Australian high school students have done "a little Breaking Bad" bysynthesizing and effectively open-sourcing the drug famously hiked 5,000 percent in price by "pharma bro" Martin Shkreli. The drug they recreated, Daraprim, is used to treat infection caused by malaria and HIV and without it, many patients would die. "Working on a real-world problem definitely made us more enthusiastic," said 17-year-old Sydney Grammar student Austin Zhang. "The background to this [drug] made it seem more important."
Daraprim is a relatively simple compound and typically costs $12.99 AUD ($10) for fifty tablets in Australia. However, Shkreli's company, Turing Pharmaceuticals, has the exclusive rights to distribute the specific Daraprim formulation in the USA (it's known as Pyrimethamine elsewhere), even though the drug was developed in 1953, and is long out of patent.
To get a new version approved, a company would have to compare it Turing's FDA-approved product with their permission, which isn't likely -- the company limits sales to doctors and pharmacies, making it difficult to reverse-engineer. Pharma companies would therefore need to go through an onerous approval process that probably wouldn't be worth it, considering that less than 10,000 Daraprim prescriptions are written in the US per year. (The US uses a "closed distribution" system which differs from most other countries.)
Though the open source Daraprim literally debunks Shkreli's premise that the drug is "underpriced" (supply and demand aside), it probably won't directly help anyone. Shkreli himself dismissed the work with a tweet, saying, "how is that showing anyone up? Almost any drug can be made at small scale for a low price. Glad it makes u feel good tho [sic]."
However, that doesn't mean that the exercise was useless. In fact, the students didn't just follow a recipe, they actually reverse-engineered the drug, checking their progress using spectral analysis on each new compound.
They also posted the work on Github, letting experts from the Open Source Malaria Consortium (OSM) (endorsed by Bill Gates) provide some help. For instance, the process used to manufacture Daraprim would be dangerous for students to replicate in a small high school lab. "They had to change things as some reagents were nasty and dangerous so some invention was needed on their part," said Todd.
After achieving a "beautiful" spectrograph, they finished with 3.7 grams of pure pyrimethamine, worth about $110,000 on the US market, and presented the results at a prestigious symposium. The OSM also posted aguide for making the drug that could help anyone else who wanted to try. That's quite an accomplishment for 16- and 17-year-old students, even if they can't actually sell it. And they sort of proved that as tempting as it is to hate Shkreli, he's merely profiting from a US system that's much friendlier to pharmaceutical companies than other countries.
Gene editing could provide a cure for HIV
While antiretroviral drugs do a good job of keeping HIV infections under control, scientists are working hard to come up with a full cure for the condition. A team of researchers from the Lewis Katz School of Medicine at Temple University is making real progress in that regard, successfully testing a gene editing system, demonstrating its ability to eliminate the virus from DNA in human cells grown in culture.
Since it was first discovered in the 1980s, HIV and AIDS has caused in excess of 25 million deaths. Antiretroviral drugs are effective at controlling the infection, but if patients stop having the treatment, the infection quickly takes hold and health deteriorates rapidly.
Finding a cure is problematic, as it's difficult to eradicate the virus once it's integrated into CD4+ T-cells – the primary cells infected by HIV. Some recent attempts to find a cure have focused on actually reactivating the virus to prompt a strong immune response to kill it, but no study has yet yielded positive results.
The Temple University team's approach is more subtle, using CRISPR/Cas9 gene editing technology to target and remove the virus from DNA. The tech is made up of a guide RNA, which is used to locates the HIV-1 virus in the DNA, and a nuclease, which is then able to edit it out of the sequence. Once the virus is eliminated, the cell's own mechanisms step in and tie up the loose ends of the genome.
Working with T-cells from patients infected with HIV-1, grown in culture, the researchers were able to demonstrate the technology removing the virus, and continuing to protect against further infection following the treatment.
The researchers also looked at whether the treatment had any off-target effects, or caused any toxicity – an essential investigation should the technology ever be widely used. Analyzing cells following the treatment, the team used ultra-deep whole-genome sequencing to determine that no off-target effects had occurred, including any potential changes to the gene expression of the cell. Observations of the cells showed them to be growing and functioning in a healthy manner.
The results provide the most complete set of data on the pioneering new treatment, and while widespread use is still a long way off, the researchers can now move ahead with the technology, working towards a full cure for HIV.
Full details of the study were published online in the journal Scientific Reports.
New Findings Provide a Design for an HIV Vaccine Germline-Targeting Immunogen
Some people infected with HIV naturally produce antibodies that effectively neutralize many strains of the rapidly mutating virus, and scientists are working to develop a vaccine capable of inducing such “broadly neutralizing” antibodies that can prevent HIV infection.
An emerging vaccine strategy involves immunizing people with a series of different engineered HIV proteins as immunogens to teach the immune system to produce broadly neutralizing antibodies against HIV. This strategy depends on the ability of the first immunogen to bind and activate special cells, known as broadly neutralizing antibody precursor B cells, which have the potential to develop into broadly neutralizing antibody-producing B cells.
A research team has now found that the right precursor (“germline”) cells for one kind of HIV broadly neutralizing antibody are present in most people, and has described the design of an HIV vaccine germline-targeting immunogen capable of binding those B cells. The findings by scientists from The Scripps Research Institute (TSRI), the International AIDS Vaccine Initiative (IAVI) and the La Jolla Institute for Allergy and Immunology were published in Science.
“We found that almost everybody has these broadly neutralizing antibody precursors, and that a precisely engineered protein can bind to these cells that have potential to develop into HIV broadly neutralizing antibody-producing cells, even in the presence of competition from other immune cells,” said the study’s lead author, William Schief, TSRI professor and director, Vaccine Design of the IAVI Neutralizing Antibody Center at TSRI, in whose lab the engineered HIV vaccine protein was developed.
The body’s immune system contains a large pool of different precursor B cells so it can respond to a wide variety of pathogens. But that also means that precursor B cells able to recognize a specific feature on a virus surface are exceedingly rare within the total pool of B cells.
“The challenge for vaccine developers is to determine if an immunogen can present a particular viral surface in a way that distinct B cells can be activated, proliferate and be useful,” said study co-author Shane Crotty, professor at the La Jolla Institute. “Using a new technique, we were able to show—well in advance of clinical trials—that most humans actually have the right B cells that will bind to this vaccine candidate. It is remarkable that protein design can be so specific as to ’find’ one in a million cells, demonstrating the feasibility of this new vaccine strategy.”
The work offers encouraging insights for a planned Phase 1 clinical trial to test a nanoparticle version of the engineered HIV vaccine protein, the “eOD-GT8 60mer.” “The goal of the clinical study will be to test safety and the ability of this engineered protein to elicit the desired immune response in humans that would look like the start of broadly neutralizing antibody development,” Schief said. “Data from this new study was also important for designing the clinical trial, including the size and the methods of analysis.”
In June, scientists from TSRI, IAVI and The Rockefeller University reported that the eOD-GT8 60mer produced antibody responses in mice that showed some of the traits necessary to recognize and inhibit HIV. If the eOD-GT8 60mer performs similarly in humans, additional boost immunogens are thought to be needed to ultimately induce broadly neutralizing antibodies that can block HIV.
The new work also provides a method for researchers to assess whether other new vaccine proteins can bind their intended precursor B cells. This method is a valuable tool in the design of more targeted and effective vaccines against AIDS, providing the ability to vet germline-targeting immunogens before testing them in large, time-consuming and costly clinical trials.
Looking at blood donated by healthy volunteers, the scientists found B cells that were capable of creating “VRC01-class” antibodies that recognized a critical surface patch, or epitope, of HIV. VRC01-class broadly neutralizing antibodies are a group of antibodies isolated from different individuals that appear to have developed in a very similar way, and it has been hypothesized that the starting VRC01-class B cells were very similar in the different people. The eOD-GT8 60mer is designed to engage these precursor B cells to initiate HIV broadly neutralizing antibody development.
Other contributors to the paper, “HIV-1 broadly neutralizing antibody precursor B cells revealed by germline-targeting immunogen,” included Joseph Jardine, Daniel Kulp, Colin Havenar-Daughton, Anita Sarkar, Bryan Briney, Devin Sok, Fabian Sesterhenn, June Ereno-Orbea, Oleksandr Kalyuzhniy, Isaiah Deresa, Xiaozhen Hu, Skye Spencer, Meaghan Jones, Erik Georgeson, Jumiko Adachi, Michael Kubitz, Allan decamp, Jean-Philippe Julien, Ian Wilson and Dennis Burton. This work was supported by the International AIDS Vaccine Initiative Neutralizing Antibody Consortium and Center; the Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology and Harvard; the Bayer Science and Education Foundation; the Helen Hay Whitney Foundation; Howard Hughes Medical Institute; Bill & Melinda Gates Foundation; and the National Institute of Allergy and Infectious Diseases (P01 AI094419, Center for HIV/AIDS Vaccine Immunology & Immunogen Discovery (CHAVI-ID) 1UM1AI100663, P01 AI82362 and R01 AI084817.)