This seems very promising!

Previous topic - Next topic

0 Members and 1 Guest are viewing this topic.

MJohn

This has got me super excited!  I haven't had any luck finding the actual study listing the 22 compounds and would love some assistance gentlemen.  In the video below, Marcus Maximillian EAU17 e-Poster 2nd Prize Non-Oncology winner discusses his new findings for peyronies.  He mentions that part of the reason that there are no viable oral treatments is that they don't have solid evidence as to what drugs affect the cells of the penis.  His team developed a method of quickly and easily identifying drugs/compounds that do affect the actual cells of the penis.  The short blurb I was able to find on the Internet states they have identified 22 compounds but I have not been successful in finding what they are. I also want to know if they are continuing this to find more drugs/compounds.  

https://www.youtube.com/watch?time_continue=5&v=X_zhzaQIbfQ




Arabia

Good find and the best thing is he is  so young that he doesn't even have any papers to his name yet. Young mind, curious, and big clinical problem is a good combination.

That is a poster session so there will be no paper and you can be sure they are not going to reveal their drug list at this point in time given the potential large financial rewards for developing a possible oral treatment for Peyronie's.   Problem is that from his lab bench to an approved clinical product, if some of those drugs do show promise, will take at least five years even in Europe.

Prize Winners – EAU17 London  (to the link police the Euro Assoc. of Urology is a professional organization and not a commercial entity)

NeoV

There are plenty of compounds that inhibit tgf-beta in the penis. Way more than one would imagine. As for drugs, we certainly need more. Sounds good.

pey ron

I've started to look into a few compounds that inhibit tgf-beta1.

Just off the top of my mind (I might forget some):

- sulforaphane (from broccoli extract)
- ursolic acid (from holy basil)
- lithium chloride
- curcumin (from turmeric)
- quercetin
- ginsan (from panax ginseng)

If you guys can add to the list, it would be great!

EDIT: forgot ursolic acid
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **

Paolo

Hi pey ron, MSM
Methylsulfonylmethane MSM is the primary metabolite of DMSO in humans, and is also a metabolite of sulfur-containing amino acids  :)

I take it with tiny amount (15mg) of Vitamin C powder 3-4 times a day.
Whenever you find yourself on the side of the majority, it is time to pause and reflect.

skunkworks

What is the reasoning behind the MSM, does it inhibit tgf-beta in the penis?
This is an emotionally destructive condition, we all have it, let's be nice to each other.

Review of current treatment options by Levine and Sherer]

Paolo

Not sure, it's just that pey ron mentioned sulforaphane, MSM contains it, thats all  :-\

see Inhibitory Effects of Methylsulfonylmethane on Ventricular Hypertrophy Related Gene Expression

I take it specifically for  anti-inflammatory actions  :)
Whenever you find yourself on the side of the majority, it is time to pause and reflect.

pey ron

@Paolo...

MSM is Methylsulfonylmethane, chemical formula C2H6O2S.

Sulforaphane's chemical formula is instead C6H11NOS2.

Both of them refer to exactly one specific molecule. There's no way one can "contain" the other one.
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **

Paolo

Thanks pey ron, that is noted  :)

I am going to look into Ursolic acid as it appears to increase 'brown' fat  :)
Whenever you find yourself on the side of the majority, it is time to pause and reflect.

pey ron

@paolo: if you find a good source for ursolic acid (or for any of the above), please share!
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **

Paolo

Hi pey ron, I am trying Swanson holy basil leaf (tulsi) caps at present (they contain standardized for 2% ursolic acid), the price seemed very reasonable for 120 caps (30 day supply).  :)

If I feel any benefits I may look for more with a higher concentration of UA  :)

Amongst other benefits Ursolic Acid Increases Skeletal Muscle and Brown Fat and Decreases Diet-Induced Obesity, Glucose Intolerance and Fatty Liver Disease https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3379974/

I exercise quite a bit so the increased skeletal muscle (at my age) would be very welcome  :)

For those who have/had alcohol issues UA is apparently of benefit also in respect to the heart  :-\
Whenever you find yourself on the side of the majority, it is time to pause and reflect.

pey ron

@Paolo, I was thinking of a bulk source, like this one:

http://www.bulksupplements.com/holy-basil.html

except I don't trust that company much...
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **

Paolo

Not sure pey ron, looks reputable  :-\

I may also try Tulsi tea and see also how that goes.

Anyone trying to conceive should avoid Ursolic acid as it has a negative effect on sperm motility, I am curious on the effect Holy basil leaf has on testosterone.

I have PM you with alternative to one chosen, hope it is okay  :)
Whenever you find yourself on the side of the majority, it is time to pause and reflect.

MJohn

I bought EGCG extract.  This also seems to be potent TGF-B inhibitor and has other health benefits.  Was cheap too.  

pey ron

Quote from: NeoV on March 27, 2017, 12:45:32 AM
plenty of compounds that inhibit tgf-beta in the penis

@NeoV: can you please share those that you know of?
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **


pey ron


-----8<----[ 10.1016/j.jsxm.2016.02.018 ]-------

DEVELOPMENT OF A HIGH-THROUGHPUT, CELLBASED ASSAY FOR ANTI-MYOFIBROBLAST ACTIVITY IN PEYRONIE'S DISEASE

Mateus, M.1, *;
Stebbeds, W.2;
Ameyaw, B.2;
Raheem, A.3;
Spilotros, M.3;
Garraffa, G.3;
Muneer, A.3;
Christopher, N.3;
Cellek, S.1;
Ralph, D.3

1 Anglia Ruskin University, United Kingdom;
2 Cranfield University, United Kingdom;
3 University College London Hospital, United Kingdom

Objectives:

Peyronie's disease (Peyronies Disease) is a fibrotic disorder characterised by the formation of plaques in the tunica albuginea (TA) of the penis, which can result in curvature, pain and erectile dysfunction. Peyronies Disease has been affecting a growing number of men worldwide; however, no method is available to quickly and inexpensively test a large number of potential new drugs, known as high throughput screening (HTS). The aim of this study was to develop and validate an HTS assay for the identification of compounds with anti-myofibroblast activity in cells established from tunica albuginea.

Methods:

Fibroblasts cultures established from the TA of patients with and without Peyronies Disease were exposed to TGF-b1 in order to differentiate them into myofibroblasts. Alpha-smooth muscle actin (a-SMA) immunostaining was assessed using immunohistochemistry (IHC) and immunocytochemistry (ICC). The a-SMA mRNA levels were measured using real-time RT-PCR (RT-qPCR). In Cell Western (ICW) method was used to develop the HTS assay measuring a-SMA staining and cell numbers, in cells isolated from non-Peyronies Disease tissue and exposed to control conditions, TGF-b1 and 22 FDA approved drugs with potential anti-myofibroblast activity.

Results:

IHC, ICC and RT-qPCR showed higher a-SMApositive cells or a-SMA expression in fibroblasts derived from Peyronies Disease plaque tissue than from cells isolated from non-Peyronies Disease tissues. The ICW assay was able to effectively and reproducibly detect TGF-b1-induced myofibroblast transformation, obtaining an average Z' of 0.84 with a CV of 6% in 96 well plates. Using the ICW assay, 5 of the 22 drugs were found to significantly inhibit TGF-b1-induced myofibroblast transformation. Full concentration response curves were constructed for all the five compounds, where an inverted sigmoid curve with an upper and lower plateau was observed and IC50 values could be calculated. DNA staining showed that the cell numbers were not significantly reduced by any of the five compounds.

Conclusions:

The first assay amenable to HTS was developed for the detection of compounds with anti-myofibroblast activity in cells isolated from human non-Peyronies Disease tissue. The data presented herein suggests that this novel assay can potentially be used for identification of novel compounds or repositioning of approved drugs not only for Peyronies Disease but also for other fibrotic disorders.

Funding:

Partly funded by ESSM Research Grant to W.J.S.

Disclosure:

Work supported by industry: no.

-----8<----[ 10.1016/S1569-9056(17)31161-2 ]-------

Development and validation of a phenotypic high-throughput, cell-based assay for anti-myofibroblast activity in Peyronie's disease

Ilg M.M.1,
Mateus M.1,
Stebbeds W.2,
Ameyaw B.2,
Raheem A.3,
Spilotros M.3,
Capece M.3,
Parnham A.3,
Garaffa G.3,
Christopher N.3,
Muneer A.3,
Cellek S.1,
Ralph D.3

1 Anglia Ruskin University, Faculty of Medical Science, Chelmsford, United Kingdom,
2 Cranfield University, Cranfield Health, Bedfordshire, United Kingdom,
3 University College London Hospital, Dept. of Andrology, London, United Kingdom

INTRODUCTION & OBJECTIVES:

Peyronie's disease (Peyronies Disease) is characterized by the formation of a fibrotic plaque in the penile tunica albuginea (TA) associated with penile curvature, pain and erectile dysfunction. Currently the medical interventions are limited, with no clear evidence. This study aimed to develop and validate a novel phenotypic high-throughput screening (HTS) assay allowing rapid and inexpensive testing of several compounds for their anti-myofibroblast activity in cells derived from human
TA, as well as, developing secondary functional screening assays to confirm the anti-myofibroblast activity.

MATERIAL & METHODS:

Human primary fibroblasts were isolated from TA of patients with Peyronies Disease. The cells were stained for vimentin and desmin using immunofluorescence. In Cell Western (ICW) was used to perform the HTS assay measuring alpha smooth muscle actin (α-SMA) staining and cell numbers where the cells were exposed to TGF-β1 to induce myofibroblast transformation. A modified ICW was used to measure intra- and extracellular type V collagen (Col V) staining. Fibroblast-populated collagen lattices (FPCLs) were used to measure the contractile function of the cells. MTT assay was performed to assess cell viability. The protein expression results were also confirmed with mRNA expression using RTPCR.

RESULTS:

The cells obtained from TA tissue were vimentin positive and desmin negative, confirming their fibroblast identity. The ICW method was able to reproducibly and effectively detect TGF-β1-induced myofibroblast transformation (Z'=0.89; CV=6%) in 96 well plates. Five FDA approved drugs were tested and SB-505124, a selective inhibitor of TGF-β1 receptor, used as a positive control, significantly inhibited TGF-β1-induced myofibroblast transformation. For each compound a full concentration response curve was constructed and IC50 values were obtained ranging from 0.03 µM to 24 µM. ICW for extra- and intracellular Col V showed that two of the drugs tested reduced TGF-β1 induced Col V expression in a concentration dependent manner. FPCL assay showed that the drugs were capable of decreasing contraction of collagen lattices. MTT assay showed that the drugs did not affect cell viability in the used concentrations.

CONCLUSIONS:

The fibroblast identity of our cells was confirmed by vimentin and desmin staining. Using SB-505124 as a positive control we were further able to validate our HTS assay which identified five FDA approved drugs to have anti-myofibroblast activity. The secondary functional assays have further confirmed the results from the primary screen. This battery of assays can be used in the future to screen further compounds for the development of new medicines for fibroproliferative diseases such as Peyronies Disease.

Acknowledgement:

This project has partly been funded by European Society of Sexual Medicine.

-----8<----[ 10.1016/j.jsxm.2017.03.024 ]-------

DEVELOPMENT OF SECONDARY ASSAYS TO VALIDATE HITS FROM PRIMARY PHENOTYPIC SCREEN FOR ANTI-MYOFIBROBLAST ACTIVITY IN PEYRONIE'S DISEASE

Ilg, M.1;
Mateus, M.2;
Stebbeds, W.3;
Raheem, A.4;
Capece, M.4;
Parnham, A.4;
Garaffa, G.4;
Muneer, A.4;
Christopher, N.4;
Cellek, S.2;
Ralph, D.4

1 Anglia Ruskin University, Faculty of Medical Science, Chelmsford, United Kingdom;
2 Anglia Ruskin University, Chelmsford, United Kingdom;
3 Cranfield University, Bedfordshire, United Kingdom;
4 University College London Hospital, London, United Kingdom

Objective:

Peyronie's disease (Peyronies Disease) is a fibrotic disorder characterised by the formation of localised fibrous plaques in the tunica albuginea (TA) of the penis, which can result in pain during erection, deformities and erectile dysfunction. Peyronies Disease has been affecting a growing number of men worldwide, however medical treatment for Peyronies Disease is currently lacking efficiency. Following the validation of our primary high-throughput screen for the detection of compounds with anti-myofibroblast activity in cells established from TA, this study aims to develop secondary screening assays to confirm the antimyofibroblast activity.

Methods:

Fibroblasts derived from patients with and without Peyronies Disease were exposed to TGF-b1 to induce myofibroblast transformation and cells were stained for various types of collagen and fibronectin. A modified In Cell Western (ICW) was used to measure intra- and extracellular type V collagen (Col V) staining and cell numbers, in non-Peyronies Disease TA cells exposed to control conditions, TGF-b1 and two FDA approved drugs. Fibroblast-populated collagen lattices (FPCLs) were used to measure differences in contraction of fibroblasts treated with drugs and under control conditions. MTT assay was performed to show the effects of the drugs on cell viability.

Results:

Cells treated with TGF-b1 showed an increase in Col V expression. Preliminary results of ICW for extra- and intracellular Col V showed that the two drugs significantly reduced TGF-b1 induced Col V expression in a concentration dependent manner. Preliminary results for FPCL assay show that the drugs are capable of decreasing contraction of collagen lattices. Preliminary results for MTT assay suggest that the drugs do not affect cell viability in the used concentrations.

Conclusion:

To complement the results of our previously described phenotypic screening assay, we have developed secondary screening assays targeting collagen production, contraction and cell viability. The preliminary data suggests that these novel assays have the potential to help confirming the anti-myofibroblast activity of drugs identified by our primary phenotypic screen.

Policy of full disclosure:

None

-----8<----[ 10.1016/j.jsxm.2017.03.098 ]-------

FURTHER VALIDATION OF PHENOTYPIC HIGH-THROUGHPUT, CELL-BASED ASSAY FOR ANTI-MYOFIBROBLAST ACTIVITY IN PEYRONIE'S DISEASE

Ilg, M.1;
Mateus, M.2;
Stebbeds, W.3;
Raheem, A.4;
Capece, M.4;
Paranham, A.4;
Garaffa, G.4;
Muneer, A.4;
Christopher, N.4;
Cellek, S.2;
Ralph, D.4

1 Anglia Ruskin University, Faculty of Medical Science, Cambridge, United Kingdom;
2 Anglia Ruskin University, Cambridge, United Kingdom;
3 Cranfield University, Bedfordshire, United Kingdom;
4 University College London Hospitals, London, United Kingdom

Objective:

In Peyronie's disease, characterized by formation of a fibrotic plaque in the penile tunica albuginea (TA) leading to curvature, pain and erectile dysfunction, the current medical treatment is nearly limited to surgery, despite a growing number of men worldwide are affected. The aim of this study was to further validate our previously developed phenotypic high-throughput screening (HTS) assay that allows quick and inexpensive testing of numerous compounds for their anti-myofibroblast activity in cells established from TA.

Methods:

Cells were stained for vimentin and desmin. Fibroblasts derived from patients with and without Peyronies Disease were exposed to TGF-b1 to induce myofibroblast transformation. In Cell Western (ICW) was used to perform the HTS assay measuring a-SMA staining and cell numbers, in cells exposed to control conditions, TGF-b1, FDA approved drugs and SB-505124, a selective inhibitor of type I TGF-b receptor that has been reported to inhibit TGF-b1-induced myofibroblast differentiation.

Results:

Our cells were shown to be vimentin positive and desmin negative, confirming fibroblast identity. The ICW method was able to reproducibly and effectively detect TGF-b1-induced myofibroblast transformation, obtaining an average Z' of 0.89 with a CV of 6% in 96 well plates. 5 of the FDA approved drugs tested and SB-505124, used as a positive control validating our assay, significantly inhibited TGF-b1-induced myofibroblast transformation. For each compound a full concentration response curve (CRC) was constructed, where an inverted sigmoid curve with an upper and lower plateau was observed and IC50 values could be calculated.

Conclusion:

Our previously described phenotypic screening assay could be further validated to screen for compounds to prevent TGF-b1-induced myofibroblast transformation. The fibroblast identity of our cells was confirmed by vimentin and desmin staining. The use of SB-505124 as positive control could further validate our assay that identified 5 of the FDA approved drugs to have anti-myofibroblast activity.

Policy of full disclosure:

None

-----8<--------------------------
Please go to PROFILE then FORUM PROFILE to replace this signature line text with your profile info such as
age, date of onset, symptoms, treatments tried,
relationship status, etc
** You will waste less time and get better answers **