize Daily Conditioner®, American Crew Revitalizing Serum®, American Crew Revitalize Daily Moisture Shampoo®;  Tricomin® Solution Follicle Therapy Spray, Tricomin® Revitalizing Shampoo, Tricomin® Restructuring Conditioner and Tricomin® Conditioning Shampoo; and for Post-Surgical Skin Healing after laser resurfacing, dermabrasion, and chemical peels: Complex Cu3® Intensive Repair Cream, Complex Cu3® Hydrating Gel, Complex Cu3® Post Laser Lotion, and Complex Cu3® Gentle Face Cleanser.  In addition, human clinical studies of Pickart's inventions for bone healing and healing of intestinal irritations (Inflammatory Bowel Disease or Crohn's disease) have given positive results. He received his Ph.D. in Biochemistry from the University of California at San Francisco. For details on copper-peptide tissue remodeling - see: www.skinbiology.com/copperpeptideregeneration.html.

Skin Biology is developing Dr. Pickart's second generation of improved skin remodeling copper peptides into new skin remodeling products for uses such as restoring skin to a younger-looking condition, removing blemishes and scars, improving hair health, and helping skin recovery after lasers, peels, and dermabrasion.

Picture - Santorini, Greece - Dr. Pickart and his daughter Francoise.
 

Copyright 1999-2008 by Dr. Loren Pickart

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