? For fellow BATmen on daro and androgen stimulation. Great Article

[KocoPr]

I came across this 2023 article and i think it poses a great outcome for us who are cycling high T and darolutamide.

Am I correct that this article correlates positively with our BAT with Daro?

I find this article extremely fascinating although it is complex. I need to learn more about proteonomic or onomics for that matter and its effects on transcription.

We report for the first time the impact of the newly approved AR antagonist darolutamide using a very sensitive, label-free, global mass spectrometry–based approach and show a much better concordance between the regulation of protein and mRNA levels than found in previous studies with other compounds. Importantly, darolutamide is structurally different from other second-generation AR antagonists, such as enzalutamide and apalutamide. It is characterized by a flexible polar-substituted pyrazole structure, which leads to unique interactions within the AR ligand-binding pocket and to differential recruitment of cofactors compared to other AR antagonists [16], and to differences in the downstream regulated genes [62].

Comparative Proteomic and Transcriptomic Analysis of the Impact of Androgen Stimulation and Darolutamide Inhibition

ncbi.nlm.nih.gov/pmc/articl...

Simple Summary

Abstract

Several inhibitors of androgen receptor (AR) function are approved for prostate cancer treatment, and their impact on gene transcription has been described. However, the ensuing effects at the protein level are far less well understood. We focused on the AR signaling inhibitor darolutamide and confirmed its strong AR binding and antagonistic activity using the high throughput cellular thermal shift assay (CETSA HT). Then, we generated comprehensive, quantitative proteomic data from the androgen-sensitive prostate cancer cell line VCaP and compared them to transcriptomic data. Following treatment with the synthetic androgen R1881 and darolutamide, global mass spectrometry-based proteomics and label-free quantification were performed. We found a generally good agreement between proteomic and transcriptomic data upon androgen stimulation and darolutamide inhibition. Similar effects were found both for the detected expressed genes and their protein products as well as for the corresponding biological programs. However, in a few instances there was a discrepancy in the magnitude of changes induced on gene expression levels compared to the corresponding protein levels, indicating post-transcriptional regulation of protein abundance. Chromatin immunoprecipitation DNA sequencing (ChIP-seq) and Hi-C chromatin immunoprecipitation (HiChIP) revealed the presence of androgen-activated AR-binding regions and long-distance AR-mediated loops at these genes.

3.3. Androgen Stimulation and Darolutamide Inhibition Induce Largely Concordant Changes at the Gene and Protein Levels

We then focused on genes and proteins differentially expressed in cells treated by the androgen and darolutamide combination, compared to androgen treatment alone. Remarkable effects on known androgen-regulated genes and other androgen-dependent pathways, such as cholesterol homeostasis, fatty acid metabolism, unfolded protein response and cell cycle, were observed upon R1881 plus darolutamide application (Figure 3C). Scatter plot analysis showed a strong negative correlation between changes induced by R1881 compared to R1881 plus darolutamide, both in RNA (Figure S2A) and protein (Figure S2B) levels, demonstrating that darolutamide potently reverted expression changes induced by R1881. While hundreds of genes were found up- or down-regulated two-fold and more on the transcriptional level following treatment with R1881 and darolutamide, compared to R1881 alone, the magnitude of change was lower in the protein levels, with fewer proteins passing the two-fold threshold (Table S1).

Go to:

4. Discussion

………,

Androgen treatment strongly induced gene expression programs related to cell proliferation and survival, both at the transcription and protein expression levels.

The stimulatory effects we observed following androgen application were potently reverted by additional darolutamide treatment, in line with the high anti-proliferative activity of this compound in androgen-dependent prostate cancer models and its efficacy in the clinic [24,25,49]……..,

[Ramp7]

I am no expert on the exact mechanisms involved. But I do have a general understanding. I just recently cycled Darolutamide for a couple of months. Now back to a high T state. As Denmeade said to me, we may be able to turn this into a chronic condition.

[KocoPr in reply to Ramp7]

I am also cycling betwen prpoianate and daro. Presently on daro. I did notice that the daro cycle is not effecting me as when i was on it full time. I think the high T resets my overall hormones.

Question, do you still take a continuous AR antagonist? Im thinking of getting off the Orgovyx.

[PCaWarrior in reply to KocoPr]

If you get off the orgovyx, you should closely monitor your testosterone on the low phases. If it recovers you are not doing BAT (mine recovered from suppression in three months). If your body's T is 300 ng/dl then you are cycling from 2500+ to 300. High T to normal T. This is one of the most dangerous hormonal environments.

Caveat: If you are CRPC and trying to resensitize this might work. The data we have is for conventional BAT so you'd be striking out on your own. I think that for resensitization a long 1000-1800 ng/dl might be best. Not proven and I would try conventional cypionate BAT first.

[KocoPr in reply to PCaWarrior]

Got ya. I haven’t done it, but do get lazy on taking Orgo on high T except near end of high T. Stay religious with the t during daro cycle though.

[PCaWarrior in reply to KocoPr]

I'm going to skip my next Lupron shot and monitor T. I wasn't planning on doing that but my insurance conked out.

I hate insurance companies. And I hate that our approval process is so slow. It should be left up to the MO to decide what therapy is best for you. Not the insurance companies and twenty year behind the times FDA.

[KocoPr in reply to PCaWarrior]

I never heard lupron being denied!it is the first line SOC

[PCaWarrior in reply to KocoPr]

Insurance used to pay almost all of it. Last time it cost me over $600. Yearly cap maybe. I didn't check into it. Easier just to monitor my T for a cycle. Should be fine.

[lancer82801 in reply to PCaWarrior]

I read this exchange with interest. I would solicit your comments or anyone else, for that matter. I have been using estradiol patches exclusively and continuously for ADT for the past 3.5 years (0.1mg/day). Recently, I have been injecting Testosterone cypionate(800mg) when my PSA reaches its low point 0.04, Testosterone 9.7 It takes about 4 months to then return the PSA to 0.04 or 0.05 and testosterone to <10. Continuously using estradiol patches (2 twice weekly). Clearly it takes longer for the Estradiol to suppress the testosterone. I have been wondering if I could not reinject the testosterone when the blood testosterone reached about 150 as Dr Denmeade is doing, instead of waiting so long. My Doctor is hesitant to try it as we do see a bounce in PSA up to 0.64 or so each time. this bounce is expected with what I read about the saturation model for testosterone replacement (link attached) PSA has returned to the 0.04 range each time though

Lancer

research

[KocoPr in reply to lancer82801]

There is more than one way to keep the beast fed but caged and your doing a great job of it.

[PCaWarrior in reply to lancer82801]

If you use propionate instead of cypionate you would extend the high T cycle a lot. Seven times shorter half-life.

Or you could use Androgel. Androgel has an even shorter half-life but is more expensive, kind of a hassle to use (have to use it twice a day), and needs to be dosed high to get the T levels we need.

Or, what you could do, is use cypionate and then start adding some Androgel towards the end of the cypionate cycle. If you can't get propionate (empower pharmacy in Houston compounds it) I would do the cyp/androgel mix.

There is some research that shows that high estrogen at the same time as high t is not beneficial. Friedman says it is very dangerous. I use Lupron. The reasons to use high estrogen for ADT mostly go away when we use testosterone. And, if low E is an issue, we can replace a little bit during the low T phase. No need to do that on the high T phase.

BAT: cyp and androgel

[lancer82801 in reply to PCaWarrior]

Thanks for the response and thoughtful solution. Perchance, do you have the references where you cited the estrogen and testosterone being high is not beneficial ?

[PCaWarrior in reply to lancer82801]

The Estradiol-Dihydrotestosterone model of prostate cancer | Theoretical Biology and Medical Modelling | Full Text

tbiomed.biomedcentral.com/a...

In mice:

• T of 2500 ng/dl and normal E (10-60 pg/ml) – PCa did not develop

• No T and no E - PCa did not develop

• T of 2500 ng/dl and E of 675 pg/ml – PCa developed

Estrogen-Initiated Transformation of Prostate Epithelium Derived from Normal Human Prostate Stem-Progenitor Cells – PMC

ncbi.nlm.nih.gov/pmc/articl...

No high quality evidence so I talked to Edward Friedman (not surprising that there isn't much research on this - not many people are injecting testosterone and using estrogen at the same time).

[PCaWarrior in reply to lancer82801]

Neither here nor there in realistic applications but the PSA rise is expected and is because androgens overexpress KLK3 (thus PSA). If we look at DHT for example (you might see R1881 in cell studies - for various reasons R1881 is used as a DHT proxy by many) KLK3 expression continues to rise as DHT increases.

I performed this experiment on myself. I took T to 30,000 ng/dl. PSA went crazy. I had intermediate PSA points around T=1000, 2000, 3000, 4000.

By the way, do not do this experiment on yourself. It took a few months for my PSA to start acting reasonably. Very weird stuff. Two mets exploded in growth (stable per RECIST but I don't think they specify timeframes) and my ctDNA went to zero. A good news/bad news scenario.

Edit: The increase is not linear. As I go from 500 to 1500 as far as I can tell it seems somewhat linear. Above that the slope decreases (PSA vs T).

[lancer82801 in reply to PCaWarrior]

Thank you! Good information !! I would think, high estrogens in the intact male would be generating a lot of interest, as that is at least part of the treatment administered to the transgender community. Of course I say that, while some Doctors think it is okay to operated these poor young men and women with no clear idea of how profound the changes they are causing will affect them going forward!!

My sample of one does not seem to be following in the expected (from these papers) direction. Of course there is the fact that the prostate is no longer present in my case. so I am dealing with the escaped cells, survivors and offspring only. I have no evidence that it has metastasized from the prostatic bed. i do think that it is prudent to not try testosterone back to back as in the BAT model. Just the one injection seems to work pretty well (at least for the 5 times I have used it)

[PCaWarrior in reply to lancer82801]

Do you inject 800 mg of testosterone cypionate? Have you measured your serum levels at the max?

That is a hefty dose of testosterone. 400-500 mg is typically used.

[lancer82801 in reply to PCaWarrior]

Yes and yes

Prior to injection

PSA 0.04 Testosterone 9.72 and DHT <50 Estradiol 310

Inject 800mg Testosterone cypionate

28 days later

Testosterone 773 and PSA 0.39 Estradiol 260 DHT 518 within the next 30 days the PSA further increased to 0.64 and Testosterone decreased to 151 and DHT was 133 while Estradiol was 146

I have monthly tests of all parameters and thus 80 days later

Testosterone 14.80, PSA 0.06, Estradiol 134.97 and DHT <50

Some of the times the testosterone spikes well above 1000 but has the same time and duration back to 10 or less (90 days or so)

My doc calls it "astounding" and does not want to mess with it. Can't blame him but after reading this attached paper I had wondered about a follow up dose of Testosterone before it descended below about 150

Any thoughts?

research

[PCaWarrior in reply to lancer82801]

Ok. I was wondering if you had measured the peak T a day or so after the injection.

So I'm clear, you inject the 800 mg, don't do anything else, and 28 days following the only injection your T is 773 ng/dl.

And you're on ADT the entire time? (high estrogen in your case)

A day or so after injection you should get up to around 4500 ng/dl.

Your MO says it's working so I wouldn't want to arbitrarily change it. Are the reasons you want to change it QoL related?

We don't have any clinical trial data or much experimental data for that program (you are the first person that I know of).

[lancer82801 in reply to PCaWarrior]

No I have not measured the peak just days after injection. My supposition is that since the T is down to 10 or less it just takes more T to bring it back up. And, the goal was to obtain Supraphysiologic levels of T Like the BAT

That is correct I have not stopped the ADT (Estradiol patches ) for over two years. with no side effects that I am aware other than those associated with low T

I am totally satisfied with this treatment compared to the Lupron type ADT's this is sooo much easier on me in terms of side effects.

I wanted to add more testosterone for QoL enhancements but truthfully it probably wont make much difference as my age and circumstances leave me not many options.

My Doctor said prior to Lupron he treated many patients with estrogen in the form of DES ( Diethylstilbestrol ) I brought the Patch trial to him (use of Estradiol patches as the ADT) and he said sure lets try it.

your are no doubt aware of the VA retrospective study which found an increased incidence of cancer in patients treated with oral DES as the ADT. No doubt, Just a coincidence that Lupron got FDA clearance about the same time. The theory that it is the route of administration (oral and thus to the liver first for degredation) and the synthertic nature of DES that caused the problems with other cancers being stimulated.

What changes would you suggest?

[PCaWarrior in reply to lancer82801]

First, you are doing well and there are many reasons why BAT is preferential to ADT. I'm over 5 years post diagnosis with a low PSA, HSPC, zero ctDNA. These were not the predictions for me. Individual variance perhaps; however, my MO feels that my therapies are working (for over 2 years the only therapy is a modified BAT program).

DES causes an increase in CVD. There are reasons for this and most of the increase was in men who had existing cardiac issues. Transdermal bypasses first pass liver metabolism and has no increased CVD risk per PATCH and other studies. It is a far better stand-alone therapy for ADT than Lupron or other GnRH drugs (e.g., Firmagon and orgovyx).

What I would suggest is to think about the 800 mg of cypionate that you are using. Might be ok. We don't have data for it. That's quite high. I'd think about lowering it to 400-500 mg. You'll go low T faster and you'll match up with the main BAT trials. Your T will still go quite high using 400-500 mg of cypionate.

If possible, I think a consultation with Denmeade would be helpful ([e-mail]). Maybe talk to your MO about this?

Since QoL isn't a concern, I wouldn't mess with adding propionate or Androgel to what is working.

[lancer82801 in reply to PCaWarrior]

Thanks a lot and glad you are doing well

I still wonder how long I can go with the high levels of estrogen I am maintaining

I did use 600 mg testosterone one time and it did not get me above 1000 so went back to 800. I will talk to my MO and we could easily cut back Your comments are much appreciated

[PCaWarrior in reply to lancer82801]

I'm guessing you didn't time it for Cmax (maximum concentration). It's easy to miss the max. For cypionate it usually occurs 1-3 days after injection.

A rough estimate is max total testosterone = 5 * T-cyp mg.

400 reliably gets me over 2000 ng/dl. My lean mass is fairly high. You'd need to have more muscle mass than Big Rami (former Mr. Olympia title holder) for 600 mg of T-cyp to only get to 1000 ng/dl.

You or your MO can look into testosterone levels. I found this picture that shows what 250 mg of enanthate does. Cyp is similar to enanthate. (Ignore the injectable undecanoate. It has a very long half-life and isn't close to cyp. )

If you change formulas from cypionate to propionate or use Androgel or something else, the multiplier changes. It can be calculated from half-life and dose and delivery mechanism. Fortunately for us it is very simple to calculate the serum levels obtained from injectable testosterones.

serum t

[lancer82801 in reply to PCaWarrior]

more good information Thanks I will take this to my MO and we can easily change the dose

[PCaWarrior in reply to lancer82801]

I want to add that Dr. Denmeade uses 400-500 mg of cypionate in his BAT trials. He reports that the obtained levels are > 1500 ng/dl. The clinical trials might be of interest to your MO. I would look at RESTORE, TRANSFORMER, and the in-progress STEP-UP.

[lancer82801 in reply to PCaWarrior]

Again I thank you for your willingness to share your knowledge.

[MateoBeach in reply to lancer82801]

Hi Lancer. You got very good information from PCaWarrior. I would add the following: It seems preferable in theory to maintain SPA (T > 1000ng/dL continuously during your high-T BAT cycles of whatever duration you choose, whether one month, two months or 3 months (as I do). Getting a high spike then having it decay over several weeks does not accomplish that. So the 800mg one-time T-cyp dose does not make sense to me. I tried 400 mg T-cyp every 2 weeks and found that even that did not maintain levels >1000 but dropped to below 350 by the two weeks. So now I use 250-300 mg T-Cyp every week. But switch to T-propionate 75mg every 2 days for the last 4 weeks of my 3-month high T BAT cycles. Then I do with one month of Orgovyx combined with half dose darolutamide for the first 2 weeks of that.

There is ZERO reason to use ADT during the high-T cycles! Any tiny remnant of testicular testosterone is absent or buried by the high external source. No reason to use continuous estrogen through those periods as an ADT. And not sure how much recovery there would be during my castrate month. I could probably drop the Orgovyx and just use the darolutamide.

Darolutamide is not yet available from India generic sellers yet. So sourcing is problematic. I get my T-propionate from Dragon Pharma. Paul

[lancer82801 in reply to MateoBeach]

Thank you for this information I now have a lot to look into.

Interesting I thought the goal was to cycle between supraphysiologic levels (>1000 T and lower <200T) Dr Denmeade's protocol , can you enlarge on your protocol?

What do you use to monitor your cycles? I presume it would be PSA ?

At the onset I had consulted with several different oncologist and have been met with outright refusal to even consult with me on my requests to use Estradiol. So the Doc i am using is amenable to helping, but he is a busy man and I am just one of many patients. I have provided him information throughout and he is most willing to look at it .

I am taking to heart PCaWarrior's caution about the high levels of Testosterone I have been using and will change that. I have considered reinjecting the Testosterone when the levels decrease to <200 as Dr Denmeade has done. My Doctor is hesitant to change because, what we are doing seems to be working. I need more evidence that repeating the testosterone at some level (you are doing it at 1000?)for several injections. I have linked below on the Saturation model for Testosterone, and I gave this to my Doctor. Is there any other literature I could show my Doc ? I am still unsure of what is the trigger that one needs to be monitoring ? I have not had any scans done since the first year after diagnosis and had no Lymph node or other distant metastasis (My PSA has been <1 for the last 3.5 years) Monitoring only with PSA, Estradiol, Testosterone, Free testosterone (DHT), and D-Dimer test monthly

Letter to the editor: Questioning the evidence behind the Saturation Model for testosterone replacement therapy in prostate cancericurology.org/DOIx.php?id=1...

I have wondered about stopping the estrogen but Dr Denmeade is pretty adamant about keeping up the ADT in his protocol and thus we have as well. I am anxious to learn how you manage your program.

I worry about the high Estrogen levels i am maintaining but to date have not recognized any problems. Judging by when I started on the high estrogen protocol it takes a while ( 4 patches twice weekly for a month) to effectively suppress the endogenous T and thus the PSA. Comparing just costs the estradiol patches ($90.00 every 6weeks) and intermittent testosterone injections are considerably less money than using even lupron as the ADT

thanks again for jumping in

I sincerely appreciate your willingness to provide your wisdom.

Lancer

[Scout4answers in reply to MateoBeach]

I could probably drop the Orgovyx and just use the darolutamide.

Are you saying you think you could just use a half dose of Daro for 1 month and get the same result? Or would it be a full dose of Daro?

Really like your idea of constant (weekly) shots of T-cyp to maintain T levels.

Would you explain why you " switch to T-propionate 75mg every 2 days for the last 4 weeks of my 3-month high T BAT cycles."

[MateoBeach in reply to Scout4answers]

1) Using Daro without Orgovyx for one month may be possible because testicular testosterone production may be near zero during that time. But I would have to test the theory by measuring testosterone during the last week of it. Easy to do.

2) T-cypionate has a half life of about 7 days, so it takes 5 weeks to clear to a castrate level. 2000 to 1000 to 500 to 250 to 125 to 62.5, that's 5 weeks, perhaps even 6 weeks to get to castrate! That is why 28 day BAT with one shot of T-cyp is suboptimal. Changing to T-propionate for the last several weeks has just a 24 hour half-life. It will be fully gone within a week after stopping it. And Darolutamide will block any that remains. If one was going to do BAT with just 4 weeks of high testosterone, then propionate would be a better choice for the whole program, that is what Russ favors and several are doing.

[KocoPr in reply to MateoBeach]

I have stopped Orgovyx for a test.

I will run. T panel from quest. Question i have is exactly when should i test the T panel?

Im thinking halfway through my 2week daro cycle with a T panel and end of daro cycle with just T.

Any ideas are appreciated.

[Scout4answers in reply to MateoBeach]

Thanks Paul

I am taking my last 3 month shot of Lupron on Dec 5 and then will see what T and PSA do after March 5 with no Lupron in my system, plan to take Abi and prednisone until March, then let my "vacation" start.

As of March '24 I will have had 30 months of Lupron but only 21 months post IMRT

My plan is to start a BAT program such as yours, if PSA rises above 0.20, it is currently 0.01.

Your thoughts are welcome .

[KocoPr in reply to Scout4answers]

0.01 is a good number. Iets hope you never rise to 0.2, but BAT is a great option. Do you plan to do Puals 3 month T plan?

Might I suggest you get darolutamide before you start BAT, also if you do start BAT might I also suggest you do a 2 week T-propianate to make sure PSA doesn’t get out of control. I myself in your shoes would definitely want to see a PSA increase but under 2.0 or 4.0 (your arbitrary oh crap factor) if your starting PSA is 0.2. You want a response but not a hyper response.

Optimal rise would be under 1.0 for a 2 week t-prop.

[Ramp7 in reply to KocoPr]

Yes, I am on Lupron (3 month injections)

[PCaWarrior]

Thanks for posting. The article shows that adding darolutamide at the same time as high DHT (our androgen pulses) inhibits some of the gene changes induced by DHT. The article is a study of proteomics and gene expression.

It supports the ARSI effects of darolutamide.

"R1881 plus darolutamide samples were closer to DMSO than R1881-treated samples were, indicating the reversal of the stimulatory effect of androgen by darolutamide. "

"darolutamide potently reverted expression changes induced by R1881"

But we do not do that. SPT and darolutamide are separated. ExBAT is a trial that is somewhat close to what we do. Denmeade said that propionate and darolutamide should work.

Day 1: Testosterone cypionate 400 mg IM on day 1.

Day 29-56: Darolutamide 600 mg twice a day orally for 4 weeks.

Day 57-63: Washout period.

exbat

[Ramp7]

Interesting, I am right at that door step, Oct. 10 (400 mg Cypionate) Day 29 (Nov. 8) I see MO Nov. 7.

[Farmhand]

Anyone here get Darolutamide through India Mart? I am CRPC metastatic (Although not currently) And so do not qualify for Daro.

Have been using BAT protocol variations for the past year and would like to use darolutamide instead of Enzalutamide Because of less severe side effects and faster washout time.

Not sure which company to order from… Anybody know some of the more reputable companies?

[Nfler in reply to Farmhand]

All day chemists out of India is who I have used n seem to have great results using their product…

[KocoPr in reply to Nfler]

That’s good to know

[Farmhand in reply to KocoPr]

Went to their site but could not find Darolutamide

[Nfler in reply to Farmhand]

Oh man ok, only suggestion is to check their reviews when u do so find some

[Scout4answers in reply to Nfler]

I am confused Nfler. I order drugs from All day Chemist in India and have never been able to order Darolutimide. Are you saying you have bought it there or are happy with the other products you buy there?

[Nfler in reply to Scout4answers]

Sorry at the confusion, yes just happy about other products obtained there…

[Farmhand in reply to Nfler]

Thanks!

[Ramp7]

If you find a cheaper compounder, please mention back here.