IGF-1 frequency dependency of effectiveness

[vkg1]

Let's say you have 1mg of IGF1LR3 and that's all you'll ever have. And let's say you are going for localized growth in just a couple places like chest and calves or something (ie not going to be using "systemically").

How much does making the most of that 1mg of IGF depend on how often you use it? For example, would you theoretically get the same final total effect if you were to inject once per week as 3-4 times per week (just at 1/3-1/4 the rate of development)?

Also, what if you were to inject 3-4 times a week for two weeks and then take two months off, then another two weeks on followed by two months off? Would this be "wasting" IGF relative to a protocol with closer to the minimum-to-avoid-receptor-downgrades schedule of injections?

Is the total amount injected, independent of time, the primary determinant of results (all other variables but frequency equal), or do you have to maintain a certain frequency and regularity to get good results?

 

[CryingEmo]

I feel that every 3 or 4 days at a higher dose is better than ED or EOD at a smaller dose.

 

[pumbertot]

Let's say you have 1mg of IGF1LR3 and that's all you'll ever have. And let's say you are going for localized growth in just a couple places like chest and calves or something (ie not going to be using "systemically").

How much does making the most of that 1mg of IGF depend on how often you use it? For example, would you theoretically get the same final total effect if you were to inject once per week as 3-4 times per week (just at 1/3-1/4 the rate of development)?

Also, what if you were to inject 3-4 times a week for two weeks and then take two months off, then another two weeks on followed by two months off? Would this be "wasting" IGF relative to a protocol with closer to the minimum-to-avoid-receptor-downgrades schedule of injections?

Is the total amount injected, independent of time, the primary determinant of results (all other variables but frequency equal), or do you have to maintain a certain frequency and regularity to get good results?

yes what CryingEmo says basically is true up to around 50-60mcgs per dose.

if we think about the effect we want from the igf and that is localised grwoth then yes it is true that you will achieve the same level of hyperplasia even if spreading over a greater period of time as long as the dose is large enough to force the muscle into hyperplasia in the first place.
so you would want to use at least 20mcg for this purpose.

if you only have access to 1mg ever I would say that you would want to target only one muscle group but you could do 2 at a stretch. and hit each of those with 20mcg each per week so you will get 25 weeks out of that 1mg.
but if you can stretch to 2mg of igf for those 25 weeks then much better hitting each with 40mcg one per week.

of course this is far from ideal but it will cause hyperplasia which is what we want afterall.

 

[KurtisWicked]

Not sure if I should make another thread for this but figure its along the lines of IGF-1 effectiveness. What would be the best routine to maximize the effects of IGF-1? Low reps, heavy weight or high reps light weight? I'm just curious as to if there's a difference.

Sorry for the highjack.

 

[TripDog]

I feel that every 3 or 4 days at a higher dose is better than ED or EOD at a smaller dose.

I like smaller doses. IMO people just waste the stuff with high doses.

 

[pumbertot]

Not sure if I should make another thread for this but figure its along the lines of IGF-1 effectiveness. What would be the best routine to maximize the effects of IGF-1? Low reps, heavy weight or high reps light weight? I'm just curious as to if there's a difference.

Sorry for the highjack.

i think a combination would be best. just dont overdo the low reps for too long a period of time, cycle for 8 weeks etc.

but you can do both in the same workout which works really well. after warming up 2 sets of 6-8 reps. then last set do a drop set,increase weight for 4 reps to failure, then immediately drop so you can manage another 4 and so on until you get 20-30 reps total. best of both worlds. just dont do those heavy 4 reppers for more than 8 weeks or so at a time to avoid the risk of tears(from experience )

 

[CryingEmo]

I like smaller doses. IMO people just waste the stuff with high doses.

I guess it depends on how much your body can use at a time.


I think the jury is still out on that one.


I have more vascularity and fullness for longer with a larger dose.

 

[Bobaslaw]

I guess it depends on how much your body can use at a time.


I think the jury is still out on that one.


I have more vascularity and fullness for longer with a larger dose.

Pumps and vascularity are not effects that should be used to judge the effective dose of IGF-1 as these are just transitory effects and have nothing to do with the only real permanent purpose it is legitimately used for: hyperplasia. This cannot be judged by "feeling" in the least. If you want good pumps there are plenty of cheaper products for this purpose...

Sure, you can have fun with the Pumps and Vascularity at higher doses(its a great feeling I agree) but only for a shorter time before your receptors are quickly downregulated, thus sacrificing the real maximum potential of IGF for hyperplasia.

Just my 2 cents

 

[pumbertot]

Pumps and vascularity are not effects that should be used to judge the effective dose of IGF-1 as these are just transitory effects and have nothing to do with the only real permanent purpose it is legitimately used for: hyperplasia. This cannot be judged by "feeling" in the least. If you want good pumps there are plenty of cheaper products for this purpose...

Sure, you can have fun with the Pumps and Vascularity at higher doses(its a great feeling I agree) but only for a shorter time before your receptors are quickly downregulated, thus sacrificing the real maximum potential of IGF for hyperplasia.

Just my 2 cents

yes i agree with this statement and thats why call 60mcg a high dose, the highest you would want for hyperplasia imo.

 

[pistonpump]

yes i agree with this statement and thats why call 60mcg a high dose, the highest you would want for hyperplasia imo.

60mcg in each muscle or is that the total before splitting? just want to make it clear what you are saying...

 

[pumbertot]

60mcg in each muscle or is that the total before splitting? just want to make it clear what you are saying...

50-60mcg split. taking it each might eventually lead to coleman guts.

 

[TripDog]

50-60mcg split. taking it each might eventually lead to coleman guts.

I'm on my 7th vial and no issues at all, I always kept doses low.

 

[comacho]

what are your shooting regimen Trip?

just curious as i hear all kinds of different dosages/duration

 

[Newbie40plus]

what are your shooting regimen Trip?

just curious as i hear all kinds of different dosages/duration

I would like to know this too. I just started with LR3 IGF for the first time yesterday.

 

[papapumpsd]

I'm on my 7th vial and no issues at all, I always kept doses low.

Trip, how much time is takin' off between vials (1mg vials I'm assuming)??

 

[datBtrue]

THE most important point to keep in mind is this:

IF you are low dosing it and using for more than say 30 days...and you were talking about using it for 2-3 months...you need to understand that your reconstituted vial WILL lose potency...particulary after 30 days...and may be of zero effect after 60 days.

What can you do?

Reconstitute.
Draw out a third & inject it into a sterile vial.
Draw out another third & inject it into another sterile vial.
Freeze these "new" vials & keep them frozen until time of use.
Place the remaining 3rd in the refrig. and use that first.

Thats probably the best you're going to be able to do.

Any dosing protocol is of no use if the compound has become impotent. Keep that in mind first.

 

[TripDog]

Trip, how much time is takin' off between vials (1mg vials I'm assuming)??

Doses and use was all over the place...didn't ever run a straight 4 week cycle.

 

[papapumpsd]

THE most important point to keep in mind is this:

IF you are low dosing it and using for more than say 30 days...and you were talking about using it for 2-3 months...you need to understand that your reconstituted vial WILL lose potency...particulary after 30 days...and may be of zero effect after 60 days.

What can you do?

Reconstitute.
Draw out a third & inject it into a sterile vial.
Draw out another third & inject it into another sterile vial.
Freeze these "new" vials & keep them frozen until time of use.
Place the remaining 3rd in the refrig. and use that first.

Thats probably the best you're going to be able to do.

Any dosing protocol is of no use if the compound has become impotent. Keep that in mind first.

Dat, I have read repeatedly that IGF-1 is stable (retains full potency) at RT for many months if reconned in 0.6% AA.

I see you used the phrase "...lose potency..." which implies it will still have activity, just not the original 100%. If this is what you're referring to, please advise.

 

[Bobaslaw]

Dat, I have read repeatedly that IGF-1 is stable (retains full potency) at RT for many months if reconned in 0.6% AA.

I see you used the phrase "...lose potency..." which implies it will still have activity, just not the original 100%. If this is what you're referring to, please advise.

Papa, it really depends whether you trust this Gropep Study:

Storage of LR3 IGF-1
*Study conducted by Gropep
*posted by LakeMounted @ *************.com

The stability of a liquid solution of LR3IGF-I was monitored for a period of two years at storage conditions of -20 C, +4 C, +22 C, and +37 C. The final concentration of LR3IGF-I was in acetic acid. At various time points, samples were taken and compared to a lyophilized control (stored at 4 C). Listed below are the stability results for each respective storage condition.

Storage Condition: -20 C (-4 F)
Biological Potency No Change up to 2 years
Immunological Activity No Change up to 2 years
Mobility of Protein No Change up to 2 years
Elution Profile by reversed phased HPLC No Change up to 2 years

Storage Condition: +4 C (39.2 F)
Biological Potency No Change up to 2 years
Immunological Activity No Change up to 2 years
Mobility of Protein No Change up to 2 years
Elution Profile by reversed phased HPLC No Change up to 2 years

Storage Condition: +22 C (71.6 F)
Biological Potency No Change up to 2 years
Immunological Activity No Change up to 2 years
Mobility of Protein No Change up to 2 years
Elution Profile by reversed phased HPLC No Change up to 2 years

Storage Condition: +37 C (98.6 F)
Biological Potency No Change up to 1 year
Immunological Activity No Change up to 1 year
Mobility of Protein No Change up to 1 year
Elution Profile by reversed phased HPLC No Change up to 1 year

 

[datBtrue]

Dat, I have read repeatedly that IGF-1 is stable (retains full potency) at RT for many months if reconned in 0.6% AA.

I see you used the phrase "...lose potency..." which implies it will still have activity, just not the original 100%. If this is what you're referring to, please advise.

The Acetic Acid is used to maintain the PH of the solution and thus reduce protonation & deprotonation of the carboxyl & amino groups of the amino acids in the peptide chain all of which have different isoelectric points at which "equilibrium" is achieved. The PH of the solution is a compromise for the entire chain. This small degradation effect is cumulative but minimal...

While PH may address in part deamidation (temperature playing a role as well), it does not negate other degradation pathways such as oxidation and aggregation

Based on my communication with the biotechnology firm that I will employee to sythesize my peptides specifically on the topic of storage I believe my statements to be true.

Based on my own use of dosages of IGF-1 LR3 out of the same reconstituted vial for a period of 3 months wherein I experienced a decrease in insulin-like effects...so I believe my statements to be true.

Based on my reading of the book "Therapeutic Peptides and Proteins" part of which is quoted below...I believe my statements to be true:

Lyophilized powders can be quite stable as long as they are not reconstituted. For example, Activase® lyophilized powder shows no significant loss of bioactivity after storage for more than 4 years at controlled room temperature. Degradation in the solid state often takes place by aggregation. The stability of a protein in the solid state is dependent on the moisture content of the solid, temperature, and composition of the formulation. Stability of proteins in the solid state on exposure to moisture or elevated temperatures is very important because such conditions are frequently encountered in pharmaceutical systems.
...
Currently, most recombinant proteins are developed as parenteral dosage forms, and this trend is likely to continue, at least in the immediate future. The hospital pharmacist will be closely associated with such therapy. About half of the parenteral products are marketed as lyophilized powders, which need to be reconstituted just before use. The diluent is usually WFI USP and may be provided with the dry powder as a convenience. Products such as Humatrope® (hGH) supply their own diluting solution, which in this case contains 0.3% m-cresol as a preservative and 1.7% glycerin as a stabilizer and tonicity agent. Some products are to be reconstituted with bacteriostatic water for injection, which contains benzyl alcohol as a preservative. These products can generally be stored for about 14 days after reconstitution. For products that do not contain any antimicrobial preservatives, the manufacturer will generally specify that the product be used immediately after reconstitution to minimize the possibility of compromised sterility.
​

You know what I'm trying to figure out is really the effect of oxidation. If I have 1000mg of a simple peptide made in bulk and sealed in a single vial...what is the effect of me breaking the seal and placing 5mg in single vials or containers and then storing 200 tiny vials containing 5mg each.

It isn't practical for me to be able to create a vaccum in each tiny vial.

I don't know...in practical terms...but I will find out. Unfortunately my source of information is likely to be overly cautious.

 

[datBtrue]

The following is from Imgenex Corp. (develops novel reagents for scientific study)

http://www.imgenex.com/recomb_tds.php?id=1049

Recombinant Human Insulin Like Growth Factor-I (IGF-1)

Stability

Lyophilized IGF-1 although stable at room temperature for 3 weeks, should be stored desiccated below -18 C. Upon reconstitution IGF-I should be stored at 4 C between 2-7 days and for future use below -18 C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please avoid freeze-thaw cycles.​

 

[datBtrue]

These sections of the following guide helped clarify a few questions I had. The following is from

A GUIDE TO HANDLING AND STORING PEPTIDES
Mimotopes Peptides & Immunology​

Storage of Dry Peptide​

When peptides are received, ensure they are kept in a cool, dark place. For best preservation, store them under refrigeration at 4°C or colder, away from bright light. Dry peptides are stable at room temperature for days to weeks, but for long term storage, -20°C is preferred. Contamination with moisture will greatly decrease long term stability of solid peptides. Each time you use some of the peptide, remove the container from cold storage and allow it to equilibrate to room temperature or slightly warmer before opening it. This will reduce the uptake of moisture from the air onto the cold surface of the solid peptide or the inside of the container. After removing the required quantity, re-seal the container, preferably under an atmosphere of dry nitrogen. This can be achieved by blowing a gentle stream of dry nitrogen into a plastic bag housing the container, taking great care to avoid blowing the peptide powder right out of the container. After the air is displaced, quickly cap the container, and then return it to cold storage. This procedure will minimize the oxidation of air-sensitive peptides as discussed later.

Storage of Peptide Solutions​

The shelf life of peptide solutions is limited, especially for peptides containing cysteine, methionine, tryptophan, asparagine and glutamine. To prolong the storage life of peptides in solution, use sterile buffers around pH5 to 6, and freeze aliquots for storage at -20°C or colder whenever possible. Avoid the use of frost-free freezers, which vary enormously in temperature during the frequent defrosting cycles. Repeated freeze-thaw cycles can also damage peptides.

Oxidation​

A characteristic of cysteine- and methionine-containing peptides is the tendency of these residues to oxidise. Susceptibility to oxidation is sequence-dependent and sometimes even minimal exposure to air of peptides containing these amino acids can lead to oxidation. If you wish to avoid oxidation, always work with degassed or deoxygenated solvents and solutions. If possible, maintain peptide solutions at acidic pH (<7). Rate of oxidation increases with pH, so even if the peptide is in the fully reduced form initially, some oxidation will occur if the peptide is maintained under neutral or basic conditions.​

 

[Bobaslaw]

Great info Dat!

I wanted to add some info I have found in my research as to IGF and the proper ph of its solvent.
IGF-1LR3 has a total positive charge and is deemed a "basic" peptide. This is determined by adding the charges of its acidic and basic residues. Acidic Residues are Asp(D), Glu(E), and the C Terminal -COOH. Basic residues are Arg (R), Lys(K), His(H) and the N Terminal -NH2.
Basic peptides ideally require an acidic solvent in which to be properly reconstituded and remain most stable.

Also, an observation on the significance of oxidation as it pertains to IGF-1. Cysteine and Methionine formed disulfide bonds are both very succeptible to ph based oxidation, and reduction for that matter. Particularly Cys with respect to the IGF chain, is responsible for the three intra-chain disulfide bonds (Cys-Cys) giving the peptide its unique 3D folded shape. Cys-Cys disulfide bonds are shown in the image (C-C):

These disulfide bonds and thus the folded shape of IGF are directly responsible for it's biological activity. A study on IGF-1 and its disulfide bonds shows that it has full biological activity when all 3 disulfide bonds are intact. Biological activity decreases with IGF-1 "intermediates" in which fewer, or incorrectly paired, or missing disulfide bonds are exhibited:

Role of native disulfide bonds in the structure and activity of insulin-like growth factor 1: genetic models of protein-folding intermediates.

Narhi LO, Hua QX, Arakawa T, Fox GM, Tsai L, Rosenfeld R, Holst P, Miller JA, Weiss MA.
Amgen Inc., Amgen Center, Thousand Oaks, California 91320.

Insulin and insulin-related proteins contain three motif-specific disulfide bonds. Here we examine the role of these disulfide bonds in the folding and function of one family member, human insulin-like growth factor 1 (IGF-1). Analogues containing pariwise Cys-->Ser or Cys-->Ala substitutions were expressed in Escherichia coli, purified, and analyzed with respect to receptor-binding, solution structure, and thermodynamic stability. An analogue lacking all three disulfide bonds (designated des-Cys-IGF-1) is inactive and unfolded. Introduction of the [18-61] disulfide bond, previously shown to occur in an early intermediate in oxidative refolding [Miller, J. A., Owers-Narhi, L., Hua, Q. X., Rosenfeld, R., Arakawa, T., Rohde, M., Prestrelski, S., Lauren, S., S. Stoney, K. S., Tsai, L., & Weiss, M. A. (1993) Biochemistry (preceding paper in this issue)], results in a compact partially folded state with low but significant biological activity. Additional but incomplete structural organization and biological activity are observed following introduction of either the [6-48] or the [47-52] disulfide bonds. Native function, structure, and stability require the presence of all three disulfide bonds. These analogues provide genetic models of IGF-1 protein-folding intermediates. Their characterization suggests that bifurcation of the IGF-1 folding pathway reflects alternative late steps in the folding of a molten-globule intermediate.

 

[Royd The Noyd]

Each time you use some of the peptide, remove the container from cold storage and allow it to equilibrate to room temperature or slightly warmer before opening it. This will reduce the uptake of moisture from the air onto the cold surface of the solid peptide or the inside of the container.

I wonder if I could just draw from the vial in the fridge. I dont see any issues with that and I could avoid having to set it out to reach room temp each time.

 

[datBtrue]

Great info Dat!

I wanted to add some info I have found in my research as to IGF and the proper ph of its solvent.
IGF-1LR3 has a total positive charge and is deemed a "basic" peptide. This is determined by adding the charges of its acidic and basic residues. Acidic Residues are Asp(D), Glu(E), and the C Terminal -COOH. Basic residues are Arg (R), Lys(K), His(H) and the N Terminal -NH2.
Basic peptides ideally require an acidic solvent in which to be properly reconstituded and remain most stable.

I believe though that IGF-1 can be made with an "additive" when it is lypholized (i.e. freeze-dried) that will create the proper pH when the powder is rehydrated/reconstituted. So in that case a mild solvent or a weak concentration of hydrochloric acid would just be needed to dissolve the peptide as the proper pH environment is provided in the powder.

But the Chinese generic stuff isn't made this way. There is no additive. So we need to use a solvent that both dissolves and provides a proper pH environment...enter diluted Acetic Acid which performs both these duties.

This was my understanding of he current situation.

Also, an observation on the significance of oxidation as it pertains to IGF-1. Cysteine and Methionine formed disulfide bonds are both very succeptible to ph based oxidation, and reduction for that matter. Particularly Cys with respect to the IGF chain, is responsible for the three intra-chain disulfide bonds (Cys-Cys) giving the peptide its unique 3D folded shape. Cys-Cys disulfide bonds are shown in the image (C-C):

Awesome visual image & explanation Bob. THAT is how IGF-1 differs from my simple little GHRPs. Now I'm no longer worried about oxidation with regard to my GHRPs that lack those amino acids. I could buy a tub and split it up and freeze it but I don't have to break it down to tiny vials. I can take some out as needed and reconstitute in a sterile environment. Cool!

These disulfide bonds and thus the folded shape of IGF are directly responsible for it's biological activity. A study on IGF-1 and its disulfide bonds shows that it has full biological activity when all 3 disulfide bonds are intact. Biological activity decreases with IGF-1 "intermediates" in which fewer, or incorrectly paired, or missing disulfide bonds are exhibited:

From that study it appears that if you get the "wrong" kind of damage to IGF-1 it doesn't have the effect of reducing potency but rather eliminating its full biological value.

Awesome stuff Bob!

 

[datBtrue]

I wonder if I could just draw from the vial in the fridge. I dont see any issues with that and I could avoid having to set it out to reach room temp each time.

That quote comes from discussion of storage of the dry lypholized powder...in unreconstituted form. If you pull that unreconstituted powder out of the freezer it is best to let it get closer to room temp. before opening to withdraw a portion of the powder. Otherwise the disequilibrium is likely to draw moisture from the air into the lypholized powder. Moisture is not good. We are concerned about the moisture in the powder when we go back to refreeze it.

The admonition to avoid repeated freeze-thaw cycles primarily pertains to reconstituted solution. So if you reconstitute and freeze...than only thaw out when you are ready to use and never refreeze it.